Construction and validation of a synthetic phage-displayed nanobody library
10.4196/kjpp.2024.28.5.457
- Author:
Minju KIM
1
;
Xuelian BAI
;
Hyewon IM
;
Jisoo YANG
;
Youngju KIM
;
Minjoo MJ KIM
;
Yeonji OH
;
Yuna JEON
;
Hayoung KWON
;
Seunghyun LEE
;
Chang-Han LEE
Author Information
1. Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Korea
- Publication Type:Original Article
- From:The Korean Journal of Physiology and Pharmacology
2024;28(5):457-467
- CountryRepublic of Korea
- Language:English
-
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, phagedisplayed 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.
