A1 Refereed original research article in a scientific journal
Oligovalent Fab Display on M13 Phage Improved by Directed Evolution
Authors: Huovinen T, Sanmark H, Yla-Pelto J, Vehniainen M, Lamminmaki U
Publisher: HUMANA PRESS INC
Publication year: 2010
Journal: Molecular Biotechnology
Journal name in source: MOLECULAR BIOTECHNOLOGY
Journal acronym: MOL BIOTECHNOL
Number in series: 3
Volume: 44
Issue: 3
First page : 221
Last page: 231
Number of pages: 11
ISSN: 1073-6085
DOI: https://doi.org/10.1007/s12033-009-9231-3
Self-archived copy’s web address: https://research.utu.fi/converis/portal/Publication/1640844
Efficient display of antibody on filamentous phage M13 coat is crucial for successful biopanning selections. We applied a directed evolution strategy to improve the oligovalent display of a poorly behaving Fab fragment fused to phage gene-3 for minor coat protein (g3p). The Fab displaying clones were enriched from a randomly mutated Fab gene library with polyclonal anti-mouse IgG antibodies. Contribution of each mutation to the improved phenotype of one selected mutant was studied. It was found out that two point mutations had significant contribution to the display efficiency of Fab clones superinfected with hyperphage. The most dramatic effect was connected to a start codon mutation, from AUG to GUG, of the PelB signal sequence preceding the heavy chain. The clone carrying this mutation, FabM(GUG), displayed Fab 19-fold better and yielded twofold higher phage titers than the original Fab.
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