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Combinatorial Methods for Chemical and Biological Sensors pp 223–248Cite as

Combinatorial Libraries of Arrayable Single-Chain Antibodies

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Part of the book series: Integrated Analytical Systems ((ANASYS))

Abstract

Antibodies that bind their respective targets with high affinity and specificity have proven to be essential reagents for biological research. Antibody phage display has become the leading tool for the rapid isolation of single-chain variable fragment (scFv) antibodies in vitro for research applications, but there is usually a gap between scFv isolation and its application in an array format suitable for high-throughput proteomics. In this chapter, we present our antibody phage display system where antibody isolation and scFv immobilization are facilitated by the design of the phagemid vector used as platform. In our system, the scFvs are fused at their C-termini to a cellulose-binding domain (CBD) and can be immobilized onto cellulose-based filters. This made it possible to develop a unique filter lift screen that allowed the efficient screen for multiple binding specificities, and to directly apply library-derived scFvs in an antibody spotted microarray.

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Acknowledgments and Notes

The work reported in this chapter was supported in part by a research grant from the Israel Science Foundation, administered by the Israel National Academy for Sciences and Humanities (Jerusalem, Israel). The n-CoDeR® technology is covered by IP rights held by BioInvent, Sweden (http://www.bioinvent.com).

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  1. Department of Molecular Microbiology and Biotechnology, The George S. Wise Faculty of Life Sciences, Tel-Aviv University, Israel

    Itai Benhar

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  1. Itai Benhar
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  1. Global Research Center, General Electric Company, 12309, Niskayuna, NY, USA

    Radislav A. Potyrailo

  2. Nanobiotechnology, Lausitz University of Applied Sciences, 01968, Senftenberg, Germany

    Vladimir M. Mirsky

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Benhar, I. (2009). Combinatorial Libraries of Arrayable Single-Chain Antibodies. In: Potyrailo, R.A., Mirsky, V.M. (eds) Combinatorial Methods for Chemical and Biological Sensors. Integrated Analytical Systems. Springer, New York, NY. https://doi.org/10.1007/978-0-387-73713-3_9

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