TY  - CONF
AU  - Lukić, Ivana
AU  - Dragačević, Luka
AU  - Panić, Marko
AU  - Stamenković, Marina
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/878
AB  - In the post-COVID-19 era, there has been a significant increase in the development of mRNA vaccines not only against various diseases besides SARS-CoV-2, but also to treat cancer and genetic disorders. These vaccines, revolutionizing vaccinology, offer rapid pandemic response, high efficacy, minimal side effects, and cost-effectiveness. Achieving these benefits hinges on seamlessly integrating mRNA production steps, from plasmid DNA cloning to lipid nanoparticle formulation. This overview aims to comprehend or circumvent pitfalls in plasmid DNA cloning, a critical initial step in mRNA vaccine production. The focus is on achieving accurate insert sequence and gene expression, and it highlights the critical role of plasmid DNA design in ensuring vaccine effectiveness. Our research project entitled “Role of macroautophagy in lipid nanoparticle mRNA delivery and adjuvanticity” recognized the significance of this aspect. During our research, we designed a plasmid DNA cloning vector to incorporate the GFP-SARS-CoV-2 Spike gene. The vector was carefully constructed with several key features, including a high-copy plasmid, pUC18/pUC19 vector backbone with a robust T7 promoter, origin of replication, multiple cloning sites, polyadenylation signal, and ampicillin resistance for bacterial selection. Despite careful design, challenges like poly-A tail deletion may arise, prompting the exploration of stable large-size and low-copy vectors, as well as linear and bacteriophage vectors. But, for largescale production and regulatory compliance, vector systems must be scalable and well-documented. Commercial vectors and automated synthesis facilitate gene construction, with artificial intelligence ensuring sequence accuracy. Precision is crucial for complex antigens, as seen in tuberculosis mRNA vaccine development. Addressing these challenges demands a combining of molecular biology techniques, computational tools, and collaboration with experts in microbiology, molecular biology, and vaccine development. The design’s scalability and documentation are vital for large-scale production and regulatory compliance, emphasizing the multifaceted approach required for successful mRNA vaccine development.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design
EP  - 157
SP  - 157
UR  - https://hdl.handle.net/21.15107/rcub_intor_878
ER  - 

@conference{
author = "Lukić, Ivana and Dragačević, Luka and Panić, Marko and Stamenković, Marina and Kojić, Milan",
year = "2024",
abstract = "In the post-COVID-19 era, there has been a significant increase in the development of mRNA vaccines not only against various diseases besides SARS-CoV-2, but also to treat cancer and genetic disorders. These vaccines, revolutionizing vaccinology, offer rapid pandemic response, high efficacy, minimal side effects, and cost-effectiveness. Achieving these benefits hinges on seamlessly integrating mRNA production steps, from plasmid DNA cloning to lipid nanoparticle formulation. This overview aims to comprehend or circumvent pitfalls in plasmid DNA cloning, a critical initial step in mRNA vaccine production. The focus is on achieving accurate insert sequence and gene expression, and it highlights the critical role of plasmid DNA design in ensuring vaccine effectiveness. Our research project entitled “Role of macroautophagy in lipid nanoparticle mRNA delivery and adjuvanticity” recognized the significance of this aspect. During our research, we designed a plasmid DNA cloning vector to incorporate the GFP-SARS-CoV-2 Spike gene. The vector was carefully constructed with several key features, including a high-copy plasmid, pUC18/pUC19 vector backbone with a robust T7 promoter, origin of replication, multiple cloning sites, polyadenylation signal, and ampicillin resistance for bacterial selection. Despite careful design, challenges like poly-A tail deletion may arise, prompting the exploration of stable large-size and low-copy vectors, as well as linear and bacteriophage vectors. But, for largescale production and regulatory compliance, vector systems must be scalable and well-documented. Commercial vectors and automated synthesis facilitate gene construction, with artificial intelligence ensuring sequence accuracy. Precision is crucial for complex antigens, as seen in tuberculosis mRNA vaccine development. Addressing these challenges demands a combining of molecular biology techniques, computational tools, and collaboration with experts in microbiology, molecular biology, and vaccine development. The design’s scalability and documentation are vital for large-scale production and regulatory compliance, emphasizing the multifaceted approach required for successful mRNA vaccine development.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design",
pages = "157-157",
url = "https://hdl.handle.net/21.15107/rcub_intor_878"
}

Lukić, I., Dragačević, L., Panić, M., Stamenković, M.,& Kojić, M.. (2024). mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 157-157.
https://hdl.handle.net/21.15107/rcub_intor_878

Lukić I, Dragačević L, Panić M, Stamenković M, Kojić M. mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:157-157.
https://hdl.handle.net/21.15107/rcub_intor_878 .

Lukić, Ivana, Dragačević, Luka, Panić, Marko, Stamenković, Marina, Kojić, Milan, "mRNA vaccine manufacturing – challenges in plasmid DNA cloning vector design" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):157-157,
https://hdl.handle.net/21.15107/rcub_intor_878 .

TY  - CONF
AU  - Tsibulskaya, Darya
AU  - Blagojević, Veljko
AU  - Terzić-Vidojević, Amarela
AU  - Lukić, Ivana
AU  - Vasić, Marko
AU  - Dragačević, Luka
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/875
AB  - Autoaggregation, the ability to self-aggregate, is widespread among both Gram-positive and Gram-negative bacteria. The functional role of aggregation is not fully understood, but it is believed to be involved in the adaptation of bacteria to environmental conditions (PMID: 31294207). One interesting class of compounds responsible for the aggregation of lactic acid bacteria is aggregation factors—surface high-molecular-weight proteins rich in threonine and lysine (PMID: 30027759). Recently, our research group discovered a new strain of Streptococcus thermophilus in the mountainous regions of Serbia, exhibiting an aggregation phenotype. Aggregation phenotype was confirmed visually and using microscopy. Complete genome of Agg+ strain was sequenced using NGS and a gene encoding a potential aggregation factor, which was named aggS was identified. The predicted threonine (12.5%) and lysine (10.5%) rich protein contains 2367 amino acids, with an average molecular weight of 255986.63 Da. AggS also contains two cysteine residues, whereas previously well-described aggregation factors of this type did not contain any cysteine residues. The predicted protein includes an N-terminal YSIRK-like signal sequence and an LPXTG cell wall anchor domain. It has 6 Mucin binding domain repeats alternating with 6 Mub B2-like domain repeats. Additionally, we found a region resembling an ice-binding domain. Given that these bacteria endure prolonged periods of low temperatures, it can be speculated that this surface membrane protein also helps the bacteria withstand freezing. The fact that the alignment using BLASTp revealed AggS to be most closely related to an uncharacterised protein from the genome of Lactococcus garvieae, along with the discovery of a transposase gene sequence upstream of the gene, suggests that the aggregation factor was likely acquired through horizontal gene transfer. We plan to clone it into a shuttle vector and investigate the aggregation phenotype using a heterologous expression system in Lactococcus lactis, as well as explore its other functions.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - Description of a new potential aggregation factor from the Streptococcus thermophilus genome
EP  - 110
SP  - 110
UR  - https://hdl.handle.net/21.15107/rcub_intor_875
ER  - 

@conference{
author = "Tsibulskaya, Darya and Blagojević, Veljko and Terzić-Vidojević, Amarela and Lukić, Ivana and Vasić, Marko and Dragačević, Luka and Kojić, Milan",
year = "2024",
abstract = "Autoaggregation, the ability to self-aggregate, is widespread among both Gram-positive and Gram-negative bacteria. The functional role of aggregation is not fully understood, but it is believed to be involved in the adaptation of bacteria to environmental conditions (PMID: 31294207). One interesting class of compounds responsible for the aggregation of lactic acid bacteria is aggregation factors—surface high-molecular-weight proteins rich in threonine and lysine (PMID: 30027759). Recently, our research group discovered a new strain of Streptococcus thermophilus in the mountainous regions of Serbia, exhibiting an aggregation phenotype. Aggregation phenotype was confirmed visually and using microscopy. Complete genome of Agg+ strain was sequenced using NGS and a gene encoding a potential aggregation factor, which was named aggS was identified. The predicted threonine (12.5%) and lysine (10.5%) rich protein contains 2367 amino acids, with an average molecular weight of 255986.63 Da. AggS also contains two cysteine residues, whereas previously well-described aggregation factors of this type did not contain any cysteine residues. The predicted protein includes an N-terminal YSIRK-like signal sequence and an LPXTG cell wall anchor domain. It has 6 Mucin binding domain repeats alternating with 6 Mub B2-like domain repeats. Additionally, we found a region resembling an ice-binding domain. Given that these bacteria endure prolonged periods of low temperatures, it can be speculated that this surface membrane protein also helps the bacteria withstand freezing. The fact that the alignment using BLASTp revealed AggS to be most closely related to an uncharacterised protein from the genome of Lactococcus garvieae, along with the discovery of a transposase gene sequence upstream of the gene, suggests that the aggregation factor was likely acquired through horizontal gene transfer. We plan to clone it into a shuttle vector and investigate the aggregation phenotype using a heterologous expression system in Lactococcus lactis, as well as explore its other functions.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "Description of a new potential aggregation factor from the Streptococcus thermophilus genome",
pages = "110-110",
url = "https://hdl.handle.net/21.15107/rcub_intor_875"
}

Tsibulskaya, D., Blagojević, V., Terzić-Vidojević, A., Lukić, I., Vasić, M., Dragačević, L.,& Kojić, M.. (2024). Description of a new potential aggregation factor from the Streptococcus thermophilus genome. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 110-110.
https://hdl.handle.net/21.15107/rcub_intor_875

Tsibulskaya D, Blagojević V, Terzić-Vidojević A, Lukić I, Vasić M, Dragačević L, Kojić M. Description of a new potential aggregation factor from the Streptococcus thermophilus genome. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:110-110.
https://hdl.handle.net/21.15107/rcub_intor_875 .

Tsibulskaya, Darya, Blagojević, Veljko, Terzić-Vidojević, Amarela, Lukić, Ivana, Vasić, Marko, Dragačević, Luka, Kojić, Milan, "Description of a new potential aggregation factor from the Streptococcus thermophilus genome" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):110-110,
https://hdl.handle.net/21.15107/rcub_intor_875 .

TY  - CONF
AU  - Prijić, Ivana
AU  - Panić, Marko
AU  - Simić, Mihajlo
AU  - Blagojević, Veljko
AU  - Ćuruvija, Ivana
AU  - Lukić, Ivana
AU  - Dragačević, Luka
AU  - Kojić, Milan
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/877
AB  - Diphtheria toxin is a single polypeptide chain produced by toxigenic strains of Corynebacterium diphtheriae that causes the disease diphtheria in humans by gaining entry into the cytoplasm of cells and inhibiting protein synthesis. Formaldehyde (chemical) detoxification converts diphtheria toxin into toxoid, which is used in diphtheria vaccine production. Recombinant, genetically detoxified diphtheria toxin is superior in terms of safety and purity, but it has still not found its application in recombinant diphtheria vaccine production. Both chemically and genetically inactivated forms of the diphtheria toxin have proven effective as protein carriers in conjugate vaccines. The goal of this study was to create a plasmid construct which can be used to express a genetically inactivated diphtheria toxin. Gene coding for diphtheria toxin was cloned into pMALHisEk expression vector and introduced into DH5α competent Escherichia coli cells. Three site-directed point mutations, which led to three amino acid substitutions (G52E-substitutes glycine with glutamic acid, G79D- substitutes glycine with aspartic acid, E148D- substitutes glutamic acid with aspartic acid) were conducted. A single G52E amino acid substitution is responsible for the loss of the enzymatic activity of the diphtheria toxin. G79D is recognized as a good candidate site for combining with other mutations in vaccine development and E148D may be a good candidate as carrier protein because it could reduce both the stability of NAD binding and catalytic activity of the enzyme. Each individual mutation is sufficient for toxin inactivation, but together they ensure non-toxicity, preventing reversion to the wild-type sequence. All mutations were confirmed by DNA sequencing. Recombinant diphtheria toxoid could serve as a potential vaccine epitope or protein carrier for conjugate vaccines. Further optimization of recombinant protein expression in Escherichia coli should provide sufficient quantities of soluble recombinant protein for further testing of its safety, immunogenicity and protection.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - Inactivation of diphtheria toxin by site-directed mutagenesis
EP  - 115
SP  - 115
UR  - https://hdl.handle.net/21.15107/rcub_intor_877
ER  - 

@conference{
author = "Prijić, Ivana and Panić, Marko and Simić, Mihajlo and Blagojević, Veljko and Ćuruvija, Ivana and Lukić, Ivana and Dragačević, Luka and Kojić, Milan",
year = "2024",
abstract = "Diphtheria toxin is a single polypeptide chain produced by toxigenic strains of Corynebacterium diphtheriae that causes the disease diphtheria in humans by gaining entry into the cytoplasm of cells and inhibiting protein synthesis. Formaldehyde (chemical) detoxification converts diphtheria toxin into toxoid, which is used in diphtheria vaccine production. Recombinant, genetically detoxified diphtheria toxin is superior in terms of safety and purity, but it has still not found its application in recombinant diphtheria vaccine production. Both chemically and genetically inactivated forms of the diphtheria toxin have proven effective as protein carriers in conjugate vaccines. The goal of this study was to create a plasmid construct which can be used to express a genetically inactivated diphtheria toxin. Gene coding for diphtheria toxin was cloned into pMALHisEk expression vector and introduced into DH5α competent Escherichia coli cells. Three site-directed point mutations, which led to three amino acid substitutions (G52E-substitutes glycine with glutamic acid, G79D- substitutes glycine with aspartic acid, E148D- substitutes glutamic acid with aspartic acid) were conducted. A single G52E amino acid substitution is responsible for the loss of the enzymatic activity of the diphtheria toxin. G79D is recognized as a good candidate site for combining with other mutations in vaccine development and E148D may be a good candidate as carrier protein because it could reduce both the stability of NAD binding and catalytic activity of the enzyme. Each individual mutation is sufficient for toxin inactivation, but together they ensure non-toxicity, preventing reversion to the wild-type sequence. All mutations were confirmed by DNA sequencing. Recombinant diphtheria toxoid could serve as a potential vaccine epitope or protein carrier for conjugate vaccines. Further optimization of recombinant protein expression in Escherichia coli should provide sufficient quantities of soluble recombinant protein for further testing of its safety, immunogenicity and protection.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "Inactivation of diphtheria toxin by site-directed mutagenesis",
pages = "115-115",
url = "https://hdl.handle.net/21.15107/rcub_intor_877"
}

Prijić, I., Panić, M., Simić, M., Blagojević, V., Ćuruvija, I., Lukić, I., Dragačević, L.,& Kojić, M.. (2024). Inactivation of diphtheria toxin by site-directed mutagenesis. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 115-115.
https://hdl.handle.net/21.15107/rcub_intor_877

Prijić I, Panić M, Simić M, Blagojević V, Ćuruvija I, Lukić I, Dragačević L, Kojić M. Inactivation of diphtheria toxin by site-directed mutagenesis. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:115-115.
https://hdl.handle.net/21.15107/rcub_intor_877 .

Prijić, Ivana, Panić, Marko, Simić, Mihajlo, Blagojević, Veljko, Ćuruvija, Ivana, Lukić, Ivana, Dragačević, Luka, Kojić, Milan, "Inactivation of diphtheria toxin by site-directed mutagenesis" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):115-115,
https://hdl.handle.net/21.15107/rcub_intor_877 .

TY  - CONF
AU  - Lukić, Ivana
AU  - Popović, Mina
AU  - Miljković, Radmila
AU  - Tsibulskaya, Darya
AU  - Panić, Marko
AU  - Dragačević, Luka
AU  - Stojanović, Marijana
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/874
AB  - Limosilactobacillus reuteri demonstrates a significant
role in treating gastrointestinal diseases
through the synthesis of various health-promoting
factors. These include mucus-binding proteins,
reactive oxygen species-scavenging enzymes, antimicrobial
agents (reuterin is capable of inhibiting
the growth of a wide spectrum of microorganisms),
vitamins (folate and vitamin B12), and unique
exopolysaccharides. Different strains of L. reuteri
exhibit strain-specific anti-inflammatory effects,
influencing the expression of immune-related
factors such as IL-10 and TNF-α (PMID: 20798357;
PMID: 22207578). Furthermore, the mitigating
impact of L. reuteri strains on inflammation is confirmed
in vivo and in vitro with the implication of
an interaction between probiotics and immune
cells in the intestinal mucosa (PMID: 22207578).
Our study aimed to investigate the potential anti-
inflammatory effects of daily treatment with autochthonous
probiotic strain L. reuteri B2 (PMID:
33932415) could have an anti-inflammatory effect
on local immune response. In a 14-day experiment
with Intor Swiss: Albino mice (n=10), those treated
with L. reuteri B2 (5x106 CFU/mL, 100 μl) showed a
favorable impact on the gut’s inflammatory environment.
Histological analyses of colon samples
and intraperitoneal macrophages revealed lower
myeloperoxidase (MPO) activity, reduced production
of superoxide ions, IFNγ, IL-6, and TNFα, along
with an enhanced production of IL-10 in L. reuteri
B2 treated mice compared to untreated ones. Notably,
histopathological preparations did not show
significant differences between the groups. The
study suggests that L. reuteri B2 may be valuable
for further evaluation in managing, preventing,
and treating inflammatory bowel diseases. The
presented findings contribute to understanding
the specific anti-inflammatory effects of this strain
on the local immune response, supporting its potential
as a therapeutic agent.
PB  - Serbian Society for Microbiology
C3  - XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
T1  - The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration
EP  - 38
SP  - 38
UR  - https://hdl.handle.net/21.15107/rcub_intor_874
ER  - 

@conference{
author = "Lukić, Ivana and Popović, Mina and Miljković, Radmila and Tsibulskaya, Darya and Panić, Marko and Dragačević, Luka and Stojanović, Marijana",
year = "2024",
abstract = "Limosilactobacillus reuteri demonstrates a significant
role in treating gastrointestinal diseases
through the synthesis of various health-promoting
factors. These include mucus-binding proteins,
reactive oxygen species-scavenging enzymes, antimicrobial
agents (reuterin is capable of inhibiting
the growth of a wide spectrum of microorganisms),
vitamins (folate and vitamin B12), and unique
exopolysaccharides. Different strains of L. reuteri
exhibit strain-specific anti-inflammatory effects,
influencing the expression of immune-related
factors such as IL-10 and TNF-α (PMID: 20798357;
PMID: 22207578). Furthermore, the mitigating
impact of L. reuteri strains on inflammation is confirmed
in vivo and in vitro with the implication of
an interaction between probiotics and immune
cells in the intestinal mucosa (PMID: 22207578).
Our study aimed to investigate the potential anti-
inflammatory effects of daily treatment with autochthonous
probiotic strain L. reuteri B2 (PMID:
33932415) could have an anti-inflammatory effect
on local immune response. In a 14-day experiment
with Intor Swiss: Albino mice (n=10), those treated
with L. reuteri B2 (5x106 CFU/mL, 100 μl) showed a
favorable impact on the gut’s inflammatory environment.
Histological analyses of colon samples
and intraperitoneal macrophages revealed lower
myeloperoxidase (MPO) activity, reduced production
of superoxide ions, IFNγ, IL-6, and TNFα, along
with an enhanced production of IL-10 in L. reuteri
B2 treated mice compared to untreated ones. Notably,
histopathological preparations did not show
significant differences between the groups. The
study suggests that L. reuteri B2 may be valuable
for further evaluation in managing, preventing,
and treating inflammatory bowel diseases. The
presented findings contribute to understanding
the specific anti-inflammatory effects of this strain
on the local immune response, supporting its potential
as a therapeutic agent.",
publisher = "Serbian Society for Microbiology",
journal = "XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april",
title = "The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration",
pages = "38-38",
url = "https://hdl.handle.net/21.15107/rcub_intor_874"
}

Lukić, I., Popović, M., Miljković, R., Tsibulskaya, D., Panić, M., Dragačević, L.,& Stojanović, M.. (2024). The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april
Serbian Society for Microbiology., 38-38.
https://hdl.handle.net/21.15107/rcub_intor_874

Lukić I, Popović M, Miljković R, Tsibulskaya D, Panić M, Dragačević L, Stojanović M. The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration. in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april. 2024;:38-38.
https://hdl.handle.net/21.15107/rcub_intor_874 .

Lukić, Ivana, Popović, Mina, Miljković, Radmila, Tsibulskaya, Darya, Panić, Marko, Dragačević, Luka, Stojanović, Marijana, "The anti-inflammatory effect of Limosilactobacillus Reuteri b2 administration" in XIII Congress of microbiologists of Serbia with international participation, Mikromed regio 5, From biotechnology to human and planetary health, 4-6 april (2024):38-38,
https://hdl.handle.net/21.15107/rcub_intor_874 .

TY  - JOUR
AU  - Virijević, Katarina
AU  - Živanović, Marko N.
AU  - Nikolić, Dalibor
AU  - Milivojević, Nevena
AU  - Pavić, Jelena
AU  - Morić, Ivana
AU  - Šenerović, Lidija
AU  - Dragačević, Luka
AU  - Thurner, Philipp J.
AU  - Rufin, Manuel
AU  - Andriotis, Orestis G.
AU  - Ljujić, Biljana
AU  - Miletić Kovačević, Marina
AU  - Papić, Miloš
AU  - Filipović, Nenad
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/872
AB  - Here, an artificial intelligence (AI)-based approach was employed to optimize the production of electrospun scaffolds for in vivo wound healing applications. By combining polycaprolactone (PCL) and poly(ethylene glycol) (PEG) in various concentration ratios, dissolved in chloroform (CHCl3) and dimethylformamide (DMF), 125 different polymer combinations were created. From these polymer combinations, electrospun nanofiber meshes were produced and characterized structurally and mechanically via microscopic techniques, including chemical composition and fiber diameter determination. Subsequently, these data were used to train a neural network, creating an AI model to predict the optimal scaffold production solution. Guided by the predictions and experimental outcomes of the AI model, the most promising scaffold for further in vitro analyses was identified. Moreover, we enriched this selected polymer combination by incorporating antibiotics, aiming to develop electrospun nanofiber scaffolds tailored for in vivo wound healing applications. Our study underscores three noteworthy conclusions: (i) the application of AI is pivotal in the fields of material and biomedical sciences, (ii) our methodology provides an effective blueprint for the initial screening of biomedical materials, and (iii) electrospun PCL/PEG antibiotic-bearing scaffolds exhibit outstanding results in promoting neoangiogenesis and facilitating in vivo wound treatment.
PB  - American Chemical Society
T2  - ACS Applied Materials & Interfaces
T2  - ACS Applied Materials & InterfacesACS Appl. Mater. Interfaces
T1  - AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing
DO  - 10.1021/acsami.4c03266
ER  - 

@article{
author = "Virijević, Katarina and Živanović, Marko N. and Nikolić, Dalibor and Milivojević, Nevena and Pavić, Jelena and Morić, Ivana and Šenerović, Lidija and Dragačević, Luka and Thurner, Philipp J. and Rufin, Manuel and Andriotis, Orestis G. and Ljujić, Biljana and Miletić Kovačević, Marina and Papić, Miloš and Filipović, Nenad",
year = "2024",
abstract = "Here, an artificial intelligence (AI)-based approach was employed to optimize the production of electrospun scaffolds for in vivo wound healing applications. By combining polycaprolactone (PCL) and poly(ethylene glycol) (PEG) in various concentration ratios, dissolved in chloroform (CHCl3) and dimethylformamide (DMF), 125 different polymer combinations were created. From these polymer combinations, electrospun nanofiber meshes were produced and characterized structurally and mechanically via microscopic techniques, including chemical composition and fiber diameter determination. Subsequently, these data were used to train a neural network, creating an AI model to predict the optimal scaffold production solution. Guided by the predictions and experimental outcomes of the AI model, the most promising scaffold for further in vitro analyses was identified. Moreover, we enriched this selected polymer combination by incorporating antibiotics, aiming to develop electrospun nanofiber scaffolds tailored for in vivo wound healing applications. Our study underscores three noteworthy conclusions: (i) the application of AI is pivotal in the fields of material and biomedical sciences, (ii) our methodology provides an effective blueprint for the initial screening of biomedical materials, and (iii) electrospun PCL/PEG antibiotic-bearing scaffolds exhibit outstanding results in promoting neoangiogenesis and facilitating in vivo wound treatment.",
publisher = "American Chemical Society",
journal = "ACS Applied Materials & Interfaces, ACS Applied Materials & InterfacesACS Appl. Mater. Interfaces",
title = "AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing",
doi = "10.1021/acsami.4c03266"
}

Virijević, K., Živanović, M. N., Nikolić, D., Milivojević, N., Pavić, J., Morić, I., Šenerović, L., Dragačević, L., Thurner, P. J., Rufin, M., Andriotis, O. G., Ljujić, B., Miletić Kovačević, M., Papić, M.,& Filipović, N.. (2024). AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing. in ACS Applied Materials & Interfaces
American Chemical Society..
https://doi.org/10.1021/acsami.4c03266

Virijević K, Živanović MN, Nikolić D, Milivojević N, Pavić J, Morić I, Šenerović L, Dragačević L, Thurner PJ, Rufin M, Andriotis OG, Ljujić B, Miletić Kovačević M, Papić M, Filipović N. AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing. in ACS Applied Materials & Interfaces. 2024;.
doi:10.1021/acsami.4c03266 .

Virijević, Katarina, Živanović, Marko N., Nikolić, Dalibor, Milivojević, Nevena, Pavić, Jelena, Morić, Ivana, Šenerović, Lidija, Dragačević, Luka, Thurner, Philipp J., Rufin, Manuel, Andriotis, Orestis G., Ljujić, Biljana, Miletić Kovačević, Marina, Papić, Miloš, Filipović, Nenad, "AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing" in ACS Applied Materials & Interfaces (2024),
https://doi.org/10.1021/acsami.4c03266 . .

TY  - DATA
AU  - Virijević, Katarina
AU  - Živanović, Marko N.
AU  - Nikolić, Dalibor
AU  - Milivojević, Nevena
AU  - Pavić, Jelena
AU  - Morić, Ivana
AU  - Šenerović, Lidija
AU  - Dragačević, Luka
AU  - Thurner, Philipp J.
AU  - Rufin, Manuel
AU  - Andriotis, Orestis G.
AU  - Ljujić, Biljana
AU  - Miletić Kovačević, Marina
AU  - Papić, Miloš
AU  - Filipović, Nenad
PY  - 2024
UR  - http://intor.torlakinstitut.com/handle/123456789/873
AB  - Figure S1. Publication Trends in “Electrospinning”, “Electrospinning + PCL + PEG”, “Electrospinning + Wound Healing” and “Electrospinning + Artificial Intelligence + Neural Network” Research (2001-2022) Table S1. Synonyms and Related Terms for Electrospinning in Research (2001-2022) Table S2. Input data structured for ANN – CSV data file Figure S2. Basic visualization of the dependence of output data (vertical axis) on individual input data (horizontal axis). Figure S3. Schematic representation of the neural network Figure S4. Graph of RMSE relation between training data set (blue line) and validation data set (orange line) depending on the number of neurons in the hidden layer Figure S5. Visual representation of ANN precision; the horizontal axis represents the percent of the real result, and the vertical axis represents the percent of ANN prediction; the training set (blue dots), prediction set (orange dots) Scheme S1. Electrospinning-Ready Polymer and Solvent Combinations. “Substance” is PCL or PCL combined with PEG. The substance is dissolved in mass concentrations from 17 to 28% in CHCl3 or a combination of CHCl3 and DMF. Figure S6. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 1: PCL in CHCl3. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S7. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 2: PCL in CHCl3:DMF=1:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S8. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 3: PCL in CHCl3:DMF=1:3. A) 17% B) 18% C) 19% Figure S9. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 4: PCL in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% Figure S10. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 5: PCL:PEG=1:1 in CHCl3. A) 17% B) 18% C) 20% D) 21% E) 22% F) 24% G) 25% H) 26% I) 27% J) 28% Figure S11. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 6: PCL:PEG=1:1 in CHCl3:DMF=1:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S12. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 7: PCL:PEG=1:1 in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% Figure S13. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 8: PCL:PEG=3:1 in CHCl3:DMF=1:1. A) 17% B) 19% C) 20% D) 21% E) 22% F) 24% G) 26% Figure S14. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 9: PCL:PEG=3:1 in CHCl3:DMF=1:3. A) 17% B) 18% C) 19% D) 22% Figure S15. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 10: PCL:PEG=3:1 in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% Figure S16. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 15: PCL:PEG=1:3 in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% Figure S17. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 16: PCL:PEG=7:3 in CHCl3:DMF=7:3. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S18. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 17: PCL:PEG=3:1 in CHCl3. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% Figure S19. The action of scaffolds bearing antibiotics on selected strains of bacteria by disk diffusion method. Figure S20. Chick Embryo CAM Assay Procedure: A) Egg selection B) Egg disinfection with 10% of iodine solution C) Inoculation and preparation for scaffold insertion D) Egg’s incubation E) Daily monitoring of embryo development and possible contamination F) Sacrifice of treated embryos and fixation with 4% PFA G) CAM membrane preparation H) Image capture and evaluation of blood vessels
PB  - American Chemical Society
T2  - ACS Applied Materials & Interfaces
T1  - Supplementary information for the article: Virijević, K.; Živanović, M. N.; Nikolić, D.; Milivojević, N.; Pavić, J.; Morić, I.; Šenerović, L.; Dragačević, L.; Thurner, P. J.; Rufin, M.; Andriotis, O. G.; Ljujić, B.; Miletić Kovačević, M.; Papić, M.; Filipović, N. AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing. ACS Appl. Mater. Interfaces 2024. https://doi.org/10.1021/acsami.4c03266.
DO  - doi.org/10.1021/acsami.4c03266
ER  - 

@misc{
author = "Virijević, Katarina and Živanović, Marko N. and Nikolić, Dalibor and Milivojević, Nevena and Pavić, Jelena and Morić, Ivana and Šenerović, Lidija and Dragačević, Luka and Thurner, Philipp J. and Rufin, Manuel and Andriotis, Orestis G. and Ljujić, Biljana and Miletić Kovačević, Marina and Papić, Miloš and Filipović, Nenad",
year = "2024",
abstract = "Figure S1. Publication Trends in “Electrospinning”, “Electrospinning + PCL + PEG”, “Electrospinning + Wound Healing” and “Electrospinning + Artificial Intelligence + Neural Network” Research (2001-2022) Table S1. Synonyms and Related Terms for Electrospinning in Research (2001-2022) Table S2. Input data structured for ANN – CSV data file Figure S2. Basic visualization of the dependence of output data (vertical axis) on individual input data (horizontal axis). Figure S3. Schematic representation of the neural network Figure S4. Graph of RMSE relation between training data set (blue line) and validation data set (orange line) depending on the number of neurons in the hidden layer Figure S5. Visual representation of ANN precision; the horizontal axis represents the percent of the real result, and the vertical axis represents the percent of ANN prediction; the training set (blue dots), prediction set (orange dots) Scheme S1. Electrospinning-Ready Polymer and Solvent Combinations. “Substance” is PCL or PCL combined with PEG. The substance is dissolved in mass concentrations from 17 to 28% in CHCl3 or a combination of CHCl3 and DMF. Figure S6. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 1: PCL in CHCl3. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S7. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 2: PCL in CHCl3:DMF=1:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S8. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 3: PCL in CHCl3:DMF=1:3. A) 17% B) 18% C) 19% Figure S9. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 4: PCL in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% Figure S10. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 5: PCL:PEG=1:1 in CHCl3. A) 17% B) 18% C) 20% D) 21% E) 22% F) 24% G) 25% H) 26% I) 27% J) 28% Figure S11. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 6: PCL:PEG=1:1 in CHCl3:DMF=1:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S12. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 7: PCL:PEG=1:1 in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% Figure S13. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 8: PCL:PEG=3:1 in CHCl3:DMF=1:1. A) 17% B) 19% C) 20% D) 21% E) 22% F) 24% G) 26% Figure S14. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 9: PCL:PEG=3:1 in CHCl3:DMF=1:3. A) 17% B) 18% C) 19% D) 22% Figure S15. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 10: PCL:PEG=3:1 in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% Figure S16. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 15: PCL:PEG=1:3 in CHCl3:DMF=3:1. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% Figure S17. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 16: PCL:PEG=7:3 in CHCl3:DMF=7:3. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% G) 23% H) 24% I) 25% J) 26% K) 27% Figure S18. Fiber Diameter Distribution Analysis of Electrospun-Derived Scaffold for Series 17: PCL:PEG=3:1 in CHCl3. A) 17% B) 18% C) 19% D) 20% E) 21% F) 22% Figure S19. The action of scaffolds bearing antibiotics on selected strains of bacteria by disk diffusion method. Figure S20. Chick Embryo CAM Assay Procedure: A) Egg selection B) Egg disinfection with 10% of iodine solution C) Inoculation and preparation for scaffold insertion D) Egg’s incubation E) Daily monitoring of embryo development and possible contamination F) Sacrifice of treated embryos and fixation with 4% PFA G) CAM membrane preparation H) Image capture and evaluation of blood vessels",
publisher = "American Chemical Society",
journal = "ACS Applied Materials & Interfaces",
title = "Supplementary information for the article: Virijević, K.; Živanović, M. N.; Nikolić, D.; Milivojević, N.; Pavić, J.; Morić, I.; Šenerović, L.; Dragačević, L.; Thurner, P. J.; Rufin, M.; Andriotis, O. G.; Ljujić, B.; Miletić Kovačević, M.; Papić, M.; Filipović, N. AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing. ACS Appl. Mater. Interfaces 2024. https://doi.org/10.1021/acsami.4c03266.",
doi = "doi.org/10.1021/acsami.4c03266"
}

Virijević, K., Živanović, M. N., Nikolić, D., Milivojević, N., Pavić, J., Morić, I., Šenerović, L., Dragačević, L., Thurner, P. J., Rufin, M., Andriotis, O. G., Ljujić, B., Miletić Kovačević, M., Papić, M.,& Filipović, N.. (2024). Supplementary information for the article: Virijević, K.; Živanović, M. N.; Nikolić, D.; Milivojević, N.; Pavić, J.; Morić, I.; Šenerović, L.; Dragačević, L.; Thurner, P. J.; Rufin, M.; Andriotis, O. G.; Ljujić, B.; Miletić Kovačević, M.; Papić, M.; Filipović, N. AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing. ACS Appl. Mater. Interfaces 2024. https://doi.org/10.1021/acsami.4c03266.. in ACS Applied Materials & Interfaces
American Chemical Society..
https://doi.org/doi.org/10.1021/acsami.4c03266

Virijević K, Živanović MN, Nikolić D, Milivojević N, Pavić J, Morić I, Šenerović L, Dragačević L, Thurner PJ, Rufin M, Andriotis OG, Ljujić B, Miletić Kovačević M, Papić M, Filipović N. Supplementary information for the article: Virijević, K.; Živanović, M. N.; Nikolić, D.; Milivojević, N.; Pavić, J.; Morić, I.; Šenerović, L.; Dragačević, L.; Thurner, P. J.; Rufin, M.; Andriotis, O. G.; Ljujić, B.; Miletić Kovačević, M.; Papić, M.; Filipović, N. AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing. ACS Appl. Mater. Interfaces 2024. https://doi.org/10.1021/acsami.4c03266.. in ACS Applied Materials & Interfaces. 2024;.
doi:doi.org/10.1021/acsami.4c03266 .

Virijević, Katarina, Živanović, Marko N., Nikolić, Dalibor, Milivojević, Nevena, Pavić, Jelena, Morić, Ivana, Šenerović, Lidija, Dragačević, Luka, Thurner, Philipp J., Rufin, Manuel, Andriotis, Orestis G., Ljujić, Biljana, Miletić Kovačević, Marina, Papić, Miloš, Filipović, Nenad, "Supplementary information for the article: Virijević, K.; Živanović, M. N.; Nikolić, D.; Milivojević, N.; Pavić, J.; Morić, I.; Šenerović, L.; Dragačević, L.; Thurner, P. J.; Rufin, M.; Andriotis, O. G.; Ljujić, B.; Miletić Kovačević, M.; Papić, M.; Filipović, N. AI-Driven Optimization of PCL/PEG Electrospun Scaffolds for Enhanced In Vivo Wound Healing. ACS Appl. Mater. Interfaces 2024. https://doi.org/10.1021/acsami.4c03266." in ACS Applied Materials & Interfaces (2024),
https://doi.org/doi.org/10.1021/acsami.4c03266 . .

TY  - JOUR
AU  - Dragačević, Luka
AU  - Lopandić, Zorana
AU  - Gavrović-Jankulović, Marija
AU  - Živković, Irena
AU  - Blagojević, Veljko
AU  - Polović, Natalija
AU  - Minić, Rajna
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/621
AB  - The surface of microorganisms is covered with carbohydrates, which makes them unique, self-sustaining glycan probes. Lectins are able to bind to these probes, and this interaction can be exploited for selecting microorganisms or novel lectins. To examine lectin-microorganism interactions, we have previously developed an enzyme-linked lectin sorbent assay (ELLSA) with whole bacterial cells. To further test the validity of this methodology, here we compare it with flow cytometry. For this purpose, we used biotinylated recombinantly produced lectin from Musa acuminata (BanLec), this lectin’s recombinantly produced chimera with green fluorescent protein (BanLec-eGFP) and a lectin from Ricinus communis (RCA120), both biotinylated and FITC labeled. Parallel testing showed equivalent results for the two methods, in terms of the presence or absence of binding, with signal intensity yielding high Pearson correlation coefficient of 0.8 for BanLec and 0.95 for RCA120. The ELLSA method demonstrated multiple advantages, such as reliability and convenience for high-throughput analysis; it also required less lectin and yielded more consistent results. As such, ELLSA proved to be a useful tool for profiling microbial glycan structures or testing novel lectins.
PB  - Springer
T2  - Applied Biochemistry and Biotechnology
T1  - Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans
EP  - 2060
SP  - 2047
VL  - 194
DO  - 10.1007/s12010-021-03772-w
ER  - 

@article{
author = "Dragačević, Luka and Lopandić, Zorana and Gavrović-Jankulović, Marija and Živković, Irena and Blagojević, Veljko and Polović, Natalija and Minić, Rajna",
year = "2022",
abstract = "The surface of microorganisms is covered with carbohydrates, which makes them unique, self-sustaining glycan probes. Lectins are able to bind to these probes, and this interaction can be exploited for selecting microorganisms or novel lectins. To examine lectin-microorganism interactions, we have previously developed an enzyme-linked lectin sorbent assay (ELLSA) with whole bacterial cells. To further test the validity of this methodology, here we compare it with flow cytometry. For this purpose, we used biotinylated recombinantly produced lectin from Musa acuminata (BanLec), this lectin’s recombinantly produced chimera with green fluorescent protein (BanLec-eGFP) and a lectin from Ricinus communis (RCA120), both biotinylated and FITC labeled. Parallel testing showed equivalent results for the two methods, in terms of the presence or absence of binding, with signal intensity yielding high Pearson correlation coefficient of 0.8 for BanLec and 0.95 for RCA120. The ELLSA method demonstrated multiple advantages, such as reliability and convenience for high-throughput analysis; it also required less lectin and yielded more consistent results. As such, ELLSA proved to be a useful tool for profiling microbial glycan structures or testing novel lectins.",
publisher = "Springer",
journal = "Applied Biochemistry and Biotechnology",
title = "Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans",
pages = "2060-2047",
volume = "194",
doi = "10.1007/s12010-021-03772-w"
}

Dragačević, L., Lopandić, Z., Gavrović-Jankulović, M., Živković, I., Blagojević, V., Polović, N.,& Minić, R.. (2022). Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. in Applied Biochemistry and Biotechnology
Springer., 194, 2047-2060.
https://doi.org/10.1007/s12010-021-03772-w

Dragačević L, Lopandić Z, Gavrović-Jankulović M, Živković I, Blagojević V, Polović N, Minić R. Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. in Applied Biochemistry and Biotechnology. 2022;194:2047-2060.
doi:10.1007/s12010-021-03772-w .

Dragačević, Luka, Lopandić, Zorana, Gavrović-Jankulović, Marija, Živković, Irena, Blagojević, Veljko, Polović, Natalija, Minić, Rajna, "Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans" in Applied Biochemistry and Biotechnology, 194 (2022):2047-2060,
https://doi.org/10.1007/s12010-021-03772-w . .

TY  - DATA
AU  - Dragačević, Luka
AU  - Lopandić, Zorana
AU  - Gavrović-Jankulović, Marija
AU  - Živković, Irena
AU  - Blagojević, Veljko
AU  - Polović, Natalija
AU  - Minić, Rajna
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/642
AB  - SDS-PAGE showing: Lane 1 – purified banana lectin – BanLec; Lane 2 – chimera of banana lectin and enhanced green fluorescent protein BanLec-eGFP; Lane 3 – MW – molecular weight markers; Table 1. Repeatability of flow cytometric detection of different quantities of BL-eGFP binding to C. albicans; Table 2. Repeatability of ELLSA detection of different quantities of BanLec-B binding to C. albicans; Table 3. Repeatability of flow cytometric detection of different quantities of RCA120-FITC binding to L. casei DG; Table 4. Repeatability of ELLSA detection of different quantities of RCA120-B binding to L. casei DG; Table 5. Reproducibility of BanLec-B binding to 21 different microorganisms. Experiments were done in three different laboratories, with preparing new plates each time; Fig. 1 Determining linearity of ELLSA method, by measuring binding between C.albicans and different quantity of BanLec-B; Fig. 2 Determining linearity of flow cytometry, by measuring binding between C.albicans and different quantity of BanLec-eGFP; Fig. 3 Determining linearity of ELLSA method, by measuring binding between L.casei DG and different quantity of RCA120; Fig. 4 Determining linearity of flow cytometry, by measuring binding between L.casei DG and different quantity of RCA120.
PB  - Springer
T2  - Applied Biochemistry and Biotechnology
T1  - Supplementary information for the article: Dragačević, L.; Lopandić, Z.; Gavrović-Jankulović, M.; Živković, I.; Blagojević, V.; Polović, N.; Minić, R. Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. Applied Biochemistry and Biotechnology 2022, 194, 2047–2060. https://doi.org/10.1007/s12010-021-03772-w.
EP  - 2060
SP  - 2047
VL  - 194
UR  - https://hdl.handle.net/21.15107/rcub_intor_642
ER  - 

@misc{
author = "Dragačević, Luka and Lopandić, Zorana and Gavrović-Jankulović, Marija and Živković, Irena and Blagojević, Veljko and Polović, Natalija and Minić, Rajna",
year = "2022",
abstract = "SDS-PAGE showing: Lane 1 – purified banana lectin – BanLec; Lane 2 – chimera of banana lectin and enhanced green fluorescent protein BanLec-eGFP; Lane 3 – MW – molecular weight markers; Table 1. Repeatability of flow cytometric detection of different quantities of BL-eGFP binding to C. albicans; Table 2. Repeatability of ELLSA detection of different quantities of BanLec-B binding to C. albicans; Table 3. Repeatability of flow cytometric detection of different quantities of RCA120-FITC binding to L. casei DG; Table 4. Repeatability of ELLSA detection of different quantities of RCA120-B binding to L. casei DG; Table 5. Reproducibility of BanLec-B binding to 21 different microorganisms. Experiments were done in three different laboratories, with preparing new plates each time; Fig. 1 Determining linearity of ELLSA method, by measuring binding between C.albicans and different quantity of BanLec-B; Fig. 2 Determining linearity of flow cytometry, by measuring binding between C.albicans and different quantity of BanLec-eGFP; Fig. 3 Determining linearity of ELLSA method, by measuring binding between L.casei DG and different quantity of RCA120; Fig. 4 Determining linearity of flow cytometry, by measuring binding between L.casei DG and different quantity of RCA120.",
publisher = "Springer",
journal = "Applied Biochemistry and Biotechnology",
title = "Supplementary information for the article: Dragačević, L.; Lopandić, Z.; Gavrović-Jankulović, M.; Živković, I.; Blagojević, V.; Polović, N.; Minić, R. Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. Applied Biochemistry and Biotechnology 2022, 194, 2047–2060. https://doi.org/10.1007/s12010-021-03772-w.",
pages = "2060-2047",
volume = "194",
url = "https://hdl.handle.net/21.15107/rcub_intor_642"
}

Dragačević, L., Lopandić, Z., Gavrović-Jankulović, M., Živković, I., Blagojević, V., Polović, N.,& Minić, R.. (2022). Supplementary information for the article: Dragačević, L.; Lopandić, Z.; Gavrović-Jankulović, M.; Živković, I.; Blagojević, V.; Polović, N.; Minić, R. Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. Applied Biochemistry and Biotechnology 2022, 194, 2047–2060. https://doi.org/10.1007/s12010-021-03772-w.. in Applied Biochemistry and Biotechnology
Springer., 194, 2047-2060.
https://hdl.handle.net/21.15107/rcub_intor_642

Dragačević L, Lopandić Z, Gavrović-Jankulović M, Živković I, Blagojević V, Polović N, Minić R. Supplementary information for the article: Dragačević, L.; Lopandić, Z.; Gavrović-Jankulović, M.; Živković, I.; Blagojević, V.; Polović, N.; Minić, R. Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. Applied Biochemistry and Biotechnology 2022, 194, 2047–2060. https://doi.org/10.1007/s12010-021-03772-w.. in Applied Biochemistry and Biotechnology. 2022;194:2047-2060.
https://hdl.handle.net/21.15107/rcub_intor_642 .

Dragačević, Luka, Lopandić, Zorana, Gavrović-Jankulović, Marija, Živković, Irena, Blagojević, Veljko, Polović, Natalija, Minić, Rajna, "Supplementary information for the article: Dragačević, L.; Lopandić, Z.; Gavrović-Jankulović, M.; Živković, I.; Blagojević, V.; Polović, N.; Minić, R. Comparison of Enzyme-Linked Lectin Sorbent Assay and Flow Cytometry for Profiling Microbial Glycans. Applied Biochemistry and Biotechnology 2022, 194, 2047–2060. https://doi.org/10.1007/s12010-021-03772-w." in Applied Biochemistry and Biotechnology, 194 (2022):2047-2060,
https://hdl.handle.net/21.15107/rcub_intor_642 .

TY  - JOUR
AU  - Knežević, Sanja
AU  - Kosanović, Dejana
AU  - Dragačević, Luka
AU  - Živković, Irena
AU  - Ilić, Vesna
AU  - Hajduković, Ljiljana
AU  - Savić, Olivera
AU  - Minić, Rajna
PY  - 2022
UR  - http://rimi.imi.bg.ac.rs/handle/123456789/1255
UR  - http://intor.torlakinstitut.com/handle/123456789/650
AB  - S. pneumoniae is an important human pathogen which has a polysaccharide capsule with virulent properties. This work aims to estimate the titres of S. pneumoniae specific IgG and IgA isotypes, with respect to age and sex. An in-house whole bacterial cell ELISA was used for the determination of relative levels and endpoint titres of IgG subclasses and IgA1 subclass specific for S. pneumoniae serogroup 1, and to quantify specific IgG1 and IgG2 levels. Significantly lower anti-pneumococcus IgG1 titres were found in older individuals, which was more pronounced in men. Lower IgG2 titres were detected in men over 50 years of age, in comparison to women under 50 years of age. The levels of IgG3 and IgG4 did not differ between different sex and age groups. Lower IgA1 levels were detected in male individuals in both age groups in comparison to females under 50 years of age. The levels of IgG1 showed a moderate correlation with IgG4 in younger individuals of both sexes (r = 0.61 in men and 0.63 in women) which was not noted in the older age group. We highlight the deficiency in humoral immunity in older people, especially male and suggest immunization of this population with pneumococcal vaccines.
PB  - Elsevier
T2  - Comparative Immunology, Microbiology and Infectious Diseases
T1  - Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1
SP  - 101834
VL  - 87
DO  - 10.1016/j.cimid.2022.101834
ER  - 

@article{
author = "Knežević, Sanja and Kosanović, Dejana and Dragačević, Luka and Živković, Irena and Ilić, Vesna and Hajduković, Ljiljana and Savić, Olivera and Minić, Rajna",
year = "2022",
abstract = "S. pneumoniae is an important human pathogen which has a polysaccharide capsule with virulent properties. This work aims to estimate the titres of S. pneumoniae specific IgG and IgA isotypes, with respect to age and sex. An in-house whole bacterial cell ELISA was used for the determination of relative levels and endpoint titres of IgG subclasses and IgA1 subclass specific for S. pneumoniae serogroup 1, and to quantify specific IgG1 and IgG2 levels. Significantly lower anti-pneumococcus IgG1 titres were found in older individuals, which was more pronounced in men. Lower IgG2 titres were detected in men over 50 years of age, in comparison to women under 50 years of age. The levels of IgG3 and IgG4 did not differ between different sex and age groups. Lower IgA1 levels were detected in male individuals in both age groups in comparison to females under 50 years of age. The levels of IgG1 showed a moderate correlation with IgG4 in younger individuals of both sexes (r = 0.61 in men and 0.63 in women) which was not noted in the older age group. We highlight the deficiency in humoral immunity in older people, especially male and suggest immunization of this population with pneumococcal vaccines.",
publisher = "Elsevier",
journal = "Comparative Immunology, Microbiology and Infectious Diseases",
title = "Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1",
pages = "101834",
volume = "87",
doi = "10.1016/j.cimid.2022.101834"
}

Knežević, S., Kosanović, D., Dragačević, L., Živković, I., Ilić, V., Hajduković, L., Savić, O.,& Minić, R.. (2022). Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1. in Comparative Immunology, Microbiology and Infectious Diseases
Elsevier., 87, 101834.
https://doi.org/10.1016/j.cimid.2022.101834

Knežević S, Kosanović D, Dragačević L, Živković I, Ilić V, Hajduković L, Savić O, Minić R. Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1. in Comparative Immunology, Microbiology and Infectious Diseases. 2022;87:101834.
doi:10.1016/j.cimid.2022.101834 .

Knežević, Sanja, Kosanović, Dejana, Dragačević, Luka, Živković, Irena, Ilić, Vesna, Hajduković, Ljiljana, Savić, Olivera, Minić, Rajna, "Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1" in Comparative Immunology, Microbiology and Infectious Diseases, 87 (2022):101834,
https://doi.org/10.1016/j.cimid.2022.101834 . .

TY  - JOUR
AU  - Knežević, Sanja
AU  - Kosanović, Dejana
AU  - Dragačević, Luka
AU  - Živković, Irena
AU  - Ilić, Vesna
AU  - Hajduković, Ljiljana
AU  - Savić, Olivera
AU  - Minić, Rajna
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/651
AB  - S. pneumoniae is an important human pathogen which has a polysaccharide capsule with virulent properties. This work aims to estimate the titres of S. pneumoniae specific IgG and IgA isotypes, with respect to age and sex. An in-house whole bacterial cell ELISA was used for the determination of relative levels and endpoint titres of IgG subclasses and IgA1 subclass specific for S. pneumoniae serogroup 1, and to quantify specific IgG1 and IgG2 levels. Significantly lower anti-pneumococcus IgG1 titres were found in older individuals, which was more pronounced in men. Lower IgG2 titres were detected in men over 50 years of age, in comparison to women under 50 years of age. The levels of IgG3 and IgG4 did not differ between different sex and age groups. Lower IgA1 levels were detected in male individuals in both age groups in comparison to females under 50 years of age. The levels of IgG1 showed a moderate correlation with IgG4 in younger individuals of both sexes (r = 0.61 in men and 0.63 in women) which was not noted in the older age group. We highlight the deficiency in humoral immunity in older people, especially male and suggest immunization of this population with pneumococcal vaccines.
PB  - Elsevier
T2  - Comparative Immunology, Microbiology and Infectious Diseases
T1  - Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1
SP  - 101834
VL  - 87
DO  - 10.1016/j.cimid.2022.101834
ER  - 

@article{
author = "Knežević, Sanja and Kosanović, Dejana and Dragačević, Luka and Živković, Irena and Ilić, Vesna and Hajduković, Ljiljana and Savić, Olivera and Minić, Rajna",
year = "2022",
abstract = "S. pneumoniae is an important human pathogen which has a polysaccharide capsule with virulent properties. This work aims to estimate the titres of S. pneumoniae specific IgG and IgA isotypes, with respect to age and sex. An in-house whole bacterial cell ELISA was used for the determination of relative levels and endpoint titres of IgG subclasses and IgA1 subclass specific for S. pneumoniae serogroup 1, and to quantify specific IgG1 and IgG2 levels. Significantly lower anti-pneumococcus IgG1 titres were found in older individuals, which was more pronounced in men. Lower IgG2 titres were detected in men over 50 years of age, in comparison to women under 50 years of age. The levels of IgG3 and IgG4 did not differ between different sex and age groups. Lower IgA1 levels were detected in male individuals in both age groups in comparison to females under 50 years of age. The levels of IgG1 showed a moderate correlation with IgG4 in younger individuals of both sexes (r = 0.61 in men and 0.63 in women) which was not noted in the older age group. We highlight the deficiency in humoral immunity in older people, especially male and suggest immunization of this population with pneumococcal vaccines.",
publisher = "Elsevier",
journal = "Comparative Immunology, Microbiology and Infectious Diseases",
title = "Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1",
pages = "101834",
volume = "87",
doi = "10.1016/j.cimid.2022.101834"
}

Knežević, S., Kosanović, D., Dragačević, L., Živković, I., Ilić, V., Hajduković, L., Savić, O.,& Minić, R.. (2022). Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1. in Comparative Immunology, Microbiology and Infectious Diseases
Elsevier., 87, 101834.
https://doi.org/10.1016/j.cimid.2022.101834

Knežević S, Kosanović D, Dragačević L, Živković I, Ilić V, Hajduković L, Savić O, Minić R. Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1. in Comparative Immunology, Microbiology and Infectious Diseases. 2022;87:101834.
doi:10.1016/j.cimid.2022.101834 .

Knežević, Sanja, Kosanović, Dejana, Dragačević, Luka, Živković, Irena, Ilić, Vesna, Hajduković, Ljiljana, Savić, Olivera, Minić, Rajna, "Age and gender associated changes in immunoglobulin subclass levels specific to S. pneumoniae, serotype 1" in Comparative Immunology, Microbiology and Infectious Diseases, 87 (2022):101834,
https://doi.org/10.1016/j.cimid.2022.101834 . .

TY  - JOUR
AU  - Ćuruvija, Ivana
AU  - Blagojević, Veljko
AU  - Dragačević, Luka
AU  - Vujić, Vesna
AU  - Lukić, Ivana
AU  - Stanojević, Stanislava
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/693
AB  - BCG vaccination induces a memory-like response in innate immune cells known as
trained immunity. In this study, we investigated the modification of innate immune
cells by BCG vaccination in acute peritoneal inflammation. We induced peritonitis with
thioglycollate (TG) in young Albino Oxford male rats which were immunised s.c. with
a BCG vaccine (BCG group) or saline (control group) 7 days prior. Peritoneal cells were
examined for 7 days after TG injection by flow cytometry, and for NO production and
peroxidase activity. Prior in vivo BCG priming altered TG-elicited peritoneal lavage
cells as following: increased in vitro LPS and BCG stimulated NO production from total
cells compared to adherent cells (day 1); increased cell number (days 3 and 5);
increased percentage of inflammatory monocytes (SSCmidCD43lowCD11bmid) and
eosinophils (SSCHihiS48+CD43hi), and a higher level of surface CD11b expression on
CD163+ macrophages (day 5); increased in vitro LPS and BCG stimulated peroxidase
activity (days 5 and 7); and increased percentage of CD163+MHCII+ cells (day 7). On
day 7, cells from both experimental groups showed no production of NO in response to
in vitro stimulation. We conclude that BCG vaccination had a substantial effect on the
acute phase of sterile inflammation, which may lead to the later observed phenotypic
and functional changes that could be seen as accelerated resolution of inflammation
and possibly point to trained immune response.
PB  - Wiley
T1  - BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?
EP  - 127
IS  - S2
SP  - 127
VL  - 52
UR  - https://hdl.handle.net/21.15107/rcub_intor_693
ER  - 

@article{
author = "Ćuruvija, Ivana and Blagojević, Veljko and Dragačević, Luka and Vujić, Vesna and Lukić, Ivana and Stanojević, Stanislava",
year = "2022",
abstract = "BCG vaccination induces a memory-like response in innate immune cells known as
trained immunity. In this study, we investigated the modification of innate immune
cells by BCG vaccination in acute peritoneal inflammation. We induced peritonitis with
thioglycollate (TG) in young Albino Oxford male rats which were immunised s.c. with
a BCG vaccine (BCG group) or saline (control group) 7 days prior. Peritoneal cells were
examined for 7 days after TG injection by flow cytometry, and for NO production and
peroxidase activity. Prior in vivo BCG priming altered TG-elicited peritoneal lavage
cells as following: increased in vitro LPS and BCG stimulated NO production from total
cells compared to adherent cells (day 1); increased cell number (days 3 and 5);
increased percentage of inflammatory monocytes (SSCmidCD43lowCD11bmid) and
eosinophils (SSCHihiS48+CD43hi), and a higher level of surface CD11b expression on
CD163+ macrophages (day 5); increased in vitro LPS and BCG stimulated peroxidase
activity (days 5 and 7); and increased percentage of CD163+MHCII+ cells (day 7). On
day 7, cells from both experimental groups showed no production of NO in response to
in vitro stimulation. We conclude that BCG vaccination had a substantial effect on the
acute phase of sterile inflammation, which may lead to the later observed phenotypic
and functional changes that could be seen as accelerated resolution of inflammation
and possibly point to trained immune response.",
publisher = "Wiley",
title = "BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?",
pages = "127-127",
number = "S2",
volume = "52",
url = "https://hdl.handle.net/21.15107/rcub_intor_693"
}

Ćuruvija, I., Blagojević, V., Dragačević, L., Vujić, V., Lukić, I.,& Stanojević, S.. (2022). BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?. 
Wiley., 52(S2), 127-127.
https://hdl.handle.net/21.15107/rcub_intor_693

Ćuruvija I, Blagojević V, Dragačević L, Vujić V, Lukić I, Stanojević S. BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?. 2022;52(S2):127-127.
https://hdl.handle.net/21.15107/rcub_intor_693 .

Ćuruvija, Ivana, Blagojević, Veljko, Dragačević, Luka, Vujić, Vesna, Lukić, Ivana, Stanojević, Stanislava, "BCG vaccination induced alterations of thioglycollate-elicited peritoneal phagocytes: a case of trained immunity?", 52, no. S2 (2022):127-127,
https://hdl.handle.net/21.15107/rcub_intor_693 .

TY  - CONF
AU  - Blagojević, Veljko
AU  - Ćuruvija, Ivana
AU  - Dragačević, Luka
AU  - Vujić, Vesna
AU  - Lukić, Ivana
AU  - Stanojević, Stanislava
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/694
AB  - Inflammation is a redistribution of immune cells, providing a more efficient elimination of the inflammatory offense. However, it is not limited to local microenvironment. In this study, the interaction of the effect of BCG priming and peritoneal inflammation on the remote inflammatory milieu of infected lung was investigated. Young male AO rats infected with Mycoplasma spp. were s.c. injected with BCG (3x105 CFU) or saline, and 7 days later received an i.p. injection of 7ml of thioglycollate (TG) or saline. Up to 7 days after TG injection, a broncho-alveolar lavage (BAL) was performed, and cells were analysed for their surface marker expression and NO production. Infected rats had a high percentage of HIS48HiCD11bHi neutrophils. BCG priming didn’t alter BAL cells phenotype, while TG injection increased the proportion of MHCII+CD11blow activated alveolar macrophages (aAMFs) on day 7. However, the BCG+TG group showed significant changes – percentage of HIS48HiCD11bHi neutrophils decreased from day 3, the share of aAMFs increased from day 5 and the share of MHCII+CD11b-AMFs increased on days 3-5. However, the percentage of B220+FSClow B lymphocytes were increased from day 1. Production of NO from BAL fluid cells was low in all groups. We conclude that BCG vaccination likely increased the number of circulating B lymphocytes, while TG-induced peritoneal inflammation potentially prevented their entry into the peritoneal cavity, forcing them into permissive tissues, such as lungs.
PB  - Wiley
T1  - Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation
EP  - 55
IS  - S2
SP  - 55
VL  - 52
UR  - https://hdl.handle.net/21.15107/rcub_intor_694
ER  - 

@conference{
author = "Blagojević, Veljko and Ćuruvija, Ivana and Dragačević, Luka and Vujić, Vesna and Lukić, Ivana and Stanojević, Stanislava",
year = "2022",
abstract = "Inflammation is a redistribution of immune cells, providing a more efficient elimination of the inflammatory offense. However, it is not limited to local microenvironment. In this study, the interaction of the effect of BCG priming and peritoneal inflammation on the remote inflammatory milieu of infected lung was investigated. Young male AO rats infected with Mycoplasma spp. were s.c. injected with BCG (3x105 CFU) or saline, and 7 days later received an i.p. injection of 7ml of thioglycollate (TG) or saline. Up to 7 days after TG injection, a broncho-alveolar lavage (BAL) was performed, and cells were analysed for their surface marker expression and NO production. Infected rats had a high percentage of HIS48HiCD11bHi neutrophils. BCG priming didn’t alter BAL cells phenotype, while TG injection increased the proportion of MHCII+CD11blow activated alveolar macrophages (aAMFs) on day 7. However, the BCG+TG group showed significant changes – percentage of HIS48HiCD11bHi neutrophils decreased from day 3, the share of aAMFs increased from day 5 and the share of MHCII+CD11b-AMFs increased on days 3-5. However, the percentage of B220+FSClow B lymphocytes were increased from day 1. Production of NO from BAL fluid cells was low in all groups. We conclude that BCG vaccination likely increased the number of circulating B lymphocytes, while TG-induced peritoneal inflammation potentially prevented their entry into the peritoneal cavity, forcing them into permissive tissues, such as lungs.",
publisher = "Wiley",
title = "Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation",
pages = "55-55",
number = "S2",
volume = "52",
url = "https://hdl.handle.net/21.15107/rcub_intor_694"
}

Blagojević, V., Ćuruvija, I., Dragačević, L., Vujić, V., Lukić, I.,& Stanojević, S.. (2022). Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation. 
Wiley., 52(S2), 55-55.
https://hdl.handle.net/21.15107/rcub_intor_694

Blagojević V, Ćuruvija I, Dragačević L, Vujić V, Lukić I, Stanojević S. Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation. 2022;52(S2):55-55.
https://hdl.handle.net/21.15107/rcub_intor_694 .

Blagojević, Veljko, Ćuruvija, Ivana, Dragačević, Luka, Vujić, Vesna, Lukić, Ivana, Stanojević, Stanislava, "Modifying Mycoplasma-infected lung immune cells through an intriguing interplay of BCG priming and peritoneal inflammation", 52, no. S2 (2022):55-55,
https://hdl.handle.net/21.15107/rcub_intor_694 .

TY  - JOUR
AU  - Lopandić, Zorana
AU  - Dragačević, Luka
AU  - Kosanović, Dejana
AU  - Burazer, Lidija
AU  - Gavrović-Jankulović, Marija
AU  - Minić, Rajna
PY  - 2022
UR  - http://intor.torlakinstitut.com/handle/123456789/787
AB  - In vivo animal models can provide worthy information on various aspects of asthma mechanism and pathogenesis. The genetic predisposition and phenotype of mice may affect the immune response itself. Here we compare the early immune response to Der p 2 or HDM allergen extract upon injection and inhalation in BALB/c and C57BL/6 mice. Female C57BL/6 and BALB/c mice were immunized with Der p 2 allergen subcutaneously followed by inhalation of Der p 2 or HDM extract. After challenge, the mice were euthanized; blood, bronchoalveolar lavage (BAL), spleens and lungs were collected. Cells from BAL were identified by May-Grünwald Giemsa staining and lung leukocyte populations were analyzed by flow cytometry. Serum antibody levels of Der p 2 specific IgE, IgG, IgG1 and IgG2a were assessed by ELISA, and cytokine secretion (IL-4, IFN-γ and IL-10) was evaluated upon stimulation with Der p 2 or HDM extract. The Th2 immune response was confirmed by elevated allergen-specific immunoglobulin E (IgE) and the allergic reaction was evidenced by infiltration of eosinophils and/or neutrophils into BAL. We found that BALB/c mice were inefficient in integrating local with systemic immune response, evidenced by almost no IgG or IgE production upon one subcutaneous injection and subsequent inhalation of Der p 2 allergen; also, the bronchoalveolar lavage infiltrate in these mice consisted of neutrophil infiltration, unlike C57BL/6 mice in which eosinophilic infiltrate predominated. The differences between BALB/c and C57BL/6 mice strains could be exploited for generating different types of responses to the Der p 2 allergen.
PB  - Elsevier
T2  - Journal of Immunological Methods
T2  - Journal of Immunological MethodsJournal of Immunological Methods
T1  - Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols
SP  - 113382
VL  - 511
DO  - 10.1016/j.jim.2022.113382
ER  - 

@article{
author = "Lopandić, Zorana and Dragačević, Luka and Kosanović, Dejana and Burazer, Lidija and Gavrović-Jankulović, Marija and Minić, Rajna",
year = "2022",
abstract = "In vivo animal models can provide worthy information on various aspects of asthma mechanism and pathogenesis. The genetic predisposition and phenotype of mice may affect the immune response itself. Here we compare the early immune response to Der p 2 or HDM allergen extract upon injection and inhalation in BALB/c and C57BL/6 mice. Female C57BL/6 and BALB/c mice were immunized with Der p 2 allergen subcutaneously followed by inhalation of Der p 2 or HDM extract. After challenge, the mice were euthanized; blood, bronchoalveolar lavage (BAL), spleens and lungs were collected. Cells from BAL were identified by May-Grünwald Giemsa staining and lung leukocyte populations were analyzed by flow cytometry. Serum antibody levels of Der p 2 specific IgE, IgG, IgG1 and IgG2a were assessed by ELISA, and cytokine secretion (IL-4, IFN-γ and IL-10) was evaluated upon stimulation with Der p 2 or HDM extract. The Th2 immune response was confirmed by elevated allergen-specific immunoglobulin E (IgE) and the allergic reaction was evidenced by infiltration of eosinophils and/or neutrophils into BAL. We found that BALB/c mice were inefficient in integrating local with systemic immune response, evidenced by almost no IgG or IgE production upon one subcutaneous injection and subsequent inhalation of Der p 2 allergen; also, the bronchoalveolar lavage infiltrate in these mice consisted of neutrophil infiltration, unlike C57BL/6 mice in which eosinophilic infiltrate predominated. The differences between BALB/c and C57BL/6 mice strains could be exploited for generating different types of responses to the Der p 2 allergen.",
publisher = "Elsevier",
journal = "Journal of Immunological Methods, Journal of Immunological MethodsJournal of Immunological Methods",
title = "Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols",
pages = "113382",
volume = "511",
doi = "10.1016/j.jim.2022.113382"
}

Lopandić, Z., Dragačević, L., Kosanović, D., Burazer, L., Gavrović-Jankulović, M.,& Minić, R.. (2022). Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols. in Journal of Immunological Methods
Elsevier., 511, 113382.
https://doi.org/10.1016/j.jim.2022.113382

Lopandić Z, Dragačević L, Kosanović D, Burazer L, Gavrović-Jankulović M, Minić R. Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols. in Journal of Immunological Methods. 2022;511:113382.
doi:10.1016/j.jim.2022.113382 .

Lopandić, Zorana, Dragačević, Luka, Kosanović, Dejana, Burazer, Lidija, Gavrović-Jankulović, Marija, Minić, Rajna, "Differences in mouse strains determine the outcome of Der p 2 allergy induction protocols" in Journal of Immunological Methods, 511 (2022):113382,
https://doi.org/10.1016/j.jim.2022.113382 . .

TY  - JOUR
AU  - Lopandic, Zorana
AU  - Dragačević, Luka
AU  - Popovic, Dragan
AU  - Andjelkovic, Uros
AU  - Minić, Rajna
AU  - Gavrovic-Jankulovic, Marija
PY  - 2021
UR  - http://intor.torlakinstitut.com/handle/123456789/615
AB  - Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms
PB  - MDPI
T2  - Biomolecules
T1  - BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms
IS  - 2
SP  - 180
VL  - 11(2)
VL  - 11
DO  - 10.3390/biom11020180
ER  - 

@article{
author = "Lopandic, Zorana and Dragačević, Luka and Popovic, Dragan and Andjelkovic, Uros and Minić, Rajna and Gavrovic-Jankulovic, Marija",
year = "2021",
abstract = "Fluorescently labeled lectins are useful tools for in vivo and in vitro studies of the structure and function of tissues and various pathogens such as viruses, bacteria, and fungi. For the evaluation of high-mannose glycans present on various glycoproteins, a three-dimensional (3D) model of the chimera was designed from the crystal structures of recombinant banana lectin (BanLec, Protein Data Bank entry (PDB): 5EXG) and an enhanced green fluorescent protein (eGFP, PDB 4EUL) by applying molecular modeling and molecular mechanics and expressed in Escherichia coli. BanLec-eGFP, produced as a soluble cytosolic protein of about 42 kDa, revealed β-sheets (41%) as the predominant secondary structures, with the emission peak maximum detected at 509 nm (excitation wavelength 488 nm). More than 65% of the primary structure was confirmed by mass spectrometry. Competitive BanLec-eGFP binding to high mannose glycans of the influenza vaccine (Vaxigrip®) was shown in a fluorescence-linked lectin sorbent assay (FLLSA) with monosaccharides (mannose and glucose) and wild type BanLec and H84T BanLec mutant. BanLec-eGFP exhibited binding to mannose residues on different strains of Salmonella in flow cytometry, with especially pronounced binding to a Salmonella Typhi clinical isolate. BanLec-eGFP can be a useful tool for screening high-mannose glycosylation sites on different microorganisms",
publisher = "MDPI",
journal = "Biomolecules",
title = "BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms",
number = "2",
pages = "180",
volume = "11(2), 11",
doi = "10.3390/biom11020180"
}

Lopandic, Z., Dragačević, L., Popovic, D., Andjelkovic, U., Minić, R.,& Gavrovic-Jankulovic, M.. (2021). BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. in Biomolecules
MDPI., 11(2)(2), 180.
https://doi.org/10.3390/biom11020180

Lopandic Z, Dragačević L, Popovic D, Andjelkovic U, Minić R, Gavrovic-Jankulovic M. BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms. in Biomolecules. 2021;11(2)(2):180.
doi:10.3390/biom11020180 .

Lopandic, Zorana, Dragačević, Luka, Popovic, Dragan, Andjelkovic, Uros, Minić, Rajna, Gavrovic-Jankulovic, Marija, "BanLec-eGFP Chimera as a Tool for Evaluation of Lectin Binding to High-Mannose Glycans on Microorganisms" in Biomolecules, 11(2), no. 2 (2021):180,
https://doi.org/10.3390/biom11020180 . .

TY  - JOUR
AU  - Dragačević, Luka
AU  - Đorđević, Brižita
AU  - Gavrović-Jankulović, Marija
AU  - Ilić, Vesna
AU  - Kanazir, Danijela
AU  - Minić, Rajna
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/563
AB  - The surface of microorganisms is covered with polysaccharide structures which are in immediate contact with receptor structures on host's cells and antibodies. The interaction between microorganisms and their host is dependent on surface glycosylation and in this study we have tested the interaction of plant lectins with different microorganisms. Enzyme-linked lectin sorbent assay - ELLSA was used to test the binding of recombinant Musa acuminata lectin - BL to 27 selected microorganisms and 7 other lectins were used for comparison: Soy bean agglutinin - SBA, Lens culinaris lectin - LCA, Wheat germ agglutinin - WGA, RCA(120) - Ricinus communis agglutinin, Con A - from Canavalia ensiformis, Sambucus nigra agglutinin - SNA I and Maackia amurensis agglutinin - MAA. The goal was to define the microorganisms' surface glycosylation by means of interaction with the selected plant lectins and to make a comparison with BL. Among the tested lectins most selective binding was observed for RCA(120) which preferentially bound Lactobacillus casei DG. Recombinant banana lectin showed specific binding to all of the tested fungal species. The binding of BL to Candida albicans was further tested with fluorescence microscopy and flow cytometry and it was concluded that this lectin can differentiate ss-glucan rich surfaces. The binding of BL to S. boulardii could be inhibited with ss-glucan from yeast with IC50 1.81 mu g mL(-1) and to P. roqueforti with 1.10 mu g mL(-1). This unique specificity of BL could be exploited for screening purposes and potentially for the detection of ss-glucan in solutions.
PB  - Springer, Dordrecht
T2  - Glycoconjugate Journal
T1  - ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin
EP  - 105
IS  - 1
SP  - 95
VL  - 37
DO  - 10.1007/s10719-019-09898-8
ER  - 

@article{
author = "Dragačević, Luka and Đorđević, Brižita and Gavrović-Jankulović, Marija and Ilić, Vesna and Kanazir, Danijela and Minić, Rajna",
year = "2020",
abstract = "The surface of microorganisms is covered with polysaccharide structures which are in immediate contact with receptor structures on host's cells and antibodies. The interaction between microorganisms and their host is dependent on surface glycosylation and in this study we have tested the interaction of plant lectins with different microorganisms. Enzyme-linked lectin sorbent assay - ELLSA was used to test the binding of recombinant Musa acuminata lectin - BL to 27 selected microorganisms and 7 other lectins were used for comparison: Soy bean agglutinin - SBA, Lens culinaris lectin - LCA, Wheat germ agglutinin - WGA, RCA(120) - Ricinus communis agglutinin, Con A - from Canavalia ensiformis, Sambucus nigra agglutinin - SNA I and Maackia amurensis agglutinin - MAA. The goal was to define the microorganisms' surface glycosylation by means of interaction with the selected plant lectins and to make a comparison with BL. Among the tested lectins most selective binding was observed for RCA(120) which preferentially bound Lactobacillus casei DG. Recombinant banana lectin showed specific binding to all of the tested fungal species. The binding of BL to Candida albicans was further tested with fluorescence microscopy and flow cytometry and it was concluded that this lectin can differentiate ss-glucan rich surfaces. The binding of BL to S. boulardii could be inhibited with ss-glucan from yeast with IC50 1.81 mu g mL(-1) and to P. roqueforti with 1.10 mu g mL(-1). This unique specificity of BL could be exploited for screening purposes and potentially for the detection of ss-glucan in solutions.",
publisher = "Springer, Dordrecht",
journal = "Glycoconjugate Journal",
title = "ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin",
pages = "105-95",
number = "1",
volume = "37",
doi = "10.1007/s10719-019-09898-8"
}

Dragačević, L., Đorđević, B., Gavrović-Jankulović, M., Ilić, V., Kanazir, D.,& Minić, R.. (2020). ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin. in Glycoconjugate Journal
Springer, Dordrecht., 37(1), 95-105.
https://doi.org/10.1007/s10719-019-09898-8

Dragačević L, Đorđević B, Gavrović-Jankulović M, Ilić V, Kanazir D, Minić R. ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin. in Glycoconjugate Journal. 2020;37(1):95-105.
doi:10.1007/s10719-019-09898-8 .

Dragačević, Luka, Đorđević, Brižita, Gavrović-Jankulović, Marija, Ilić, Vesna, Kanazir, Danijela, Minić, Rajna, "ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin" in Glycoconjugate Journal, 37, no. 1 (2020):95-105,
https://doi.org/10.1007/s10719-019-09898-8 . .

TY  - CONF
AU  - Dragačević, Luka
AU  - Đorđević, Brižita
AU  - Gavrović-Jankulović, Marija
AU  - Ilić, Vesna
AU  - Minić, Rajna
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/567
PB  - Lippincott Williams & Wilkins, Philadelphia
C3  - Journal of Clinical Gastroenterology
T1  - Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa
EP  - S13
SP  - S13
VL  - 54
UR  - https://hdl.handle.net/21.15107/rcub_intor_567
ER  - 

@conference{
author = "Dragačević, Luka and Đorđević, Brižita and Gavrović-Jankulović, Marija and Ilić, Vesna and Minić, Rajna",
year = "2020",
publisher = "Lippincott Williams & Wilkins, Philadelphia",
journal = "Journal of Clinical Gastroenterology",
title = "Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa",
pages = "S13-S13",
volume = "54",
url = "https://hdl.handle.net/21.15107/rcub_intor_567"
}

Dragačević, L., Đorđević, B., Gavrović-Jankulović, M., Ilić, V.,& Minić, R.. (2020). Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa. in Journal of Clinical Gastroenterology
Lippincott Williams & Wilkins, Philadelphia., 54, S13-S13.
https://hdl.handle.net/21.15107/rcub_intor_567

Dragačević L, Đorđević B, Gavrović-Jankulović M, Ilić V, Minić R. Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa. in Journal of Clinical Gastroenterology. 2020;54:S13-S13.
https://hdl.handle.net/21.15107/rcub_intor_567 .

Dragačević, Luka, Đorđević, Brižita, Gavrović-Jankulović, Marija, Ilić, Vesna, Minić, Rajna, "Typing of Surface Glycosilation of Microorganisms by Lectins with in House Elisa" in Journal of Clinical Gastroenterology, 54 (2020):S13-S13,
https://hdl.handle.net/21.15107/rcub_intor_567 .

TY  - CONF
AU  - Dragačević, Luka
AU  - Minić, Rajna
AU  - Michalickova, Danica
AU  - Ilić, Vesna
AU  - Đorđević, Brižita
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/568
PB  - Lippincott Williams & Wilkins, Philadelphia
C3  - Journal of Clinical Gastroenterology
T1  - Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults
EP  - S18
SP  - S18
VL  - 54
UR  - https://hdl.handle.net/21.15107/rcub_intor_568
ER  - 

@conference{
author = "Dragačević, Luka and Minić, Rajna and Michalickova, Danica and Ilić, Vesna and Đorđević, Brižita",
year = "2020",
publisher = "Lippincott Williams & Wilkins, Philadelphia",
journal = "Journal of Clinical Gastroenterology",
title = "Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults",
pages = "S18-S18",
volume = "54",
url = "https://hdl.handle.net/21.15107/rcub_intor_568"
}

Dragačević, L., Minić, R., Michalickova, D., Ilić, V.,& Đorđević, B.. (2020). Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults. in Journal of Clinical Gastroenterology
Lippincott Williams & Wilkins, Philadelphia., 54, S18-S18.
https://hdl.handle.net/21.15107/rcub_intor_568

Dragačević L, Minić R, Michalickova D, Ilić V, Đorđević B. Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults. in Journal of Clinical Gastroenterology. 2020;54:S18-S18.
https://hdl.handle.net/21.15107/rcub_intor_568 .

Dragačević, Luka, Minić, Rajna, Michalickova, Danica, Ilić, Vesna, Đorđević, Brižita, "Comparison of Specific IGG and IGA Subclass Levels to Lactobacillus and Streptococcus in Young Healthy Adults" in Journal of Clinical Gastroenterology, 54 (2020):S18-S18,
https://hdl.handle.net/21.15107/rcub_intor_568 .

TY  - JOUR
AU  - Dragačević, Luka
AU  - Đorđević, Brižita
AU  - Gavrović-Jankulović, Marija
AU  - Ilić, Vesna
AU  - Kanazir, Danijela
AU  - Minić, Rajna
PY  - 2020
UR  - http://intor.torlakinstitut.com/handle/123456789/653
AB  - The surface of microorganisms is covered with polysaccharide structures which are in immediate contact with receptor structures on host's cells and antibodies. The interaction between microorganisms and their host is dependent on surface glycosylation and in this study we have tested the interaction of plant lectins with different microorganisms. Enzyme-linked lectin sorbent assay - ELLSA was used to test the binding of recombinant Musa acuminata lectin - BL to 27 selected microorganisms and 7 other lectins were used for comparison: Soy bean agglutinin - SBA, Lens culinaris lectin - LCA, Wheat germ agglutinin - WGA, RCA(120) - Ricinus communis agglutinin, Con A - from Canavalia ensiformis, Sambucus nigra agglutinin - SNA I and Maackia amurensis agglutinin - MAA. The goal was to define the microorganisms' surface glycosylation by means of interaction with the selected plant lectins and to make a comparison with BL. Among the tested lectins most selective binding was observed for RCA(120) which preferentially bound Lactobacillus casei DG. Recombinant banana lectin showed specific binding to all of the tested fungal species. The binding of BL to Candida albicans was further tested with fluorescence microscopy and flow cytometry and it was concluded that this lectin can differentiate ss-glucan rich surfaces. The binding of BL to S. boulardii could be inhibited with ss-glucan from yeast with IC50 1.81 mu g mL(-1) and to P. roqueforti with 1.10 mu g mL(-1). This unique specificity of BL could be exploited for screening purposes and potentially for the detection of ss-glucan in solutions.
PB  - Springer, Dordrecht
T2  - Glycoconjugate Journal
T1  - ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin
EP  - 105
IS  - 1
SP  - 95
VL  - 37
DO  - 10.1007/s10719-019-09898-8
ER  - 

@article{
author = "Dragačević, Luka and Đorđević, Brižita and Gavrović-Jankulović, Marija and Ilić, Vesna and Kanazir, Danijela and Minić, Rajna",
year = "2020",
abstract = "The surface of microorganisms is covered with polysaccharide structures which are in immediate contact with receptor structures on host's cells and antibodies. The interaction between microorganisms and their host is dependent on surface glycosylation and in this study we have tested the interaction of plant lectins with different microorganisms. Enzyme-linked lectin sorbent assay - ELLSA was used to test the binding of recombinant Musa acuminata lectin - BL to 27 selected microorganisms and 7 other lectins were used for comparison: Soy bean agglutinin - SBA, Lens culinaris lectin - LCA, Wheat germ agglutinin - WGA, RCA(120) - Ricinus communis agglutinin, Con A - from Canavalia ensiformis, Sambucus nigra agglutinin - SNA I and Maackia amurensis agglutinin - MAA. The goal was to define the microorganisms' surface glycosylation by means of interaction with the selected plant lectins and to make a comparison with BL. Among the tested lectins most selective binding was observed for RCA(120) which preferentially bound Lactobacillus casei DG. Recombinant banana lectin showed specific binding to all of the tested fungal species. The binding of BL to Candida albicans was further tested with fluorescence microscopy and flow cytometry and it was concluded that this lectin can differentiate ss-glucan rich surfaces. The binding of BL to S. boulardii could be inhibited with ss-glucan from yeast with IC50 1.81 mu g mL(-1) and to P. roqueforti with 1.10 mu g mL(-1). This unique specificity of BL could be exploited for screening purposes and potentially for the detection of ss-glucan in solutions.",
publisher = "Springer, Dordrecht",
journal = "Glycoconjugate Journal",
title = "ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin",
pages = "105-95",
number = "1",
volume = "37",
doi = "10.1007/s10719-019-09898-8"
}

Dragačević, L., Đorđević, B., Gavrović-Jankulović, M., Ilić, V., Kanazir, D.,& Minić, R.. (2020). ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin. in Glycoconjugate Journal
Springer, Dordrecht., 37(1), 95-105.
https://doi.org/10.1007/s10719-019-09898-8

Dragačević L, Đorđević B, Gavrović-Jankulović M, Ilić V, Kanazir D, Minić R. ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin. in Glycoconjugate Journal. 2020;37(1):95-105.
doi:10.1007/s10719-019-09898-8 .

Dragačević, Luka, Đorđević, Brižita, Gavrović-Jankulović, Marija, Ilić, Vesna, Kanazir, Danijela, Minić, Rajna, "ELLSA based profiling of surface glycosylation in microorganisms reveals that ss-glucan rich yeasts' surfaces are selectively recognized with recombinant banana lectin" in Glycoconjugate Journal, 37, no. 1 (2020):95-105,
https://doi.org/10.1007/s10719-019-09898-8 . .

TY  - JOUR
AU  - Pavlović, Bojana
AU  - Cvijetić, Nataša
AU  - Dragačević, Luka
AU  - Ivković, Branka
AU  - Vujić, Zorica
AU  - Kuntić, Vesna
PY  - 2016
UR  - http://intor.torlakinstitut.com/handle/123456789/469
AB  - One of the most commonly used surfactants in the production of split virus influenza vaccine is nonionic surfactant Triton X-100. After splitting of the virus is accomplished, Triton X-100 is removed from the vaccine by subsequent production steps. Because of toxicity of Triton X-100, which remains in the vaccine in residual amounts, a sufficiently sensitive method for its detection and quantification needs to be defined. Two methods for determination of Triton X-100 residuals were developed: the UV-spectrophotometry and HPLC methods. For both methods, preparation of vaccine samples and removal of proteins and virus particles were crucial: samples were treated with methanol (1:1) and then centrifuged at 25000 x g for 30 min. After such treatment, the majority of vaccine components that interfered in the UV region were removed, and diluted samples could be directly measured. The chromatographic system included C18 column, step methanol gradient, and detection at 225 nm with a single peak of Triton X-100 at 12.6 min. Both methods were validated and gave satisfactory results for accuracy, precision, specificity, linearity, and robustness. LOQ was slightly lower for the HPLC method. Hence, it was shown that both methods are suitable for analysis of residual amounts of Triton X-100, with the advantages of the UV method being its simplicity and availability in most laboratories.
PB  - Oxford Univ Press Inc, Cary
T2  - Journal of AOAC International
T1  - Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine
EP  - 400
IS  - 2
SP  - 396
VL  - 99
DO  - 10.5740/jaoacint.15-0201
ER  - 

@article{
author = "Pavlović, Bojana and Cvijetić, Nataša and Dragačević, Luka and Ivković, Branka and Vujić, Zorica and Kuntić, Vesna",
year = "2016",
abstract = "One of the most commonly used surfactants in the production of split virus influenza vaccine is nonionic surfactant Triton X-100. After splitting of the virus is accomplished, Triton X-100 is removed from the vaccine by subsequent production steps. Because of toxicity of Triton X-100, which remains in the vaccine in residual amounts, a sufficiently sensitive method for its detection and quantification needs to be defined. Two methods for determination of Triton X-100 residuals were developed: the UV-spectrophotometry and HPLC methods. For both methods, preparation of vaccine samples and removal of proteins and virus particles were crucial: samples were treated with methanol (1:1) and then centrifuged at 25000 x g for 30 min. After such treatment, the majority of vaccine components that interfered in the UV region were removed, and diluted samples could be directly measured. The chromatographic system included C18 column, step methanol gradient, and detection at 225 nm with a single peak of Triton X-100 at 12.6 min. Both methods were validated and gave satisfactory results for accuracy, precision, specificity, linearity, and robustness. LOQ was slightly lower for the HPLC method. Hence, it was shown that both methods are suitable for analysis of residual amounts of Triton X-100, with the advantages of the UV method being its simplicity and availability in most laboratories.",
publisher = "Oxford Univ Press Inc, Cary",
journal = "Journal of AOAC International",
title = "Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine",
pages = "400-396",
number = "2",
volume = "99",
doi = "10.5740/jaoacint.15-0201"
}

Pavlović, B., Cvijetić, N., Dragačević, L., Ivković, B., Vujić, Z.,& Kuntić, V.. (2016). Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine. in Journal of AOAC International
Oxford Univ Press Inc, Cary., 99(2), 396-400.
https://doi.org/10.5740/jaoacint.15-0201

Pavlović B, Cvijetić N, Dragačević L, Ivković B, Vujić Z, Kuntić V. Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine. in Journal of AOAC International. 2016;99(2):396-400.
doi:10.5740/jaoacint.15-0201 .

Pavlović, Bojana, Cvijetić, Nataša, Dragačević, Luka, Ivković, Branka, Vujić, Zorica, Kuntić, Vesna, "Direct UV Spectrophotometry and HPLC Determination of Triton X-100 in Split Virus Influenza Vaccine" in Journal of AOAC International, 99, no. 2 (2016):396-400,
https://doi.org/10.5740/jaoacint.15-0201 . .