Application of phage display to high throughput antibody generation and characterization.
Application of phage display to high throughput antibody generation and characterization.
复制标题
将噬菌体显示到高吞吐抗体的产生和表征。
DOI：
10.1186/gb-2007-8-11-r254
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发表时间：
2007
期刊：
影响因子：
12.3
通讯作者：
McCafferty, John
中科院分区：
文献类型：
--
作者：
Schofield, Darren J;Pope, Anthony R;Clementel, Veronica;Buckell, Jenny;Chapple, Susan Dj;Clarke, Kay F;Conquer, Jennie S;Crofts, Anna M;Crowther, Sandra R E;Dyson, Michael R;Flack, Gillian;Griffin, Gareth J;Hooks, Yvette;Howat, William J;Kolb-Kokocinski, Anja;Kunze, Susan;Martin, Cecile D;Maslen, Gareth L;Mitchell, Joanne N;O'Sullivan, Maureen;Perera, Rajika L;Roake, Wendy;Shadbolt, S Paul;Vincent, Karen J;Warford, Anthony;Wilson, Wendy E;Xie, Jane;Young, Joyce L;McCafferty, John
已经构建了含有超过1010种人抗体的噬菌体展示文库，允许大规模产生抗体。选择、筛选和测序了针对292种抗原的38，000多个重组抗体，并进一步表征了4，400个所得独特克隆。我们已经创建了一个高质量的噬菌体展示库，其中包含超过1010种人抗体，并描述了其在前所未有的规模上产生抗体的用途。我们已经选择，筛选和测序了292种抗原的38，000多个重组抗体，产生了7，200多个独特的克隆。通过特异性测试和详细的序列分析对4，400种抗体进行了表征，数据/克隆可在线获得。在基于珠的流式细胞术测定中证明了灵敏的检测。此外，通过免疫组织化学在组织微阵列上发现37%（143/381）的抗体阳性染色。因此，我们已经证明了与全基因组单克隆抗体产生相关的问题的潜力和照明。
A phage display library has been constructed containing over 1010 human antibodies, allowing the large-scale generation of antibodies. Over 38,000 recombinant antibodies against 292 antigens were selected, screened and sequenced, and 4,400 resultant unique clones characterized further. 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.