Codon-precise, synthetic, antibody fragment libraries built using automated hexamer codon additions and validated through next generation sequencing


We have previously described ProxiMAX, a technology that enables the fabrication of precise, combinatorial gene libraries via codon-by-codon saturation mutagenesis. ProxiMAX was originally performed using manual, enzymatic transfer of codons via blunt-end ligation. Here we present Colibra™: an automated, proprietary version of ProxiMAX used specifically for antibody library generation, in which double-codon hexamers are transferred during the saturation cycling process. The reduction in process complexity, resulting library quality and an unprecedented saturation of up to 24 contiguous codons are described. Utility of the method is demonstrated via fabrication of complementarity determining regions (CDR) in antibody fragment libraries and next generation sequencing (NGS) analysis of their quality and diversity.

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Divisions: College of Health & Life Sciences > School of Biosciences
College of Health & Life Sciences
College of Health & Life Sciences > School of Biosciences > Cellular and Molecular Biomedicine
Additional Information: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Uncontrolled Keywords: colibra,proxiMAX randomization,saturation mutagenesis,CDR,single domain antibody,camelid antibody,antibody engineering
Publication ISSN: 2073-4468
Last Modified: 05 Jan 2024 08:13
Date Deposited: 15 Jun 2015 12:05
Full Text Link: http://www.mdpi ... 073-4468/4/2/88
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PURE Output Type: Article
Published Date: 2015-05-15
Authors: Frigotto, Laura
Smith, Matthew E.
Brankin, Christopher
Sedani, Ashni
Cooper, Simon E.
Kanwar, Nisha
Evans, Daniel
Svobodova, Stanislava
Baar, Claudia
Glanville, Jacob
Ullman, Christopher G.
Hine, Anna V. (ORCID Profile 0000-0003-4065-831X)



Version: Published Version

License: Creative Commons Attribution

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