载脂蛋白免疫复合物微粒的共振散射光谱研究及分析应用

在pH6.8的Na2HPO4-NaH2PO4缓冲溶液中和聚乙二醇存在下,载脂蛋白A1(APOA1)、载脂蛋白B(APOB)与相应的抗体发生特异性结合,分别形成粒径约为180,140nm的抗体抗原免疫复合物微粒,导致体系在340,470nm处的共振散射峰增强。分别研究了pH、抗血清、聚乙二醇浓度、温育时间和共存物质的影响。在最佳条件下,APOA1浓度在8.4~430.0ng·mL-1,APOB浓度在14.8~590.0ng·mL-1范围内与其共振散射强度成良好线性关系,方法的检出限分别为6.2和7.0ng·mL-1,用于定量分析血清中的APOA1与APOB,获得满意结果。

Resonance Scattering Spectra of Apolipoprotein Immunocomplex Particles and Its Analytical Application

In pH 6.8 Na2 HPO4-NaH2PO4 buffer solution and in presence of polyethylene glycol (PEG), apolipoprotein AI (ApoAI) and apolipoprotein B (ApoB) would combine with their corresponding antibody and produced immune complex particles in size of about 180 nm and 140 nm respectively, which exhibit stronger resonance scattering (RS) effect at 340 nm and 470 nm. The influence of pH, antisera volume, PEG concentration, incubation time and co-exists substances was considered. Under the optimal conditions, the RS intensity is proportional to the concentrations of ApoAI and ApoB when their concentration is in the range of 8.4-430.0 ng x mL(-1) and 14.8-590.0 ng x mL(-1), respectively. The detection limits (DL) are 6.2 ng x mL(-1) for ApoAI and 7.0 ng x mL(-1) for ApoB. The method was successfully applied to determination of ApoAI and ApoB in human serum samples, with satisfactory results.