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Using the caprylic acid in obtaining the horse immunoglobulin anti tetanus toxin

Abstract:

Purified horse anti tetanus toxin immunoglobulins with high antibody concentration and biological activity, are used as heterologous sera and standard serum for corresponding vaccine. However, the selection of precipitating agents for the purification of them is a problem in terms of purity, yield and economy, which still remains to be solved. In this study, a procedure to fractionation horse plasma anti tetanus toxin by Caprylic acid precipitation is described. A solution of acetic acid/sodium acetate was added to the samples to achieve different ionic strengths, 0.05-0.4 moles/L, and caprylic acid was added to each volume to reach final concentrations from 1 to 7% (v / v), with vigorous stirring for 60 min. Non-immunoglobulin proteins precipitated under these conditions, while immunoglobulins remained in the supernatant, which was then diafiltered to remove caprylic acid and concentrate inmunoglobulins. This methodology was compared to that based on ammonium sulfate fractionation. The best results were given when caprylic acid was added to plasma, until a final caprylic acid concentration of 3% was reached, at ionic strength 0.2 moles/L and pH adjusted to 4.5. The IgG recovery was 91-95% by using the caprylic acid method. It was achieved a protein concentration of 27.1-29.3 mg/mL and an albumin/immunoglobulins ratio of 0.019-0.021. The purity was 91- 95% and the anti-tetanus toxin activity recovery of 93-96%.