Purification of F(ab′)2 anti-snake venom by caprylic acid: A fast method for obtaining IgG fragments with high neutralization activity, purity and yield

M. C. dos Santos, M. R. D'Império Lima, G. C. Furtado, G. M.D.D. Colletto, T. L. Kipnis, W. Dias da Silva

Research output: Contribution to journalArticlepeer-review

68 Scopus citations

Abstract

Pooled horse plasma containing antibodies against Crotalus durissus terrificus whole venom were digested with pepsin at an enzyme-substrate ratio of 8:1, pH 3.1, for 40 min and the F(ab′)2M fragments purified by adding 8.7% caprylic acid (pH 5.0). For comparison, F(ab′)2B purified by precipitation with ammonium sulphate and uncleaved IgG purified with caprylic acid were also prepared. Fab′ fragments were obtained by reduction and alkylation of F(ab′)2B. The anti-whole C. d. terrificus venom titers, determined by Dot-Blot were 12,800 (IgG), 6400 [F(ab′)2B], 4800 [F(ab′)2M] and 3200 (Fab′B). Immunochemical analysis of these fragments by SDS gel electrophoresis, Western blot and by double immunodiffusion revealed that the solution containing. F(ab′)2M was free of IgG and of other plasma proteins, whereas that containing F(ab′)2B was not. One milligram of either F(ab′)2B, F(ab′)2M or Fab′B was able to neutralize respectively 20.7 μg, 20.2 μg and 13.8 μg of C. d. terrificus venom.

Original languageEnglish
Pages (from-to)297-303
Number of pages7
JournalToxicon
Volume27
Issue number3
DOIs
StatePublished - 1989
Externally publishedYes

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