A HIGHLY SENSITIVE ASSAY FOR QUANTITATION OF APOLIPOPROTEIN B-48 USING AN ANTIBODY TO HUMAN APOLIPOPROTEIN-B AND ENHANCED CHEMILUMINESCENCE

We describe a method for the rapid quantification of serum apolipoprot ein B-48 using a commercially available anti-apolipoprotein (apo)B ant iserum and compare it to analytical sodium dodecyl sulphate polyacryla mide gel electrophoresis (SDS-PAGE) with coomassie blue R250 staining. The method described here eliminates the need for de-lipidation of sa mples and only requires a one-step overnight ultracentrifugation. West ern-blotting and enhanced chemiluminescence (ECL) visualization of pro teins was approximately 10 times more sensitive than coomassie stainin g and generally took no longer to complete than staining/ destaining o f SDS-PAGE gels. The sensitivity of the antiserum/ECL technique enable d quantitation of fasting apolipoprotein B-48 which could not be resol ved by SDS-PAGE and Coomassie staining.