从噬菌体展示组合文库中制备抗痘苗病毒的人单克隆抗体Fab片段并进行表征。
Production and characterization of human monoclonal antibody Fab fragments to vaccinia virus from a phage-display combinatorial library.
作者信息
Schmaljohn C, Cui Y, Kerby S, Pennock D, Spik K
机构信息
Virology Division, United States Army Medical Research Institute of Infectious Diseases, Frederick, Maryland, 21702-5011, USA.
出版信息
Virology. 1999 May 25;258(1):189-200. doi: 10.1006/viro.1999.9701.
Abstract
A combinatorial, phage-display library of human Fab antibody fragments was generated from IgG heavy chain (HC) and light chain (LC) genes cloned from the lymphocytes of a vaccinia virus (VACV)-immune donor. To ascertain the complexity of the library, nucleotide sequences of the variable regions of the HC and LC genes were determined. Fourteen distinct HC and 18 distinct LC (7 kappa and 11 lambda) that formed a combinatorial library of 22 Fabs were identified. Immune-precipitation of radiolabeled VACV revealed that at least six different VACV proteins were recognized by the antibodies. Plaque-reduction neutralization demonstrated that six of the Fabs neutralized VACV in the presence of anti-human antibody. ELISA studies indicated that 15 of the Fabs were cross-reactive with monkeypox virus.
摘要
从一名接种牛痘病毒（VACV）的免疫供体淋巴细胞中克隆出IgG重链（HC）和轻链（LC）基因，构建了一个组合噬菌体展示人Fab抗体片段文库。为确定文库的复杂性，测定了HC和LC基因可变区的核苷酸序列。鉴定出14种不同的HC和18种不同的LC（7种κ链和11种λ链），它们组成了一个包含22种Fab的组合文库。对放射性标记的VACV进行免疫沉淀显示，这些抗体至少识别六种不同的VACV蛋白。蚀斑减少中和试验表明，六种Fab在抗人抗体存在的情况下可中和VACV。ELISA研究表明，15种Fab与猴痘病毒具有交叉反应性。
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