一种用于筛选杂交瘤上清液的改良斑点免疫结合试验。将未纯化的抗原固定在硝酸纤维素纸盘上。

An improved dot immunobinding assay for screening hybridoma supernatants. Non-purified antigen immobilized on nitrocellulose paper discs.

作者信息

Steinitz M, Rosén A, Klein G

机构信息

Department of Tumor Biology, Karolinska Institute, Stockholm, Sweden.

出版信息

J Immunol Methods. 1991 Jan 24;136(1):119-23. doi: 10.1016/0022-1759(91)90257-g.

DOI:10.1016/0022-1759(91)90257-g

PMID:1995705

Abstract

This report describes a modified dot immunobinding assay (DIA) in microplates using a crude mixture of non-purified antigen. Nitrocellulose filter paper discs exposed to the antigen mixture were inserted into the wells and kept in place by a specially constructed device. To test the efficiency of the modification a set of monoclonal antibodies from a mouse immunized with 58 kDa trpE-Bmyc fusion protein were screened. The advantage of this modified method over conventional ELISA is that it permits the use of non-purified antigen for screening large numbers of monoclonal antibodies.

摘要

本报告描述了一种在微孔板中使用未纯化抗原粗混合物的改良斑点免疫结合测定（DIA）。将暴露于抗原混合物的硝酸纤维素滤纸片插入孔中，并通过专门构建的装置固定在原位。为了测试改良的效率，筛选了一组用58 kDa trpE-Bmyc融合蛋白免疫的小鼠的单克隆抗体。这种改良方法相对于传统ELISA的优点在于，它允许使用未纯化的抗原筛选大量单克隆抗体。