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Updated: Aug 16, 2026
Scalable High Throughput Selection From Phage-displayed Synthetic Antibody Libraries
Published on: January 17, 2015
In vitro selection of VNAR antibodies using bioinformatics-based library design and cDNA display
Hiroki Anzai1, Takuya Terai2, Susumu Mitsuyama1,3
1Laboratory of Aquatic Molecular Biology and Biotechnology, Department of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
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Variable domains of new antigen receptors (VNARs) derived from shark immunoglobulin new antigen receptors (IgNARs) have gained attention as promising single-domain antibody modalities owing to their compact structures, structural stability, and unique binding capabilities. This study aimed to demonstrate the feasibility of screening a synthetic library based on VNARs derived from the brownbanded bamboo shark (Chiloscyllium punctatum) using a complementary DNA (cDNA) display system. A synthetic VNAR library was constructed by randomizing the CDR1 and CDR3 loops, resulting in a genetic diversity of approximately 1012 sequences in the cDNA display format. Using fluorescent proteins as model antigens, in vitro selection was performed, followed by next-generation sequencing and biochemical validation. VNAR-E1 and VNAR-N1 were identified as binding to EGFP and mNeonGreen, respectively. This work represents the first demonstration of applying a cDNA display system to VNAR selection.
