Application of phage display to high throughput antibody generation and characterization

被引:166
|
作者
Schofield, Darren J. [2 ]
Pope, Anthony R. [3 ]
Clementel, Veronica [4 ]
Buckell, Jenny [5 ]
Chapple, Susan D. J. [3 ]
Clarke, Kay F. [3 ]
Conquer, Jennie S. [6 ]
Crofts, Anna M. [3 ]
Crowther, Sandra R. E. [3 ]
Dyson, Michael R. [1 ]
Flack, Gillian [7 ]
Griffin, Gareth J. [3 ]
Hooks, Yvette [3 ]
Howat, William J. [8 ]
Kolb-Kokocinski, Anja [3 ]
Kunze, Susan [4 ]
Martin, Cecile D. [9 ]
Maslen, Gareth L. [3 ]
Mitchell, Joanne N. [8 ]
O'Sullivan, Maureen [10 ]
Perera, Rajika L. [9 ]
Roake, Wendy [3 ]
Shadbolt, S. Paul [3 ]
Vincent, Karen J. [11 ]
Warford, Anthony [12 ]
Wilson, Wendy E. [3 ]
Xie, Jane [3 ]
Young, Joyce L. [4 ]
McCafferty, John [1 ]
机构
[1] Univ Cambridge, Dept Biochem, Cambridge CB2 1QW, England
[2] Abcam Ltd, Cambridge CB4 0FW, England
[3] Wellcome Trust Sanger Inst, Hinxton CB10 1HH, Cambs, England
[4] Cambridge Antibody Technol, Cambridge CB21 6GH, England
[5] Canc Res UK, Res Monoclonal Antibody Serv, London WC2A 3PX, England
[6] Mol Products Ltd, Thanxted CM6 2LT, England
[7] Bedford Hosp NHS Trust, Cell Histopathol Dept, Bedford MK42 9DJ, England
[8] Canc Res UK, Cambridge Res Inst, Cambridge CB2 0RE, England
[9] Glaxo Smith Kline Med Res Ctr, Stevenage SG1 2NY, Herts, England
[10] Genzyne Therapeut Ltd, Cambridge CB4 0WG, England
[11] Novartis Inst Biomed Res, Discov Technol, CH-4002 Basel, Switzerland
[12] Astra Zeneca Innovat Ctr China, Shanghai 200041, Peoples R China
基金
英国惠康基金;
关键词
D O I
10.1186/gb-2007-8-11-r254
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
We have created a high quality phage display library containing over 1010 human antibodies and describe its use in the generation of antibodies on an unprecedented scale. We have selected, screened and sequenced over 38,000 recombinant antibodies to 292 antigens, yielding over 7,200 unique clones. 4,400 antibodies were characterized by specificity testing and detailed sequence analysis and the data/clones are available online. Sensitive detection was demonstrated in a bead based flow cytometry assay. Furthermore, positive staining by immunohistochemistry on tissue microarrays was found for 37% (143/381) of antibodies. Thus, we have demonstrated the potential of and illuminated the issues associated with genome-wide monoclonal antibody generation.
引用
收藏
页数:18
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