Ebato H, Gentry C A, Herron J N, Müller W, Okahata Y, Ringsdorf H, Suci P A
Department of Polymer Chemistry, Tokyo Institute of Technology, Japan.
Anal Chem. 1994 May 15;66(10):1683-9. doi: 10.1021/ac00082a014.
Antifluorescyl IgG antibody and Fab binding to two fluorescein-conjugated lipids was measured using the quartz crystal microbalance methodology. By use of the Langmuir-Blodgett technique, the fluorescein lipids, which were diluted to 5% in a L-alpha-dipalmitoyl phosphatidylethanolamine (DPPE) matrix, were deposited directly onto one gold electrode of the quartz crystal. Binding to films containing the fluorescein hapten was significantly enhanced compared to films of the pure DPPE matrix lipid, indicating that binding occurred primarily through a specific interaction. Association constants were 40-300 times less than for binding to haptens free in solution. Binding of IgG to the lipid in which the hydrocarbon chains and the fluorescein hapten were linked via a hydrophilic spacer was approximately 7 times as great as to the lipid containing no spacer. IgG binding to the lipid containing the spacer was increased 1.5-4.4 times compared to Fab binding for the same lipid. Equilibrium binding curves and kinetic measurements are analyzed quantitatively and compared.
使用石英晶体微天平方法测量抗荧光素IgG抗体和Fab与两种荧光素偶联脂质的结合。通过朗缪尔-布洛杰特技术，将在L-α-二棕榈酰磷脂酰乙醇胺（DPPE）基质中稀释至5%的荧光素脂质直接沉积在石英晶体的一个金电极上。与纯DPPE基质脂质膜相比，与含有荧光素半抗原的膜的结合显著增强，表明结合主要通过特异性相互作用发生。缔合常数比与溶液中游离半抗原的结合小40-300倍。IgG与其中烃链和荧光素半抗原通过亲水间隔基连接的脂质的结合约为与不含间隔基的脂质结合的7倍。与相同脂质的Fab结合相比，IgG与含间隔基脂质的结合增加了1.5-4.4倍。对平衡结合曲线和动力学测量进行了定量分析和比较。
