Abstract

LITWIN, Silvana; ISABETTINI, Alberto Oscar; CALDERON, Leandro  and  VARNI, Liliana Mónica. Optimization in the immunoglobulins recovery, as whole molecule and F(ab´)₂ fragment, in the antivenim production . Acta toxicol. argent. [online]. 2014, vol.22, n.2, pp.82-89. ISSN 1851-3743.

Today two methods are mainly used for the purification of antibodies from hyperimmune equine plasma at industrial level obtaining enriched preparations of immunoglobulin G (IgG) molecules or F(ab´)₂ fragments. In both methods, after the precipitation, an important loss in the neutralizing capability was observed compared to the one of the original plasma. In this work, we performed the fractionation of hyperimmune equine plasma using caprylic acid, with and without enzymatic digestion with pepsin. The aim was to explain the percentage of recovery of the neutralizing capability after the fractionation; which resulted minor when the plasma was enzymatically treated. Additionally, we intended to establish which stage, in the purification process, was the responsible for the difference in the antibody recovery between one methodology and the other. When entire immunoglobulins were purified, approximately 53% of the neutralizing capacity was recovered, but when the sample was purified after the enzymatic treatment, around the 30% of the activity was obtained. A ratio of similar magnitude is verified on the recovery of the soluble protein mass after the removal of contaminants, between the two methods. The insolubilization of the fragment Fc generated during digestion would be responsible for the additional loss of protein and neutralizing capacity.

Keywords : Antivenom; Recovery; Caprylic acid; Immunoglobulin.