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Oligovalent Fab Display on M13 Phage Improved by Directed Evolution

Vehniainen M; Sanmark H; Lamminmaki U; Huovinen T; Yla-Pelto J

dc.contributor.authorVehniainen M
dc.contributor.authorSanmark H
dc.contributor.authorLamminmaki U
dc.contributor.authorHuovinen T
dc.contributor.authorYla-Pelto J
dc.date.accessioned2022-10-27T11:54:20Z
dc.date.available2022-10-27T11:54:20Z
dc.identifier.urihttps://www.utupub.fi/handle/10024/155803
dc.description.abstractEfficient 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.
dc.language.isoen
dc.publisherHUMANA PRESS INC
dc.titleOligovalent Fab Display on M13 Phage Improved by Directed Evolution
dc.identifier.urnURN:NBN:fi-fe2021042714232
dc.relation.volume44
dc.contributor.organizationfi=PÄÄT Biotekniikka / FM|en=PÄÄT Biotechnology / FM|
dc.contributor.organization-code2606202
dc.converis.publication-id1640844
dc.converis.urlhttps://research.utu.fi/converis/portal/Publication/1640844
dc.format.pagerange221
dc.format.pagerange231
dc.identifier.jour-issn1073-6085
dc.okm.affiliatedauthorHuovinen, Tuomas
dc.okm.affiliatedauthorVehniäinen, Markus
dc.okm.affiliatedauthorSanmark, Hanna
dc.okm.affiliatedauthorLamminmäki, Urpo
dc.okm.discipline1182 Biochemistry, cell and molecular biologyen_GB
dc.okm.discipline1182 Biokemia, solu- ja molekyylibiologiafi_FI
dc.okm.internationalcopublicationnot an international co-publication
dc.okm.internationalityInternational publication
dc.okm.typeJournal article
dc.publisher.countryUnited Statesen_GB
dc.publisher.countryYhdysvallat (USA)fi_FI
dc.publisher.country-codeUS
dc.relation.doi10.1007/s12033-009-9231-3
dc.relation.ispartofjournalMolecular Biotechnology
dc.relation.issue3
dc.year.issued2010


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