Nimeri G, Lassen B, Gölander C G, Nilsson U, Elwing H
Department of Physics and Measurement Technology, Linköping University, Sweden.
J Biomater Sci Polym Ed. 1994;6(6):573-83. doi: 10.1163/156856294x00527.
Enzyme linked immunosorbent assay (ELISA) was used for the estimation of protein adsorption from blood plasma at some model solid surfaces. The majority of those surfaces were made in the wells of microtiterplates of polystyrene commonly used for ELISA purposes. Three of the model surfaces were made by radio frequency plasma discharge polymerization (RFPD) of the microtiterplates of polystyrene. The monomers we used were diaminocyclohexane, hexamethylenedisiloxane, and acrylic acid. Other surfaces investigated were: unmodified polystyrene, oxidized polystyrene, hydrophilic silicon oxide, and methylized silicon oxide. Two substances, Tween and bovine serum albumin (BSA), for the prevention of unintended adsorption of ELISA conjugate were also tested and the BSA method was found to be superior for this kind of investigation. Human blood plasma at different dilutions was incubated in the surface-modified microtiterplates followed by incubation of rabbit antibodies against fibrinogen (FG), fibronectin (FN), human serum albumin (HSA), complement factor 3 (C3), and immunoglobulin G (IgG). Visualization of bound antibodies was then made by standard ELISA procedure. At low blood plasma concentrations (plasma dil 1/1000), anti-IgG and anti-HSA were detected at high levels at the majority of surfaces. At high blood plasma concentration (plasma dil 1/10), anti-FG dominated at most surfaces. ELISA activity of FN and C3 were low at most of the surfaces at both plasma concentrations. An 'optimum' plasma dilution for the detection of surface bound FG (the Vroman effect) was not found with the use of the ELISA on any of the surfaces except for the silicon oxide surface. This is in contrast to findings by others who had used isotope-labelled fibrinogen diluted in plasma. However, 'false' Vroman effects occurred if nonionic surfactant was used for the prevention of unspecific binding in the ELISA.
酶联免疫吸附测定（ELISA）用于评估血浆中蛋白质在一些模型固体表面的吸附情况。这些表面大多是在常用于ELISA的聚苯乙烯微孔板孔中制成的。其中三个模型表面是通过对聚苯乙烯微孔板进行射频等离子体放电聚合（RFPD）制成的。我们使用的单体是二氨基环己烷、六甲基二硅氧烷和丙烯酸。其他研究的表面有：未改性的聚苯乙烯、氧化聚苯乙烯、亲水性氧化硅和甲基化氧化硅。还测试了两种用于防止ELISA缀合物意外吸附的物质，吐温（Tween）和牛血清白蛋白（BSA），发现BSA方法在这类研究中更具优势。将不同稀释度的人血浆在表面改性的微孔板中孵育，然后孵育抗纤维蛋白原（FG）、纤连蛋白（FN）、人血清白蛋白（HSA）、补体因子3（C3）和免疫球蛋白G（IgG）的兔抗体。然后通过标准ELISA程序对结合的抗体进行可视化检测。在低血浆浓度（血浆稀释1/1000）时，在大多数表面都能检测到高水平的抗IgG和抗HSA。在高血浆浓度（血浆稀释1/10）时，大多数表面以抗FG为主。在两种血浆浓度下，大多数表面的FN和C3的ELISA活性都很低。除氧化硅表面外，在任何表面上使用ELISA均未发现用于检测表面结合FG的“最佳”血浆稀释度（弗罗曼效应）。这与其他使用血浆中稀释的同位素标记纤维蛋白原的研究结果相反。然而，如果在ELISA中使用非离子表面活性剂来防止非特异性结合，则会出现“假”弗罗曼效应。
