Separation, purification and identification of human apolipoprotein H

Objective To explore a method to separate and purify conveniently and efficiently apolipoprotein H from human plasma. Methods We added perchloric acid into human plasma. Apolipoprotein H was separated and purified by ion exchange chromatography and affinity chromatography. The purified apolipoprotein H was analyzed by SDS-polyacrylamide gel electrophoresis, Western blot and BCA kit. Results We repeated the experiment three times. The purity of the apolipoprotein H was all over 98%. Western blot demonstrated that anti β₂- Glycoprotein I monoclonal antibody reacted with the purified apolipoprotein H. We obtained a total of 12. 56 mg purified apolipoprotein H, average 21 mg/L. Conclusions This method has good repeatability, reliability and more convenient and economical than the traditional method.