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 antige n to be used in an in vitro alternative immunoassay, to the standard a ntivenom neutralization assay using mice. Bothrops jararaca venom was fractionated in DEAE-Sephacel columns and the fractions were tested fo r a correlation between antibody capture enzyme linked immunosorbent a ssay (ELISA) absorbance values and the 'in vivo' antivenom potency, In dividual antivenoms from 14 horses and 15 separate FUNED polyspecific Bothrops ampouled antivenoms (final product) were used. Fractions show ing 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 fra ctions with phospholipase A(2) activity showed a correlation of r = 0. 66 (p < 0.01) and r = 0.56 (p < 0.03) against the individual horse ant ivenom sera. Electrophoresis results show a similar composition for bo th antigens with haemorrhagic activity. Results indicate that the frac tions purified would be suitable for the desired objective of this stu dy. (C) 1998 Elsevier Science Ltd. All rights reserved.