A single-domain antibody library based on a stability-engineered human VH3 scaffold.

Autor: Lee NJ; Department of Bioinspired Sciences, Ewha Womans University, Seoul, Korea., Jung M; Department of Bioinspired Sciences, Ewha Womans University, Seoul, Korea., Yang HY; Department of Life Sciences, Ewha Womans University, Seoul, Korea., Shim H; Department of Bioinspired Sciences, Ewha Womans University, Seoul, Korea. hshim@ewha.ac.kr.; Department of Life Sciences, Ewha Womans University, Seoul, Korea. hshim@ewha.ac.kr.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2024 Jul 31; Vol. 14 (1), pp. 17747. Date of Electronic Publication: 2024 Jul 31.
DOI: 10.1038/s41598-024-68680-5
Abstrakt: Using conventional immunoglobulin G (IgG) molecules as therapeutic agents presents several well-known disadvantages owing to their large size and structural complexity, negatively impacting development and production efficiency. Single-domain antibodies (sdAbs) are the smallest functional antibody format (~ 15 kDa) and represent a viable alternative to IgG in many applications. However, unlike natural single-domain antibodies, such as camelid V H H, the variable domains of conventional antibodies show poor physicochemical properties when expressed as sdAbs. This report identified stable sdAb variants of human VH3-23 from a framework region 2-randomized human VH library by phage display selection under thermal challenge. Synthetic complementarity determining region diversity was introduced to one of the selected variants with high thermal stability, expression level, and monomeric content to construct a human VH sdAb library. The library was validated by panning against a panel of antigens, and target-specific binders were identified and characterized for their affinity and biophysical properties. The results of this study suggest that a synthetic sdAb library based on a stability-engineered human VH scaffold could be a facile source of high-quality sdAb for many practical applications.
(© 2024. The Author(s).)
Databáze: MEDLINE
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