MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation
Само за регистроване кориснике
2021
Аутори
Milovanović, VladimirMinić, Rajna
Vakić, J.
Ivanović, S.
Cupić, V.
Borozan, S.
Nešić, A.
Živković, Irena
Чланак у часопису (Објављена верзија)
Метаподаци
Приказ свих података о документуАпстракт
The MTT assay is routinely used to detect the activity of living cells. While working with Vipera ammodytes venom we detected the reduction of MTT without the presence of cells, in a concentration-dependent manner. By combining non-reducing PAGE, L-amino acid oxidase (LAAO) assays, and standard MTT assays, we established and confirmed that venom MTT reduction is catalyzed by only one enzyme, the LAAO. Even though it was previously known that the dimeric and tetrameric forms of LAAO are active, we conclude that the enzyme is also active in the monomeric form. Our results have led to the definition of a new MTT assay in a microtiter plate for in vitro testing of svLAAO activity i.e. from the venom of the V. ammodytes snake. Potentially, this method can be used for testing hemorrhagic venoms of other snakes as well as the LAAO neutralization capability of appropriate antivenoms.
Кључне речи:
Antivenom / L-Amino acid Oxidase monomer / MTT / Potency test / Vipera ammodytes venomИзвор:
Toxicon, 2021, 192, 57-65Издавач:
- Elsevier Ltd
Финансирање / пројекти:
DOI: 10.1016/j.toxicon.2021.01.012
ISSN: 0041-0101
WoS: 000623933000006
Scopus: 2-s2.0-85099912728
Институција/група
TorlakTY - JOUR AU - Milovanović, Vladimir AU - Minić, Rajna AU - Vakić, J. AU - Ivanović, S. AU - Cupić, V. AU - Borozan, S. AU - Nešić, A. AU - Živković, Irena PY - 2021 UR - http://intor.torlakinstitut.com/handle/123456789/572 AB - The MTT assay is routinely used to detect the activity of living cells. While working with Vipera ammodytes venom we detected the reduction of MTT without the presence of cells, in a concentration-dependent manner. By combining non-reducing PAGE, L-amino acid oxidase (LAAO) assays, and standard MTT assays, we established and confirmed that venom MTT reduction is catalyzed by only one enzyme, the LAAO. Even though it was previously known that the dimeric and tetrameric forms of LAAO are active, we conclude that the enzyme is also active in the monomeric form. Our results have led to the definition of a new MTT assay in a microtiter plate for in vitro testing of svLAAO activity i.e. from the venom of the V. ammodytes snake. Potentially, this method can be used for testing hemorrhagic venoms of other snakes as well as the LAAO neutralization capability of appropriate antivenoms. PB - Elsevier Ltd T2 - Toxicon T1 - MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation EP - 65 SP - 57 VL - 192 DO - 10.1016/j.toxicon.2021.01.012 ER -
@article{ author = "Milovanović, Vladimir and Minić, Rajna and Vakić, J. and Ivanović, S. and Cupić, V. and Borozan, S. and Nešić, A. and Živković, Irena", year = "2021", abstract = "The MTT assay is routinely used to detect the activity of living cells. While working with Vipera ammodytes venom we detected the reduction of MTT without the presence of cells, in a concentration-dependent manner. By combining non-reducing PAGE, L-amino acid oxidase (LAAO) assays, and standard MTT assays, we established and confirmed that venom MTT reduction is catalyzed by only one enzyme, the LAAO. Even though it was previously known that the dimeric and tetrameric forms of LAAO are active, we conclude that the enzyme is also active in the monomeric form. Our results have led to the definition of a new MTT assay in a microtiter plate for in vitro testing of svLAAO activity i.e. from the venom of the V. ammodytes snake. Potentially, this method can be used for testing hemorrhagic venoms of other snakes as well as the LAAO neutralization capability of appropriate antivenoms.", publisher = "Elsevier Ltd", journal = "Toxicon", title = "MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation", pages = "65-57", volume = "192", doi = "10.1016/j.toxicon.2021.01.012" }
Milovanović, V., Minić, R., Vakić, J., Ivanović, S., Cupić, V., Borozan, S., Nešić, A.,& Živković, I.. (2021). MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation. in Toxicon Elsevier Ltd., 192, 57-65. https://doi.org/10.1016/j.toxicon.2021.01.012
Milovanović V, Minić R, Vakić J, Ivanović S, Cupić V, Borozan S, Nešić A, Živković I. MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation. in Toxicon. 2021;192:57-65. doi:10.1016/j.toxicon.2021.01.012 .
Milovanović, Vladimir, Minić, Rajna, Vakić, J., Ivanović, S., Cupić, V., Borozan, S., Nešić, A., Živković, Irena, "MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation" in Toxicon, 192 (2021):57-65, https://doi.org/10.1016/j.toxicon.2021.01.012 . .