用SEREX法鉴定肿瘤相关自身抗原

Identification of tumor-associated autoantigens with SEREX.

作者信息

Türeci Ozlem, Usener Dirk, Schneider Sandra, Sahin Ugur

机构信息

Department of Internal Medicine, Johannes Gutenberg-University, Mainz, Germany.

出版信息

Methods Mol Med. 2005;109:137-54. doi: 10.1385/1-59259-862-5:137.

DOI:10.1385/1-59259-862-5:137

PMID:15585919

Abstract

Serological analysis of tumor antigens by recombinant cDNA expression cloning (SEREX) allows the systematic cloning of tumor antigens recognized by the spontaneous autoantibody repertoire of cancer patients. For SEREX, cDNA expression libraries are constructed from fresh tumor specimens, packaged into lambda-phage vectors, and expressed recombinantly in Escherichia coli. Recombinant proteins expressed during the lytic infection of bacteria are transferred onto nitrocellulose membranes to be probed with diluted autologous patient serum for identification of clones reactive with high-titered IgG antibodies. This chapter describes the SEREX technology in detail.

摘要

通过重组cDNA表达克隆技术对肿瘤抗原进行血清学分析（SEREX），能够系统地克隆癌症患者自发自身抗体库所识别的肿瘤抗原。对于SEREX而言，cDNA表达文库是由新鲜肿瘤标本构建而成，包装到λ噬菌体载体中，并在大肠杆菌中进行重组表达。在细菌裂解感染过程中表达的重组蛋白被转移到硝酸纤维素膜上，用稀释的自体患者血清进行检测，以鉴定与高滴度IgG抗体反应的克隆。本章将详细介绍SEREX技术。