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Int. J. Mol. Sci. 2017, 18(6), 1169; doi:10.3390/ijms18061169

Utilization of Multi-Immunization and Multiple Selection Strategies for Isolation of Hapten-Specific Antibodies from Recombinant Antibody Phage Display Libraries

VTT Technical Research Centre of Finland, Tietotie 2, FI-02150 Espoo, Finland
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Academic Editor: Li Lin
Received: 3 May 2017 / Revised: 24 May 2017 / Accepted: 26 May 2017 / Published: 31 May 2017
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

Phage display technology provides a powerful tool for the development of novel recombinant antibodies. In this work, we optimized and streamlined the recombinant antibody discovery process for haptens as an example. A multi-immunization approach was used in order to avoid the need for construction of multiple antibody libraries. Selection methods were developed to utilize the full potential of the recombinant antibody library by applying four different elution conditions simultaneously. High-throughput immunoassays were used to analyse the binding properties of the individual antibody clones. Different carrier proteins were used in the immunization, selection, and screening phases to avoid enrichment of the antibodies for the carrier protein epitopes. Novel recombinant antibodies against mycophenolic acid and ochratoxin A, with affinities up to 39 nM and 34 nM, respectively, were isolated from a multi-immunized fragment antigen-binding (Fab) library. View Full-Text
Keywords: phage display; fragment antigen-binding (Fab); recombinant antibody; high-throughput; hapten; mycotoxin phage display; fragment antigen-binding (Fab); recombinant antibody; high-throughput; hapten; mycotoxin
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Tullila, A.; Nevanen, T.K. Utilization of Multi-Immunization and Multiple Selection Strategies for Isolation of Hapten-Specific Antibodies from Recombinant Antibody Phage Display Libraries. Int. J. Mol. Sci. 2017, 18, 1169.

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