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抗体固定到辉光放电处理过的聚合物上。

Antibody immobilization onto glow discharge treated polymers.

作者信息

Safranj A, Kiaei D, Hoffman A S

机构信息

Center for Bioengineering, University of Washington, Seattle 98195.

出版信息

Biotechnol Prog. 1991 Mar-Apr;7(2):173-7. doi: 10.1021/bp00008a012.

Abstract

Previous studies have shown that certain glow discharge treated polymers strongly retain adsorbed albumin and fibrinogen. On the basis of this phenomenon, we have investigated the possibility of immobilizing antibodies on glow discharge treated surfaces for diagnostic immunoassay applications. As a model for antibody immobilization, bovine IgG was immobilized on the following polymers: polyethylene (PE), tetrafluoroethylene glow discharge treated PE (TFE/PE), poly(ethylene terephthalate) (PET), TFE/PET, poly(tetrafluoroethylene) (PTFE), ethylene glow discharge treated PET (E/PET) and hexamethyldisiloxane glow discharge treated PET (HMDS/PET). IgG was radiolabeled with 125I and immobilized by either of the following two methods: (a) physical adsorption of IgG on untreated and glow discharge treated polymers or (b) physical adsorption of albumin followed by chemical coupling of IgG to albumin by glutaraldehyde. IgG concentration as well as adsorption times were varied in order both to optimize the immobilization conditions and to investigate the adsorption and retention mechanisms. To evaluate the efficiency of the immobilization techniques, blood plasma, Tween-20, and sodium dodecyl sulfate (SDS) were used to elute the adsorbed IgG layer. We found that IgG was successfully immobilized on the fluorocarbon glow discharge treated surfaces by using either the physical adsorption or the glutaraldehyde coupling method, although the former is more efficient than the latter method.

摘要

先前的研究表明,某些经辉光放电处理的聚合物能强烈保留吸附的白蛋白和纤维蛋白原。基于这一现象,我们研究了在辉光放电处理过的表面固定抗体以用于诊断免疫分析应用的可能性。作为抗体固定的模型,牛免疫球蛋白G(bovine IgG)被固定在以下聚合物上:聚乙烯(PE)、经四氟乙烯辉光放电处理的PE(TFE/PE)、聚对苯二甲酸乙二酯(PET)、TFE/PET、聚四氟乙烯(PTFE)、经乙烯辉光放电处理的PET(E/PET)以及经六甲基二硅氧烷辉光放电处理的PET(HMDS/PET)。IgG用125I进行放射性标记,并通过以下两种方法之一进行固定:(a)将IgG物理吸附在未处理和经辉光放电处理的聚合物上,或(b)先物理吸附白蛋白,然后通过戊二醛将IgG与白蛋白进行化学偶联。改变IgG浓度以及吸附时间,以便优化固定条件并研究吸附和保留机制。为了评估固定技术的效率,使用血浆、吐温-20和十二烷基硫酸钠(SDS)来洗脱吸附的IgG层。我们发现,通过物理吸附或戊二醛偶联方法,IgG都成功固定在了经氟碳辉光放电处理的表面上,尽管前者比后者更有效。

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