Development of an in vitro screening system for synthetic signal peptide in mammalian cell?based protein production

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dc.contributor.authorJong-Ho Park-
dc.contributor.authorHoon Min Lee-
dc.contributor.authorEun-Ju Jin-
dc.contributor.authorEun-Ji Lee-
dc.contributor.authorYeon Ju Kang-
dc.contributor.authorS Kim-
dc.contributor.authorS S Yoo-
dc.contributor.authorG M Lee-
dc.contributor.authorYeon-Gu Kim-
dc.date.accessioned2022-06-02T15:31:43Z-
dc.date.available2022-06-02T15:31:43Z-
dc.date.issued2022-
dc.identifier.issn0175-7598-
dc.identifier.urihttps://oak.kribb.re.kr/handle/201005/26103-
dc.description.abstractOptimizing appropriate signal peptides in mammalian cell-based protein production is crucial given that most recombinant proteins produced in mammalian cells are thought to be secreted proteins. Until now, most studies on signal peptide in mammalian cells have replaced native signal peptides with well-known heterologous signal peptides and bioinformatics-based signal peptides. In the present study, we successfully established an in vitro screening system for synthetic signal peptide in CHO cells by combining a degenerate codon-based oligonucleotides library, a site-specific integration system, and a FACS-based antibody detection assay. Three new signal peptides were screened using this new screening system, confirming to have structural properties as signal peptides by the SignalP web server, a neural network-based algorithm that quantifies the signal peptide-ness of amino acid sequences. The novel signal peptides selected in this study increased Fc-fusion protein production in CHO cells by increasing specific protein productivity, whereas they did not negatively affect cell growth. Particularly, the SP-#149 clone showed the highest qp, 0.73 ± 0.01 pg/cell/day from day 1 to day 4, representing a 1.47-fold increase over the native signal peptide in a serum-free suspension culture mode. In addition, replacing native signal peptide with the novel signal peptides did not significantly affect sialylated N-glycan formation, N-terminal cleavage pattern, and biological function of Fc-fusion protein produced in CHO cells. The overall results indicate the utility of a novel in vitro screening system for synthetic signal peptide for mammalian cell-based protein production. KEY POINTS: ? An in vitro screening system for synthetic signal peptide in mammalian cells was established ? This system combined a degenerate codon-based library, site-specific integration, and a FACS-based detection assay ? The novel signal peptides selected in this study could increase Fc-fusion protein production in mammalian cells.-
dc.publisherSpringer-
dc.titleDevelopment of an in vitro screening system for synthetic signal peptide in mammalian cell?based protein production-
dc.title.alternativeDevelopment of an in vitro screening system for synthetic signal peptide in mammalian cell?based protein production-
dc.typeArticle-
dc.citation.titleApplied Microbiology and Biotechnology-
dc.citation.number9-
dc.citation.endPage3582-
dc.citation.startPage3571-
dc.citation.volume106-
dc.contributor.affiliatedAuthorJong-Ho Park-
dc.contributor.affiliatedAuthorHoon Min Lee-
dc.contributor.affiliatedAuthorEun-Ju Jin-
dc.contributor.affiliatedAuthorEun-Ji Lee-
dc.contributor.affiliatedAuthorYeon Ju Kang-
dc.contributor.affiliatedAuthorYeon-Gu Kim-
dc.contributor.alternativeName박종호-
dc.contributor.alternativeName이훈민-
dc.contributor.alternativeName진은주-
dc.contributor.alternativeName이은지-
dc.contributor.alternativeName강연주-
dc.contributor.alternativeName김성균-
dc.contributor.alternativeName유성식-
dc.contributor.alternativeName이균민-
dc.contributor.alternativeName김연구-
dc.identifier.bibliographicCitationApplied Microbiology and Biotechnology, vol. 106, no. 9, pp. 3571-3582-
dc.identifier.doi10.1007/s00253-022-11955-6-
dc.subject.keywordSignal peptide-
dc.subject.keywordMammalian cells-
dc.subject.keywordScreening system-
dc.subject.keywordSite-specifc integration system-
dc.subject.keywordFc-fusion protein production-
dc.subject.localsignal peptide-
dc.subject.localSignal peptide-
dc.subject.localmammalian cells-
dc.subject.localMammalian cells-
dc.subject.localScreening system-
dc.subject.localscreening system-
dc.subject.localSite-specifc integration system-
dc.subject.localFc-fusion protein production-
dc.description.journalClassY-
Appears in Collections:
Division of Biomedical Research > Biotherapeutics Translational Research Center > 1. Journal Articles
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