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.
TY - 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",
url = "http://intor.torlakinstitut.com/handle/123456789/572",
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.
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. Toxicon. 2021;192:57-65
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" Toxicon, 192 (2021):57-65,
https://doi.org/10.1016/j.toxicon.2021.01.012 .
