APPLICATION OF ION EXCHANGE CHROMATOGRAPHY TECHNIQUE TO THE PURIFIED PROCESS TO ENHANCE THE PURITY OF ANTI-RABIES SERUM

Abstract

Anti-rabies serum is indicated for cases of bites from suspected rabid animals at level III or cases of exposure to the rabies virus. This study aims to evaluate the purity of purified anti-rabies serum (SAR) products from the Institute of Vaccines and Medical Biologicals (IVAC) after purification by anion exchange chromatography using Fractogel EMD-DEAE gel. Six lots of anti-rabies serum were purified according to the current process until the dialysis stage and then chromatographed on Fractogel EMD-DEAE gel. Products before and after chromatography were evaluated for purity by SDS-PAGE and SE-HPLC. The purity of SAR before and after chromatography and the performance of the process were evaluated based on protein content, neutralizing antibody, and endotoxin content. The products after chromatography only has the presence of proteins with a molecular weight of 100-135 kDa, equivalent to the MW of the F(ab')₂ antibody fragments, and the ratio of target proteins (F(ab')₂) accounts for 97%-99%. The chromatography process removed over 45% of impurity proteins and achieved an efficiency of over 78%. Using Fractogel EDM-DEAE gel in the SAR serum purification anion exchange chromatography process has created a serum product with higher purity than currently available.