Measurement of small high density lipoprotein subclass by an improved immunoblotting technique.
An improved method of immunoblotting of plasma onto agarose gel matrix containing antiapolipoprotein A-I is described. Fresh plasma samples were subjected to gradient polyacrylamide gel electrophoresis (4-25%) followed by electrotransfer onto agarose gel layer containing antiapolipoprotein A-I. This method was compared with immunoblotting onto nitrocellulose where the transfer onto agarose gel matrix has been shown to be more convenient, quantitative, and can be kept permanently. Plasma apolipoprotein A-I was found to be distributed among regions of varying molecular weights ranging from 43,000 to 800,000. A small size fraction of molecular weight range of 43,000-50,000 (small HDL) was found in normolipidemic and hyperlipidemic subjects. The proportion of the latter fraction varied considerably among subjects (range: 0.0-32%), being lower in normolipidemic subjects (mean +/- SEM: 11.6 +/- 1.4%), and higher in hyperlipidemic subjects (mean +/- SEM: 23.7 %/- 1.7%, P < 0.001). Physiological increase in the level of the small HDL was observed in normolipidemic subjects 4 h after fat ingestion (difference: 5.0%, P < 0.001); moreover, the level was higher in normolipidemic subjects who consumed moderate amounts of alcohol (mean +/- SEM: 17.9 +/- 1.2%, P < 0.001) compared with normolipidemic subjects who do not drink alcohol at all.
Atmeh
R G
RG
Department of Chemical Sciences, University of Science and Technology, Irbred, Jordan.
Robenek
H
H
D000906:Antibodies; D016632:Apolipoprotein A-I / Q000276:immunology; D004041:Dietary Fats; D005260:Female; D006801:Humans; D007163:Immunosorbent Techniques; D008055:Lipids / Q000097:blood; D008075:Lipoproteins, HDL / Q000097:blood / Q000737:chemistry*; D008297:Male; D008970:Molecular Weight; D012685:Sepharose
D000906:Antibodies; D016632:Apolipoprotein A-I; D004041:Dietary Fats; D008055:Lipids; D008075:Lipoproteins, HDL; D012685:Sepharose