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
Ćupić, Vitomir
Borozan, Sunčica
Nešić, Andrijana
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
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
Колекције
Институција/група
Fakultet veterinarske medicineTY - JOUR AU - Milovanović, Vladimir AU - Minić, Rajna AU - Vakić, Jelena AU - Ivanović, Saša AU - Ćupić, Vitomir AU - Borozan, Sunčica AU - Nešić, Andrijana AU - Živković, Irena PY - 2021 UR - https://vet-erinar.vet.bg.ac.rs/handle/123456789/1994 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 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 Ćupić, Vitomir and Borozan, Sunčica and Nešić, Andrijana 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", 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., Ćupić, 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., 192, 57-65. https://doi.org/10.1016/j.toxicon.2021.01.012
Milovanović V, Minić R, Vakić J, Ivanović S, Ćupić 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, Jelena, Ivanović, Saša, Ćupić, Vitomir, Borozan, Sunčica, Nešić, Andrijana, 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 . .