Development of an ELISA to assess the potency of horse therapeutic polyvalent antibothropic antivenom.
The objective of this study was the search for a suitable venom antigen to be used in an in vitro alternative immunoassay, to the standard antivenom neutralization assay using mice. Bothrops jararaca venom was fractionated in DEAE-Sephacel columns and the fractions were tested for a correlation between antibody capture enzyme linked immunosorbent assay (ELISA) absorbance values and the 'in vivo' antivenom potency. Individual antivenoms from 14 horses and 15 separate FUNED polyspecific Bothrops ampouled antivenoms (final product) were used. Fractions showing the higher correlations were further chromatographed in a Sephadex G-75 column and again tested for the correlation. Two fractions with haemorrhagic activity displayed a correlation of r = 0.77 and r = 0.8 against the individual horse antivenom sera and of r = 0.79 and r = 0.8 for the ampouled antivenom. For all results p < 0.001. Two other fractions with phospholipase A2 activity showed a correlation of r = 0.66 (p < 0.01) and r = 0.56 (p < 0.03) against the individual horse antivenom sera. Electrophoresis results show a similar composition for both antigens with haemorrhagic activity. Results indicate that the fractions purified would be suitable for the desired objective of this study.
Heneine
L G
LG
Centro de Pesquisa e Desenvolvimento, Fundação Ezequiel Dias, Belo Horizonte, Brazil.
Carvalho
A D
AD
Barbosa
C F
CF
Arávjo dos Santos
M R
MR
D000818:Animals; D000997:Antivenins / Q000276:immunology*; D001777:Blood Coagulation / Q000187:drug effects; D017837:Bothrops; D005591:Chemical Fractionation; D002852:Chromatography, Ion Exchange; D003435:Crotalid Venoms / Q000737:chemistry / Q000276:immunology* / Q000633:toxicity; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D006736:Horses; D007928:Lethal Dose 50; D051379:Mice; D009500:Neutralization Tests; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2
D000997:Antivenins; D003435:Crotalid Venoms; D010741:Phospholipases A; D054467:Phospholipases A2