噬菌体展示技术在高通量抗体生成与表征中的应用。

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

Abcam Ltd, Cambridge Science Park, Cambridge CB4 0FW, UK.

Genome Biol. 2007;8(11):R254. doi: 10.1186/gb-2007-8-11-r254.

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.

我们构建了一个高质量的噬菌体展示文库，其中包含超过10¹⁰种人源抗体，并描述了其在前所未有的规模上用于产生抗体的情况。我们已对292种抗原筛选、测序了超过38,000种重组抗体，获得了超过7,200个独特克隆。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，相关数据/克隆可在线获取。在基于微珠的流式细胞术检测中证明了其灵敏检测能力。此外，在组织微阵列上通过免疫组织化学发现37%（143/381）的抗体呈阳性染色。因此，我们展示了全基因组单克隆抗体产生的潜力，并阐明了与之相关的问题。