Optimization of Antibody Isolation Collected From Hyperimunized Rabbit using Ammonium SulphateWith and Without Centrifugation

Abstract

Antibody utilization for many purposes are increased nowadays especially in medical applications such as desease therapy and biochemical asssay for diagnosis as well as in research applications. Antibody produced in animal previously purified by various process in order to achieve good quality including isolation, purification and poolishing step. This study aim at gaining optimal condition for antibody isolation from rabbit serum after hyperimunization. Hyperimun serum collected from rabbit immunized by HCG was isolated using various concentration of ammonium sulfat from 20% to 80% with centrifugation while another serum collected from rabbit immunized by IgY was isolated without centrifugation. Pellets of each groups were dialysed and characterized by SDS-PAGE. We found that optimum concentration of ammonium sulphate to isolate serum antibody with centrifugation was achieved at 60%. Additionally, we found that optimum concentration to isolate antibody without centrifugation was achieved at 40% after incubation for 8 hours stepwise.