Mandrusov E, Vroman L, Leonard E F
Department of Chemical Engineering, Material Science, and Mining Engineering, Columbia University, New York City, NY 10027, USA.
J Biomater Sci Polym Ed. 1996;8(1):1-18. doi: 10.1163/156856297x00542.
This paper describes and evaluates a method for quantifying the amounts of specific plasma proteins adsorbed to biomaterial surfaces. In particular, it demonstrates that macroscopic images ('stains'), that assess the spatial distribution of albumin, IgG, fibrinogen, and HMK (high molecular weight kininogen), can be obtained over areas of at least 12 cm2 using immunospecific adhesion of dyed polystyrene beads. Stain intensities, measured with a scanner and an image analysis system, were found to quantify the amount of specific protein in the solution used to coat the surfaces. Results obtained with the proposed method produced single protein isotherms for albumin, immunoglobulin G (IgG) and fibrinogen that followed Langmuir-like adsorption behavior and were similar to previously published isotherms. The HMK isotherm also exhibited Langmuir-like adsorption behavior. The proposed method also detected the presence of an expected maximum in the adsorption of fibrinogen onto glass as a function of plasma dilution. Adsorption of fibrinogen out of 6.4% plasma onto glass from a separated flow produced results indicating the quantity as well as the location of fibrinogen at the boundary of the separated region. This result confirmed the utility of the proposed method for detecting spatial distributions of specific proteins adsorbed from plasma in practical devices.
本文描述并评估了一种用于量化吸附在生物材料表面的特定血浆蛋白量的方法。具体而言，该方法表明，通过染色聚苯乙烯珠的免疫特异性黏附作用，能够在至少12平方厘米的区域上获取评估白蛋白、免疫球蛋白G（IgG）、纤维蛋白原和高分子量激肽原（HMK）空间分布的宏观图像（“染色图像”）。利用扫描仪和图像分析系统测量染色强度，结果发现其可量化用于包被表面的溶液中特定蛋白质的量。用该方法得到的结果产生了白蛋白、免疫球蛋白G（IgG）和纤维蛋白原的单蛋白等温线，这些等温线呈现出类似朗缪尔吸附的行为，且与先前发表的等温线相似。HMK等温线也表现出类似朗缪尔吸附的行为。该方法还检测到，作为血浆稀释度的函数，纤维蛋白原在玻璃上的吸附存在预期的最大值。从分流中6.4%的血浆中吸附到玻璃上的纤维蛋白原，其结果表明了在分离区域边界处纤维蛋白原的量以及位置。这一结果证实了该方法在检测实际装置中从血浆吸附的特定蛋白质空间分布方面的实用性。
