Isolation and identification of advanced oxidation protein products / 第三军医大学学报

ABSTRACT

Objective To isolate and identify advanced oxidation protein products from human serum albumin (AOPP-HSA), expecting to search for a method of preparing highly purified and bioactive AOPP-HSA. Methods AOPP-HSA crude products were prepared in vitro by exposing HSA to HOCl. AOPP-HSA was isolated by gel chromatography and ion-exchange HPLC. Its structural features and biological activities were characterized by UV and fluorescence spectrum, SDS-PAGE, and the experiment of TNF-? release from monocytes. Results The isolated protein was purified up to 99.4% and was dityrosine-containing protein cross-linking products with molecular weight of 700?10 3. It possessed the ability of triggering the considerable release of TNF-? from monocytes. Conclusion Highly purified and bioactive AOPP-HSA can be successfully prepared by above-mentioned two-step chromatography from AOPP-HSA crude products, which builds a basis for further study of AOPP.