Abstract
Apoliooprotein A-II (apoA-II) was isolated from delipidated human serum high densit lipoprotein (HDL).HDL3 (density1.12-1.21) was preplred from fasting plasma of normolipidemic donors by sequential ultracentrifugation and fractionated on Sepharose 6B column chromatography.HDL3 wasdelipidated with ether-methanol (2: 1v/v) at tempera ture below -20°.Apolipoprotein A-II was then purified by DEAE-Sepharose CL 6B ion exchange column chromatography and Sephadex G-75 chromatogrlphy in the presence of 6M urea.The purity of the protein was ascertained by analytical polyacryamide gel electrophoresis in the presence of 6M urea or 0.1%SDS, amino acid analysis and immunochemical method.Purified apoA-II had molecular weight of 17,000 daltons by gel filtration on a Sephldex G-100 column and 18,000 by SDS polyacrylamide gel electrphorphoresis (without any reducing agent).The amino acid composition of purified apoA-II was consistent with the results previously reported.Antiserum against purified apoA-II was produced in rabbits and immunological properties of the antiserum were characterized.
