Affinity isoelectric focusing of human serum apolipoprotein B-containing lipoproteins on agarose gels with cyclodextrins.
In an attempt to subfractionate apolipoprotein B-containing lipoproteins (apoB-Lp), agarose isoelectric focusing (IEF) of apoB-Lp in human sera was carried out in the presence of two kinds of cyclodextrins (CDs), which have different affinities for cholesterol. ApoB-Lp in the agarose gels was detected by immunofixation, followed by protein staining. In the presence of both 11.4 mg/mL alpha-CD and 3.51 mg/mL beta-CD, but not either one alone, IEF separated a maximum of six apoB-Lp bands. Electrophoresis failed to separate the apoB-Lp subfractions under similar conditions with CDs. The patterns of apoB-Lp in IEF with the CDs appeared to differ in some sera. Since the subfractions of apoB-Lp can be separated without ultracentrifugation of serum, the present method serves for the screening of apoB-Lp heterogeneity on native serum lipoproteins.
Ichimura
M
M
School of Health Sciences, Medical School, Okayama University, Japan. miet@itan.okayama-u.ac.jp
Okamoto
M
M
Mori
S
S
Tohge
H
H
Ochi
K
K
Harada
H
H
D001055:Apolipoproteins B / Q000097:blood / Q000302:isolation & purification*; D003505:Cyclodextrins; D004587:Electrophoresis, Agar Gel / Q000379:methods*; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods*; D008074:Lipoproteins / Q000097:blood / Q000302:isolation & purification*; D047391:alpha-Cyclodextrins; D047392:beta-Cyclodextrins
D001055:Apolipoproteins B; D003505:Cyclodextrins; D008074:Lipoproteins; D047391:alpha-Cyclodextrins; D047392:beta-Cyclodextrins; C031215:betadex; C032613:alpha-cyclodextrin