Polyvalent horse F(Ab`)2 snake antivenom: Development of process to produce polyvalent horse F(Ab`)2 antibodies anti-african snake venom

摘要

A method to obtain polyvalent anti-Bitis?and polyvalent-anti-Naja?antibodies was developed by immunizing horses with?B. arietans,?B. nasicornis,?B. rhinoceros,?N. melanoleuca?and?N. mossambica?crude venoms. Antibody production was followed by the ELISA method during the immunization procedure. Once the desired anti-venom antibody titers were attained, horses were bled and the immunoglobulins were separated from the sera by (NH4)2SO4?precipitation, cleaved with pepsin and filtered through a 30 kDa ultrafiltration membrane. F(ab′)2?fragments were further purified by Q-Fast Flow chromatography, concentrated by molecular ultrafiltration and sterilized by filtration through 0.22 μm membranes. The resulting F(ab′)2preparations were rich in intact L and in pieces of H IgG(T) chains, as demonstrated by electrophoresis and Western blot and exhibited high antibody titers, as assayed by the ELISA method. In addition, the preparations possess a significant capacity to neutralize the lethality of venoms, as estimated by ED50?determination in mouse assay and are free of toxic substances, pyrogen and bacterial or fungal contaminations.