Immunochemistry of human plasma high density lipoproteins. Radioimmunoassay of Apolipoprotein A-II

Apolipoprotein A-II (ApoA-II) is a major apoprotein of human plasma high density lipoproteins (HDL). The apoprotein has two identical chains of known amino acid sequence; the chains are linked by a single disulfide bond. In the present study, we have developed a specific radioimmunoassay for apoA-II that provides a convenient and reproducible method for measuring 10-100 ng of apoprotein. The assay was based on the competition of apoprotein with isotopically labeled [125I]apoA-II. Dioxane (52 percent) was used to separate antibody bound [125I]apoA-II from the free apoprotein. The assay has enabled us to begin studies directed toward mapping-out of the antigenic reactive regions of apoA-II. It has also allowed us to determine the interrelationship between the lipid-binding sites and the antigenic sites of the molecule. The antigenic reactivity of apoA-II was approximately the same in HDL and phospholipid-apoA-II complexes as that of the free apoprotein. However, succinylation of apoA-II was associated with a marked decrease in antigenic reactivity