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Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries

1
Institute for Applied Biotechnology, Biberach University of Applied Science, Hubertus-Liebrecht-Strasse 35, 88400 Biberach, Germany
2
Faculty of Medicine, Graduate School “Molecular Medicine”, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany
3
Faculty of Natural Sciences, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany
*
Author to whom correspondence should be addressed.
Methods Protoc. 2019, 2(1), 17; https://doi.org/10.3390/mps2010017
Received: 28 December 2018 / Revised: 27 January 2019 / Accepted: 11 February 2019 / Published: 14 February 2019
Phage display is a powerful technique for drug discovery in biomedical research in particular for antibody libraries. But, several technical challenges are associated with the selection process. For instance, during the panning step, the successful elution of the phages bound to the antigen is critical in order to avoid losing the most promising binders. Here, we present an efficient protocol to establish, screen and select synthetic libraries of domain antibodies using phage display. We do not only present suitable solutions to the above-mentioned challenges to improve elution by 50-fold, but we also present a step by step in-depth protocol with miniaturized volumes and optimized procedures to save material, costs and time for a successful phage display with domain antibodies. Hence, this protocol improves the selection process for an efficient handling process. The here presented library is based on the variable domain (vNAR) of the naturally occurring novel antibody receptor (IgNAR) from cartilage fishes. Diversity was introduced in the Complementarity-Determining Region 3 (CDR3) of the antigen-binding site with different composition and length. View Full-Text
Keywords: display technology; antibody engineering; synthetic antibody library; shark antibody; vNAR; phage display; panning display technology; antibody engineering; synthetic antibody library; shark antibody; vNAR; phage display; panning
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MDPI and ACS Style

Solemani Zadeh, A.; Grässer, A.; Dinter, H.; Hermes, M.; Schindowski, K. Efficient Construction and Effective Screening of Synthetic Domain Antibody Libraries. Methods Protoc. 2019, 2, 17.

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