Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/81450
Full metadata record
DC FieldValueLanguage
dc.contributor.authorChng, Jakeen
dc.contributor.authorWang, Tianhuaen
dc.contributor.authorNian, Ruien
dc.contributor.authorLau, Allyen
dc.contributor.authorHoi, Kong Mengen
dc.contributor.authorYang, Yuanshengen
dc.contributor.authorHo, Steven C. L.en
dc.contributor.authorGagnon, Peteren
dc.contributor.authorBi, Xuezhien
dc.date.accessioned2016-06-23T08:52:59Zen
dc.date.accessioned2019-12-06T14:31:15Z-
dc.date.available2016-06-23T08:52:59Zen
dc.date.available2019-12-06T14:31:15Z-
dc.date.issued2015en
dc.identifier.citationChng, J., Wang, T., Nian, R., Lau, A., Hoi, K. M., Ho, S. C. L., et al. (2015). Cleavage efficient 2A peptides for high level monoclonal antibody expression in CHO cells. mAbs, 7(2), 403-412.en
dc.identifier.issn1942-0862en
dc.identifier.urihttps://hdl.handle.net/10356/81450-
dc.description.abstractLinking the heavy chain (HC) and light chain (LC) genes required for monoclonal antibodies (mAb) production on a single cassette using 2A peptides allows control of LC and HC ratio and reduces non-expressing cells. Four 2A peptides derived from the foot-and-mouth disease virus (F2A), equine rhinitis A virus (E2A), porcine teschovirus-1 (P2A) and Thosea asigna virus (T2A), respectively, were compared for expression of 3 biosimilar IgG1 mAbs in Chinese hamster ovary (CHO) cell lines. HC and LC were linked by different 2A peptides both in the absence and presence of GSG linkers. Insertion of a furin recognition site upstream of 2A allowed removal of 2A residues that would otherwise be attached to the HC. Different 2A peptides exhibited different cleavage efficiencies that correlated to the mAb expression level. The relative cleavage efficiency of each 2A peptide remains similar for expression of different IgG1 mAbs in different CHO cells. While complete cleavage was not observed for any of the 2A peptides, GSG linkers did enhance the cleavage efficiency and thus the mAb expression level. T2A with the GSG linker (GT2A) exhibited the highest cleavage efficiency and mAb expression level. Stably amplified CHO DG44 pools generated using GT2A had titers 357, 416 and 600 mg/L for the 3 mAbs in shake flask batch cultures. Incomplete cleavage likely resulted in incorrectly processed mAb species and aggregates, which were removed with a chromatin-directed clarification method and protein A purification. The vector and methods presented provide an easy process beneficial for both mAb development and manufacturing.en
dc.description.sponsorshipASTAR (Agency for Sci., Tech. and Research, S’pore)en
dc.format.extent34 p.en
dc.language.isoenen
dc.relation.ispartofseriesmAbsen
dc.rights© 2014 Landes Bioscience. This is the author created version of a work that has been peer reviewed and accepted for publication in mAbs, published by Taylor & Francis on behalf of Landes Bioscience. It incorporates referee’s comments but changes resulting from the publishing process, such as copyediting, structural formatting, may not be reflected in this document. The published version is available at: [http://dx.doi.org/10.1080/19420862.2015.1008351].en
dc.subjectmonoclonal antibodyen
dc.subject2A peptideen
dc.subjectfurinen
dc.subjectCHOen
dc.subjectcleavage efficiencyen
dc.subjectGSG linkeren
dc.titleCleavage efficient 2A peptides for high level monoclonal antibody expression in CHO cellsen
dc.typeJournal Articleen
dc.contributor.schoolSchool of Chemical and Biomedical Engineeringen
dc.identifier.doi10.1080/19420862.2015.1008351en
dc.description.versionAccepted versionen
dc.identifier.pmid25621616-
item.grantfulltextopen-
item.fulltextWith Fulltext-
Appears in Collections:SCBE Journal Articles
Files in This Item:
File Description SizeFormat 
Cleavage efficient 2A peptides for high level monoclonal antibody expression in CHO cells.pdf845.68 kBAdobe PDFThumbnail
View/Open

SCOPUSTM   
Citations 5

83
Updated on Feb 2, 2023

Web of ScienceTM
Citations 5

79
Updated on Feb 4, 2023

Page view(s) 50

425
Updated on Feb 5, 2023

Download(s) 10

373
Updated on Feb 5, 2023

Google ScholarTM

Check

Altmetric


Plumx

Items in DR-NTU are protected by copyright, with all rights reserved, unless otherwise indicated.