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MTT based L-aminoacid oxidase activity test for determination of antivenom potency against Vipera ammodytes envenomation

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2021
Authors
Milovanović, Vladimir
Minić, Rajna
Vakić, J.
Ivanović, S.
Cupić, V.
Borozan, S.
Nešić, A.
Živković, Irena
article (publishedVersion)
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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.
Keywords:
Antivenom / L-Amino acid Oxidase monomer / MTT / Potency test / Vipera ammodytes venom
Source:
Toxicon, 2021, 192, 57-65
Publisher:
  • Elsevier Ltd
Funding / projects:
  • info:eu-repo/grantAgreement/MESTD/inst-2020/200177/RS// (RS-200177)

DOI: 10.1016/j.toxicon.2021.01.012

ISSN: 0041-0101

WoS: 000623933000006

Scopus: 2-s2.0-85099912728
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URI
http://intor.torlakinstitut.com/handle/123456789/572
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