Open AccessHigh-yield expression of periplasmic single-chain variable fragments by solid Escherichia coli cultures
Author(s) -
Yoshiro Hanyu,
Macky Kato
Publication year - 2022
Publication title -
biotechniques/biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/btn-2021-0093
Subject(s) - periplasmic space , escherichia coli , recombinant dna , bacterial outer membrane , yield (engineering) , biology , liquid culture , microbiology and biotechnology , single chain variable fragment , chemistry , biochemistry , gene , physics , botany , thermodynamics
High-yield expression of quality antibody fragments is indispensable for research and diagnosis. Most recombinant antibody fragments are expressed in Escherichia coli using liquid cultures; however, their yields and quality are often poor. Here the authors expressed a single-chain variable fragment in E. coli cultivated on the wet surface of a solid support. Compared with a liquid culture, the authors obtained 2.5-times more single-chain variable fragments with membrane-cultivated E. coli. This method has two important advantages: it enables high yields of periplasmic single-chain variable fragments compared with liquid culture and offers simple and rapid expression and extraction.