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Yeast Mating as a Tool for Highly Effective Discovery and Engineering of Antibodies via Display Methodologies
  • Baek, Du San ;
  • Park, Seong Wook ;
  • Adams, Cynthia ;
  • Dimitrov, Dimiter S. ;
  • Kim, Yong Sung
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dc.contributor.authorBaek, Du San-
dc.contributor.authorPark, Seong Wook-
dc.contributor.authorAdams, Cynthia-
dc.contributor.authorDimitrov, Dimiter S.-
dc.contributor.authorKim, Yong Sung-
dc.date.issued2022-01-01-
dc.identifier.urihttps://aurora.ajou.ac.kr/handle/2018.oak/37044-
dc.identifier.urihttps://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=85129664763&origin=inward-
dc.description.abstractYeast surface display (YSD) is a powerful methodology for discovery and engineering of antibodies, and the yeast mating has been used to overcome low transformation efficiency of yeast in antibody library generation. We developed an optimized method of yeast mating for generating a large, combinatorial antibody fragment library and heterodimeric protein library by cellular fusion between two haploid cells carrying different library each other. This method allows for increased diversity in screening of target-specific fragment antigen-binding (Fab) antibodies as well as in the development of heterodimeric Fc variants for bi-specific antibody generation and T-cell receptor (TCR). Here we describe the efficient isolation of human antibodies against the activated GTP-bound form of the oncogenic Ras mutant (KRasG12D-GTP) by sequential isolation of their heavy chains (HCs) followed by combination with light chains (LCs) via the yeast mating process. This strategy facilitates guided selection of the antigen-specific HC with either a fixed functional LC, which has cytosol penetrating ability, or an LC library to generate the Fab. It also allows for deeper exploration of a sequence space with fixed diversity, leading to a higher probability of successful isolation of human antibodies with high specificity and affinity.-
dc.description.sponsorshipThis work was supported by a grant from the Priority Research Center Program (2019R1A6A1A11051471 to YSK) through the National Research Foundation of Korea (NRF), funded by the Ministry of Science, ICT & Future Planning, Republic of Korea, and a grant funded by Samsung Future Technology Center [grant number SRFC-MA1802-09 to YSK].-
dc.language.isoeng-
dc.publisherHumana Press Inc.-
dc.subject.meshAntibodies-
dc.subject.meshGuanosine Triphosphate-
dc.subject.meshHumans-
dc.subject.meshImmunoglobulin Fab Fragments-
dc.subject.meshPeptide Library-
dc.subject.meshSaccharomyces cerevisiae-
dc.titleYeast Mating as a Tool for Highly Effective Discovery and Engineering of Antibodies via Display Methodologies-
dc.typeBook Chapter-
dc.citation.endPage333-
dc.citation.startPage313-
dc.citation.titleMethods in Molecular Biology-
dc.citation.volume2491-
dc.identifier.bibliographicCitationMethods in Molecular Biology, Vol.2491, pp.313-333-
dc.identifier.doi10.1007/978-1-0716-2285-8_17-
dc.identifier.pmid35482198-
dc.identifier.scopusid2-s2.0-85129664763-
dc.identifier.urlhttp://www.springer.com/series/7651-
dc.subject.keywordAntibody library-
dc.subject.keywordAntibody screening-
dc.subject.keywordGuided selection-
dc.subject.keywordKRas-
dc.subject.keywordYeast mating-
dc.subject.keywordYeast surface display-
dc.type.otherBook Chapter-
dc.description.isoafalse-
dc.subject.subareaMolecular Biology-
dc.subject.subareaGenetics-
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