Abstract
The procedure for the measurement of serum apoprotein E by rocket immunoelectrophoresis is described. Very low density lipoprotein (VLDL) was isolated from fresh pooled human plasma by ultracentrifugation. The VLDL was delipidated with tetramethylurea. This material was underwent preparative SDS polyacrylamide gel electrophoresis and the protein bands were visualized by placing gels into 4 M sodium acetate. Regions of the gels conteining apoprotein E were cut from the gels and pooled. Apo E was eluted from the gels with SDS-Tris buffer. The antisera against apo E was produced by immunizing rabbits. Rocket immunoelectrophoresis for quantitative determination of human serum apo E was performed by using 1% agarose gel plates containing 2% (v/v) antisera.
Serum apoprotein E levels of hyperlipidemic subjects were markedly elevated and correlated with serum triglyceride concentrations but not with cholesterol concentrations. In liver diseases, levels of serum apo E were increased in patients with acute hepatitis, chronic hepatitis, compensated liver cirrhosis and obstructive jaundice. In contrast, patients with decompensated liver cirrhosis had decreased serum apoprotein E levels. Serum apoprotein E levels in patients with liver disease were correlated with both serum triglyceride and cholesterol concentrations.
