RAPID-DETERMINATION OF APOLIPOPROTEIN-E PHENOTYPES FROM WHOLE PLASMA BY AUTOMATED ISOELECTRIC-FOCUSING USING PHASTSYSTEM(TM) AND IMMUNOFIXATION

The influence of the genetic apolipoprotein (apo) E isoforms on human plasma lipoproteins is well established. There is, however, still a ne ed for a phenotyping procedure applicable in laboratories not speciali zed in lipid research. To this end, we developed a rapid, automated el ectrophoresis method for apoE phenotyping. Either self-made or commerc ially available precasted gels can be used. Fresh or frozen samples co rresponding to 0.1 mu l of plasma are applied automatically after lipi d extraction in a urea-containing buffer onto the gel and isoelectric focusing is carried out for 45 min. Thereafter, apoE bands are precipi tated by specific polyclonal antibodies and visualized by automated si lver staining. The method is reliable, easily and quickly performed, a nd not restricted to specialized laboratories.