通过噬菌体展示产生的抗β2糖蛋白I抗体和抗凝血酶原抗体。
Anti-beta(2)GP-I and anti-prothrombin antibodies generated by phage display.
作者信息
Cabiedes Javier, Cabral Antonio R
机构信息
Department of Immunology and Rheumatology, Instituto Nacional de Ciencias Médicas y Nutrición Salvador Zubirán, Tlalpan, Mexico City, 14000, Mexico.
出版信息
Methods Mol Biol. 2009;562:61-70. doi: 10.1007/978-1-60327-302-2_5.
This chapter describes the construction and screening of a library of single-chain variable fragments (svFv) derived from patients with autoimmune disease. The methods cover the isolation of mononuclear cells from peripheral blood, preparation of RNA, and recovery of immunoglobulin-coding sequences by polymerase chain reaction (PCR). Cloning into a phage display vector and screening of the scFv display library by a simple panning procedure are described. These methods are applicable to library construction from any patient group or (with alternative primer sets) any mammalian species.
本章描述了从自身免疫性疾病患者中获得的单链可变片段（scFv）文库的构建和筛选方法。这些方法包括从外周血中分离单核细胞、制备RNA以及通过聚合酶链反应（PCR）回收免疫球蛋白编码序列。还介绍了将其克隆到噬菌体展示载体中，并通过简单的淘选程序筛选scFv展示文库的方法。这些方法适用于从任何患者群体构建文库，或（使用替代引物组）从任何哺乳动物物种构建文库。
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