TY  - JOUR
AU  - Lukić, Ivana
AU  - Dimitrijević, Ljiljana
AU  - Dovezenski, Nebojsa
AU  - Živković, Irena
AU  - Petrušić, Vladimir
AU  - Marinković, Emilija
AU  - Inić-Kanada, Aleksandra
AU  - Stojanović, Marijana
PY  - 2011
UR  - http://intor.torlakinstitut.com/handle/123456789/319
AB  - Given an existing demand to establish a process of tetanus vaccine production in a way that allows its complete validation and standardization, this paper focuses on tetanus toxoid purification step. More precisely, we were looking at a possibility to replace the widely used ammonium-sulphate precipitation by a chromatographic method. Based on the tetanus toxin's biochemical characteristics, we have decided to examine the possibility of tetanus toxoid purification by hydrophobic chromatography, and by chromatographic techniques based on interaction with immobilized metal ions, i.e. chelating chromatography and immobilized metal affinity chromatography. We used samples obtained from differently fragmented crude tetanus toxins by formaldehyde treatment (assigned as TTd-A and TTd-B) as starting material for tetanus toxoid purification. Obtained results imply that purification of tetanus toxoid by hydrophobic chromatography represents a good alternative to ammonium-sulphate precipitation. Tetanus toxoid preparations obtained by hydrophobic chromatography were similar to those obtained by ammonium-sulphate precipitation in respect to yield, purity and immunogenicity. In addition, their immunogenicity was similar to standard tetanus toxoid preparation (NIBSC, Potters Bar, UK). Furthermore, the characteristics of crude tetanus toxin preparations had the lowest impact on the final purification product when hydrophobic chromatography was the applied method of tetanus toxoid purification. On the other hand, purifications of tetanus toxoid by chelating chromatography or immobilized metal affinity chromatography generally resulted in a very low yield due to not satisfactory tetanus toxoid binding to the column, and immunogenicity of the obtained tetanus toxoid-containing preparations was poor. (C) 2011 Elsevier BM. All rights reserved.
PB  - Elsevier Science Bv, Amsterdam
T2  - Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
T1  - Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation
EP  - 2219
IS  - 23
SP  - 2213
VL  - 879
DO  - 10.1016/j.jchromb.2011.06.003
ER  - 

@article{
author = "Lukić, Ivana and Dimitrijević, Ljiljana and Dovezenski, Nebojsa and Živković, Irena and Petrušić, Vladimir and Marinković, Emilija and Inić-Kanada, Aleksandra and Stojanović, Marijana",
year = "2011",
abstract = "Given an existing demand to establish a process of tetanus vaccine production in a way that allows its complete validation and standardization, this paper focuses on tetanus toxoid purification step. More precisely, we were looking at a possibility to replace the widely used ammonium-sulphate precipitation by a chromatographic method. Based on the tetanus toxin's biochemical characteristics, we have decided to examine the possibility of tetanus toxoid purification by hydrophobic chromatography, and by chromatographic techniques based on interaction with immobilized metal ions, i.e. chelating chromatography and immobilized metal affinity chromatography. We used samples obtained from differently fragmented crude tetanus toxins by formaldehyde treatment (assigned as TTd-A and TTd-B) as starting material for tetanus toxoid purification. Obtained results imply that purification of tetanus toxoid by hydrophobic chromatography represents a good alternative to ammonium-sulphate precipitation. Tetanus toxoid preparations obtained by hydrophobic chromatography were similar to those obtained by ammonium-sulphate precipitation in respect to yield, purity and immunogenicity. In addition, their immunogenicity was similar to standard tetanus toxoid preparation (NIBSC, Potters Bar, UK). Furthermore, the characteristics of crude tetanus toxin preparations had the lowest impact on the final purification product when hydrophobic chromatography was the applied method of tetanus toxoid purification. On the other hand, purifications of tetanus toxoid by chelating chromatography or immobilized metal affinity chromatography generally resulted in a very low yield due to not satisfactory tetanus toxoid binding to the column, and immunogenicity of the obtained tetanus toxoid-containing preparations was poor. (C) 2011 Elsevier BM. All rights reserved.",
publisher = "Elsevier Science Bv, Amsterdam",
journal = "Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences",
title = "Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation",
pages = "2219-2213",
number = "23",
volume = "879",
doi = "10.1016/j.jchromb.2011.06.003"
}

Lukić, I., Dimitrijević, L., Dovezenski, N., Živković, I., Petrušić, V., Marinković, E., Inić-Kanada, A.,& Stojanović, M.. (2011). Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation. in Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
Elsevier Science Bv, Amsterdam., 879(23), 2213-2219.
https://doi.org/10.1016/j.jchromb.2011.06.003

Lukić I, Dimitrijević L, Dovezenski N, Živković I, Petrušić V, Marinković E, Inić-Kanada A, Stojanović M. Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation. in Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences. 2011;879(23):2213-2219.
doi:10.1016/j.jchromb.2011.06.003 .

Lukić, Ivana, Dimitrijević, Ljiljana, Dovezenski, Nebojsa, Živković, Irena, Petrušić, Vladimir, Marinković, Emilija, Inić-Kanada, Aleksandra, Stojanović, Marijana, "Tetanus toxoid purification: Chromatographic procedures as an alternative to ammonium-sulphate precipitation" in Journal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences, 879, no. 23 (2011):2213-2219,
https://doi.org/10.1016/j.jchromb.2011.06.003 . .