Bale M D, Wohlfahrt L A, Mosher D F, Tomasini B, Sutton R C
Life Sciences Research Laboratories, Eastman Kodak Company, Rochester, NY 14650-2113.
Blood. 1989 Dec;74(8):2698-706.
The arrays of proteins adsorbed from plasma onto a series of polystyrene copolymeric latexes were analyzed by enzyme-linked immunosorbent assay (ELISA) of washed beads and immunoblotting of proteins desorbed from the beads and separated by polyacrylamide gel electrophoresis (PAGE). Beads were prepared by continuous emulsion polymerization in the absence of surfactant. Coomassie brilliant blue staining of gel electropherograms of desorbed proteins indicated that the presence of small amounts of comonomers (1 to 10 mole %) significantly influenced the composition of the adsorbed protein layer. Immunoblotting revealed that fibrinogen, fibronectin, and vitronectin were adsorbed by all surfaces investigated. C3 and Clq adsorption varied significantly with copolymer composition. The ELISAs revealed that although the concentrations of vitronectin and fibronectin in plasma are similar, the extent of vitronectin adsorption from 70% to 85% plasma was greater by two orders of magnitude than fibronectin adsorption. Vitronectin adsorbed on carboxylic acid-containing copolymers reacted more strongly with a conformationally sensitive antivitronectin monoclonal antibody (MoAb) than vitronectin adsorbed to polystyrene and was more susceptible to cleavage by plasma proteases(s). The results show that vitronectin is a major protein adsorbed from concentrated plasma and that small changes in the chemical composition of a copolymer profoundly affects the extent and nature of protein adsorption from complex mixtures such as plasma.
通过对洗涤后的珠子进行酶联免疫吸附测定（ELISA）以及对从珠子上解吸并经聚丙烯酰胺凝胶电泳（PAGE）分离的蛋白质进行免疫印迹分析，研究了从血浆吸附到一系列聚苯乙烯共聚乳胶上的蛋白质阵列。珠子是在无表面活性剂的情况下通过连续乳液聚合制备的。对解吸蛋白质的凝胶电泳图谱进行考马斯亮蓝染色表明，少量共聚单体（1至10摩尔%）的存在显著影响吸附蛋白层的组成。免疫印迹显示，纤维蛋白原、纤连蛋白和玻连蛋白被所有研究的表面吸附。C3和Clq的吸附随共聚物组成有显著变化。ELISA结果表明，尽管血浆中玻连蛋白和纤连蛋白的浓度相似，但在70%至85%血浆中玻连蛋白的吸附程度比纤连蛋白的吸附程度大两个数量级。吸附在含羧酸共聚物上的玻连蛋白与构象敏感的抗玻连蛋白单克隆抗体（MoAb）的反应比吸附在聚苯乙烯上的玻连蛋白更强，并且更容易被血浆蛋白酶切割。结果表明，玻连蛋白是从浓缩血浆中吸附的主要蛋白质，共聚物化学成分的微小变化会深刻影响从血浆等复杂混合物中蛋白质吸附的程度和性质。
