Development, High-Throughput Profiling, and Biopanning of a Large Phage Display Single-Domain Antibody Library

Overview

Journal Int J Mol Sci

Publisher MDPI

Specialties Biochemistry
Chemistry
Molecular Biology

Date 2024 May 11

PMID 38732011

Authors

Hee Eon Lee

Ah Hyun Cho

Jae Hyeon Hwang

Ji Woong Kim

Ha Rim Yang

Taehoon Ryu

Yushin Jung

Sukmook Lee

Affiliations

Soon will be listed here.

Abstract

Immunoglobulin G-based monoclonal antibodies (mAbs) have been effective in treating various diseases, but their large molecular size can limit their penetration of tissue and efficacy in multifactorial diseases, necessitating the exploration of alternative forms. In this study, we constructed a phage display library comprising single-domain antibodies (sdAbs; or "VHHs"), known for their small size and remarkable stability, using a total of 1.6 × 10 lymphocytes collected from 20 different alpacas, resulting in approximately 7.16 × 10 colonies. To assess the quality of the constructed library, next-generation sequencing-based high-throughput profiling was performed, analyzing approximately 5.65 × 10 full-length VHH sequences, revealing 92% uniqueness and confirming the library's diverse composition. Systematic characterization of the library revealed multiple sdAbs with high affinity for three therapeutically relevant antigens. In conclusion, our alpaca sdAb phage display library provides a versatile resource for diagnostics and therapeutics. Furthermore, the library's vast natural VHH antibody repertoire offers insights for generating humanized synthetic sdAb libraries, further advancing sdAb-based therapeutics.

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