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, Jelena
Ivanović, Saša
Čupić, Vitomir
Borozan, Sunćica
Nešić, Andrijana N.
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 / venomИзвор:
Toxicon, 2021, 192, 57-65Издавач:
- Elsevier
Финансирање / пројекти:
- Министарство науке, технолошког развоја и иновација Републике Србије, институционално финансирање - 200177 (Центар за имунолошка истраживања 'Бранислав Јанковић' Торлак, Београд) (RS-MESTD-inst-2020-200177)
DOI: 10.1016/j.toxicon.2021.01.012
ISSN: 0041-0101
WoS: 000623933000006
Scopus: 2-s2.0-85099912728
URI
https://www.sciencedirect.com/science/article/pii/S0041010121000362https://cherry.chem.bg.ac.rs/handle/123456789/4490
Колекције
Институција/група
Hemijski fakultet / Faculty of ChemistryTY - JOUR AU - Milovanović, Vladimir AU - Minić, Rajna AU - Vakić, Jelena AU - Ivanović, Saša AU - Čupić, Vitomir AU - Borozan, Sunćica AU - Nešić, Andrijana N. AU - Živković, Irena PY - 2021 UR - https://www.sciencedirect.com/science/article/pii/S0041010121000362 UR - https://cherry.chem.bg.ac.rs/handle/123456789/4490 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 T2 - Toxicon T2 - ToxiconToxicon T1 - MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation VL - 192 SP - 57 EP - 65 DO - 10.1016/j.toxicon.2021.01.012 ER -
@article{ author = "Milovanović, Vladimir and Minić, Rajna and Vakić, Jelena and Ivanović, Saša and Čupić, Vitomir and Borozan, Sunćica and Nešić, Andrijana N. 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", journal = "Toxicon, ToxiconToxicon", title = "MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation", volume = "192", pages = "57-65", doi = "10.1016/j.toxicon.2021.01.012" }
Milovanović, V., Minić, R., Vakić, J., Ivanović, S., Čupić, V., Borozan, S., Nešić, A. N.,& Živković, I.. (2021). MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation. in Toxicon Elsevier., 192, 57-65. https://doi.org/10.1016/j.toxicon.2021.01.012
Milovanović V, Minić R, Vakić J, Ivanović S, Čupić V, Borozan S, Nešić AN, 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, Jelena, Ivanović, Saša, Čupić, Vitomir, Borozan, Sunćica, Nešić, Andrijana N., 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 . .