Schaefer J R, Hackler R, Brand S, Schwarz S, Kleine T O, Steinmetz A
Abteilung Endokrinologie und Stoffwechsel, Universität Marburg, Germany.
Clin Chem. 1995 Jan;41(1):76-81.
Distinct genetic variants of apolipoprotein (apo) AI have been shown to influence concentrations of high-density lipoprotein (HDL) cholesterol. The genetic polymorphism of apo AIV may modulate HDL-cholesterol, plasma triglycerides, and lipoprotein(a) concentrations. There is evidence for an antagonizing role of apo AII in reverse cholesterol transport. Since genetic polymorphisms and variants of these apolipoproteins are detectable by isoelectric focusing (IEF), we developed a rapid and easy automated method for IEF analysis of apos AI, AII, and AIV on self-made or commercially available gels, using the PhastSystem. Diluted plasma or serum samples (1 microL) are applied automatically onto the gel and IEF is carried out for 35-45 min. Afterwards, the apo A bands are precipitated by specific polyclonal antibodies and visualized by automated silver staining. This rapid procedure is suitable as a routine or screening method for IEF analysis of these major HDL apolipoproteins.
载脂蛋白（apo）AI的不同基因变体已被证明会影响高密度脂蛋白（HDL）胆固醇的浓度。载脂蛋白AIV的基因多态性可能会调节HDL胆固醇、血浆甘油三酯和脂蛋白（a）的浓度。有证据表明载脂蛋白AII在逆向胆固醇转运中起拮抗作用。由于这些载脂蛋白的基因多态性和变体可通过等电聚焦（IEF）检测到，我们开发了一种快速简便的自动化方法，使用PhastSystem在自制或市售凝胶上对apo AI、AII和AIV进行IEF分析。将稀释的血浆或血清样品（1微升）自动加样到凝胶上，进行35 - 45分钟的IEF。之后，apo A条带通过特异性多克隆抗体沉淀，并通过自动银染进行可视化。这种快速方法适合作为这些主要HDL载脂蛋白IEF分析的常规或筛查方法。
