A Highly Sensitive Assay for Quantitation of Apolipoprotein B 48 Using an Antibody to Human Apolipoprotein B and Enhanced Chemiluminescence

Abstract

We describe a method for the rapid quantification of serum apolipoprotein B₄₈ using a commercially available anti-apolipoprotein (apo)B antiserum and compare it to analytical sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) with coomassie blue R250 staining. The method described here eliminates the need for de-lipidation of samples and only requires a one-step overnight ultracentrifugation. Western-blotting and enhanced chemiluminescence (ECL) visualization of proteins was approximately 10 times more sensitive than coomassie staining and generally took no longer to complete than staining/destaining of SDS-PAGE gels. The sensitivity of the antiserum/ECL technique enabled quantitation of fasting apolipoprotein B₄₈ which could not be resolved by SDS-PAGE and Coomassie staining.

Keywords

hypercholesterolaemia LDL triglyceride-rich lipoprotein

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