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用于酶联免疫吸附测定的抗体在尼龙上的固定化。

Immobilization of antibodies on nylon for use in enzyme-linked immunoassay.

作者信息

Hendry R M, Herrmann J E

出版信息

J Immunol Methods. 1980;35(3-4):285-96. doi: 10.1016/0022-1759(80)90255-0.

Abstract

Antibodies were immobilized by covalent linkage on nylon balls and powder for use in solid-phase enzyme-linked immunoassays. Covalent linkage of antibody to nylon was accomplished by treatment of partially hydrolyzed nylon with glutaraldehyde or carbodiimides. Up to 0.74 microgram of immunoglobulin G per mm2 nylon could be immobilized, whereas only 0.02 microgram per mm2 could be adsorbed to polystyrene, and the binding to nylon was stable. This eliminated the problem of antibody desorption noted in conventional enzyme-linked immunosorbent assay which are based on simple adsorption to plastics, and gave more reproducible results. The method was also more sensitive, detecting levels of approximately 1 ng per ml of immunoglobulin E in clinical samples. Further, antibodies coupled to nylon balls remained bound under conditions that dissociate antibody-antigen complexes, which permitted reuse of the immobilized antibodies for immunoassays.

摘要

抗体通过共价连接固定在尼龙球和粉末上,用于固相酶联免疫测定。抗体与尼龙的共价连接是通过用戊二醛或碳二亚胺处理部分水解的尼龙来实现的。每平方毫米尼龙最多可固定0.74微克免疫球蛋白G,而每平方毫米聚苯乙烯只能吸附0.02微克,并且与尼龙的结合是稳定的。这消除了传统酶联免疫吸附测定中基于简单吸附到塑料上所出现的抗体解吸问题,并给出了更可重复的结果。该方法也更灵敏,可检测临床样品中约每毫升1纳克的免疫球蛋白E水平。此外,与尼龙球偶联的抗体在使抗体 - 抗原复合物解离的条件下仍保持结合,这使得固定化抗体可重复用于免疫测定。

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