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The antibody fragments generated from hyperimmune equine IgG is widely used as anti-snake venom, anti-scorpion venom, anti-diphtheria, anti-tetanus, anti-gangrene and anti-rabies agents. Antibody fragments, F(ab) 2 , because of their specificity and absence of undesired reactivity are preferred over complete IgG. This paper discusses a novel purification technique for chromatographic purification of anti-rabies immunoglobulin G (IgG) fragment F(ab) 2 from horse serum. F(ab) 2 was purified by two successive chromatography steps using Cellufine A-200 and ProSep-vA Ultra media. The purified F(ab) 2 was characterized using biochemical and biophysical methods and shown to be pure and homogeneous. The purified F(ab) 2 was reactive to rabies antigen in immuno-electrophoresis and diffusion tests. The purified F(ab) 2 was biologically functional and was found to show a potency of 1500 IU ml −1 . Comparative analysis of the purity with commercially available F(ab) 2 by HPLC analysis and SDS–PAGE indicated that the present product is better in purity. To our knowledge, this is the first report providing evidence on purification of equine antibody fragment using controlled pore glass based protein A chromatography media.

The method for the determination of caprylic acid and sodium caprylate from biological products was systematically validated using NEFA-C kit. The results obtained demonstrated that the kit method was simple, rapid, reliable, sensitive, reproducible and cost effective in comparison to the current methods i.e. colorimetric, High Performance Liquid Chromatography (HPLC) and Gas Chromatography (GC) methods. The assay exhibited excellent linearity, accuracy, precision and robustness. Mean recoveries ranged between 95 and 101.3% (n=6). The proposed method was linear over the concentration range of 0.05-10mM of caprylate with values of coefficient of regression being>0.99. Method showed sensitivity of 0.05 mM (7.21 microg/ml for caprylic acid and 8.31 microg/ml for sodium caprylate). The % Relative standard Deviation (%RSD) for intra and interprecision studies was less than 5%. In conclusion the validated method was successfully used in monitoring of processed bulk and final products generated during production of biological products thus laying emphasis on strict control of release criteria for biological products fractionated using caprylic acid.