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Antibody arrays for high-throughput screening of antibody–antigen interactions

Nature Biotechnology volume 18, pages 989–994 (2000)Cite this article

Abstract

We have developed a novel technique for high-throughput screening of recombinant antibodies, based on the creation of antibody arrays. Our method uses robotic picking and high-density gridding of bacteria containing antibody genes followed by filter-based enzyme-linked immunosorbent assay (ELISA) screening to identify clones that express binding antibody fragments. By eliminating the need for liquid handling, we can thereby screen up to 18,342 different antibody clones at a time and, because the clones are arrayed from master stocks, the same antibodies can be double spotted and screened simultaneously against 15 different antigens. We have used our technique in several different applications, including isolating antibodies against impure proteins and complex antigens, where several rounds of phage display often fail. Our results indicate that antibody arrays can be used to identify differentially expressed proteins.

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Figure 1: Antibody array screening after one round of phage selection using BSA.
Figure 2: Antibody array screening after one round of phage selection using a mixture of three unpurified recombinant bacterial lysates.
Figure 3: Western blot analysis of HeLa cells probed with scFv clones 2M15, 2G11, 8F20, 14H10, and 20M8 and Con (a non-binding scFv control).

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Acknowledgements

We thank the RZPD Resource centre (Berlin, Germany) for providing clones from the human fetal brain cDNA library hEX1, and Lucy Holt for help during experimental work and comments on the manuscript.

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Authors and Affiliations

  1. MRC Laboratory of Molecular Biology and MRC Centre for Protein Engineering, Hills Road, Cambridge, CB2 2QH, UK

    Ruud M. T. de Wildt & Ian M. Tomlinson

  2. UK Human Genome Mapping Project Resource Centre, Hinxton Hall, Hinxton, Cambridge, CB10 1SB, UK

    Chris R. Mundy & Barbara D. Gorick

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  1. Ruud M. T. de Wildt
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  2. Chris R. Mundy
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  4. Ian M. Tomlinson
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Correspondence to Ruud M. T. de Wildt or Ian M. Tomlinson.

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de Wildt, R., Mundy, C., Gorick, B. et al. Antibody arrays for high-throughput screening of antibody–antigen interactions. Nat Biotechnol 18, 989–994 (2000). https://doi.org/10.1038/79494

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