The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
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Construction and validation of a synthetic phage-displayed nanobody library

  • Minju Kim (Department of Biomedical Sciences, Seoul National University College of Medicine) ;
  • Xuelian Bai (Research Center, EPD Biotherapeutics Inc.) ;
  • Hyewon Im (Cancer Research Institute) ;
  • Jisoo Yang (Department of Biomedical Sciences, Seoul National University College of Medicine) ;
  • Youngju Kim (Department of Biomedical Sciences, Seoul National University College of Medicine) ;
  • Minjoo MJ Kim (Department of Biomedical Sciences, Seoul National University College of Medicine) ;
  • Yeonji Oh (Research Center, EPD Biotherapeutics Inc.) ;
  • Yuna Jeon (Research Center, EPD Biotherapeutics Inc.) ;
  • Hayoung Kwon (Department of Pharmacology, Seoul National University College of Medicine) ;
  • Seunghyun Lee (Research Center, EPD Biotherapeutics Inc.) ;
  • Chang-Han Lee (Department of Biomedical Sciences, Seoul National University College of Medicine)
  • Received : 2024.04.10
  • Accepted : 2024.04.22
  • Published : 2024.09.01
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Abstract

Nanobodies derived from camelids and sharks offer unique advantages in therapeutic applications due to their ability to bind to epitopes that were previously inaccessible. Traditional methods of nanobody development face challenges such as ethical concerns and antigen toxicity. Our study presents a synthetic, phage-displayed nanobody library using trinucleotide-directed mutagenesis technology, which allows precise amino acid composition in complementarity-determining regions (CDRs), with a focus on CDR3 diversity. This approach avoids common problems such as frameshift mutations and stop codon insertions associated with other synthetic antibody library construction methods. By analyzing FDA-approved nanobodies and Protein Data Bank sequences, we designed sub-libraries with different CDR3 lengths and introduced amino acid substitutions to improve solubility. The validation of our library through the successful isolation of nanobodies against targets such as PD-1, ATXN1 and STAT3 demonstrates a versatile and ethical platform for the development of high specificity and affinity nanobodies and represents a significant advance in biotechnology.

Keywords

Acknowledgement

This work was supported by grant no 03-2020-0300 from the SNUH Research Fund and the Creative-Pioneering Researchers Program through Seoul National University.

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