通过等电聚焦和将血清抗体库转移至硝酸纤维素纸上进行毛细管印迹分析。

Analysis of serum antibody repertoires by isoelectric focusing and capillary blotting onto nitrocellulose paper.

作者信息

Schibeci A, Wade A W, Depew W T, Szewczuk M R

出版信息

J Immunol Methods. 1986 May 22;89(2):201-5. doi: 10.1016/0022-1759(86)90358-3.

Abstract

A rapid method for the analysis of antigen-specific human serum antibodies is described. After isoelectric focusing on thin agarose gels, serum proteins are transferred to nitrocellulose paper by capillary blotting. The kinetics of transfer indicated that ca. 90% of 125I-labelled isolated human immunoglobulins (IgG, IgA and IgM) are transferred to the nitrocellulose paper within 10 min. Probing of the nitrocellulose blot with 125I-labelled tetanus toxoid antigen followed by autoradiography reveals the total serum anti-tetanus toxoid antibody repertoire, including high molecular weight IgM. This procedure is rapid (results can be obtained in less than 12 h), sensitive and should prove very useful for detailed studies and analysis of antibody repertoires in body fluids and extracts

摘要

本文描述了一种分析抗原特异性人血清抗体的快速方法。在薄琼脂糖凝胶上进行等电聚焦后，血清蛋白通过毛细管印迹转移到硝酸纤维素纸上。转移动力学表明，约90%的125I标记的分离人免疫球蛋白（IgG、IgA和IgM）在10分钟内转移到硝酸纤维素纸上。用125I标记的破伤风类毒素抗原探测硝酸纤维素印迹，然后进行放射自显影，可揭示血清中总的抗破伤风类毒素抗体库，包括高分子量IgM。该方法快速（不到12小时即可获得结果）、灵敏，对于体液和提取物中抗体库的详细研究和分析将非常有用。