Immunochemistry of human plasma apolipoprotein C-II as studied by radioimmunoassay.

ABSTRACT

The immunoreactivity of human plasma apolipoprotein C-II was investigated using a specific radioimmunoassay. In whole plasma, the mean value quantitated was 2.21 +/- 0.415 mg/dl, while in delipidated plasma, a mean value of 3.84 +/- 1.186 mg/dl was obtained, suggesting that the antigenic sites of the apolipoprotein were not fully detected in unmodified plasma by our antibody preparation. Two detergents, Tween-20 and Triton X-100, were studied to determine if they could enhance the immunoreactivity of apolipoprotein C-II in whole plasma. At concentrations of 0.012-0.06%, Tween-20 markedly increased the immunoreactivity of whole plasma, but not of delipidated plasma, indicating that antigenic sites of plasma apolipoprotein C-II has been exposed by Tween-20. In contrast, Triton X-100 had no effect on the immunoreactivity of whole plasma apolipoprotein C-II. A radioimmunoassay conducted in the presence of 0.06% Tween-20, resulted in a mean value in whole plasma (3.39 +/- 1.11 mg/dl) that was not significantly different from that obtained when the assay was done on delipidated samples. The immunoreactivity of VLDL apolipoprotein C-II was also drastically enhanced following lipolysis by bovine milk lipoprotein lipase, supporting the hypothesis that antigenic sites are masked by the lipids. Finally, the mechanism responsible for the effect of Tween-20 on apolipoprotein C-II immunoreactivity was investigated. The results obtained from circular dichroism and ultracentrifugation suggest that the detergent may dissociate the apolipoprotein from lipoprotein particles, thus fully exposing the antigenic sites for reaction with antibodies.