用蛋白A进行表面修饰以实现单克隆抗体的均匀结合。

Surface modification with protein A for uniform binding of monoclonal antibodies.

作者信息

Schramm W, Yang T, Midgley A R

出版信息

Clin Chem. 1987 Aug;33(8):1338-42.

Abstract

We describe optimal conditions for immobilization of two monoclonal antibodies to progesterone for solid-phase assays. Polystyrene surfaces are refined with Protein A to achieve uniform, reproducible, stable, and sterically accessible immobilization of immunoglobulins (IgG). To this end, we optimized the amount of immobilized Protein A, the pH of the medium for immobilization, the concentration of antibody, and the polystyrene surface. We also investigated three carriers for solid-phase assays: 12 X 75 mm polystyrene test tubes, Macrowells (Skatron, Inc.; suitable for processing with multiple pipettors), and microwell strips (Immulon II, Dynatech Inc.). Immunoglobulin does not appreciably dissociate from any of these solid matrices, even if the assay procedure takes several hours. Therefore, we postulate that more than one molecule of immobilized Protein A binds to IgG, or that there is an additional interaction between the antibody and the polymer surface.

摘要

我们描述了将两种抗孕酮单克隆抗体固定用于固相测定的最佳条件。用蛋白A对聚苯乙烯表面进行处理，以实现免疫球蛋白（IgG）均匀、可重复、稳定且空间可及的固定。为此，我们优化了固定化蛋白A的用量、固定化介质的pH值、抗体浓度以及聚苯乙烯表面。我们还研究了三种用于固相测定的载体：12×75mm聚苯乙烯试管、微孔板（Skatron公司；适用于多道移液器操作）和微孔板条（Immulon II，Dynatech公司）。即使测定过程持续数小时，免疫球蛋白也不会从这些固体基质中明显解离。因此，我们推测固定化的蛋白A不止一个分子与IgG结合，或者抗体与聚合物表面之间存在额外的相互作用。