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学科主题: 临床医学
题名:
Influences of amino acid sequences in FR1 region on binding activity of the scFv and Fab of an antibody to human gastric cancer cells
作者: Li, J; Wang, Y; Wang, ZZ; Dong, ZW
关键词: human gastric cancer ; scFv ; Fab ; PCR
刊名: IMMUNOLOGY LETTERS
发表日期: 2000-03-01
卷: 71, 期:3, 页:157-165
收录类别: SCI
文章类型: Article
WOS标题词: Science & Technology
类目[WOS]: Immunology
研究领域[WOS]: Immunology
关键词[WOS]: SINGLE-CHAIN FV ; ESCHERICHIA-COLI ; VARIABLE DOMAINS ; MONOCLONAL-ANTIBODIES ; SPECIFICITY ; AFFINITY ; EXPRESSION ; REPERTOIRE ; SECRETION ; RESIDUES
英文摘要:

Murine anti-gastric cancer mAb 3H11 has promising clinical applications. In this work, engineering of 3H11 scFv and Fab was conducted to increase its usefulness. 3H11 scFv and Fab were constructed by PCR amplification of the V-domains with degenerate primers for FR1. The bacterially expressed 3H11 Ab fragments showed no antigen binding activity. Then the N-terminal sequences of V regions were mutated to the 3H11 original sequence. The expressed scFv and Fab in bacterial culture supernatant showed binding activity to gastric cancer cells. Comparing the expression of unmutated and mutated 3H11 Fab, we found that the sequence changes of the V region N terminus introduced by PCR may seriously affect antigen binding but not the expression of antibody. Correction of either VL or VH N-terminal sequences can partially restore the antigen binding activity (61% for VL and 73% for VH). (C) 7000 Elsevier Science B.V. All rights reserved.

语种: 英语
WOS记录号: WOS:000086175500004
Citation statistics:
内容类型: 期刊论文
URI标识: http://ir.bjmu.edu.cn/handle/400002259/60673
Appears in Collections:北京大学临床肿瘤学院_期刊论文

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作者单位: 1.Beijing Med Univ, Sch Oncol, Beijing 100034, Peoples R China
2.Navy Gen Hosp, Beijing 100037, Peoples R China
3.Chinese Acad Med Sci, Inst Canc Res, Beijing 100021, Peoples R China

Recommended Citation:
Li, J,Wang, Y,Wang, ZZ,et al. Influences of amino acid sequences in FR1 region on binding activity of the scFv and Fab of an antibody to human gastric cancer cells[J]. IMMUNOLOGY LETTERS,2000,71(3):157-165.
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