id,collection,dc.contributor.author[],dc.date.accessioned[],dc.date.available[],dc.date.issued[],dc.description.abstract[en],dc.format.extent[],dc.identifier.uri,dc.identifier.uri[],dc.language[],dc.rights.uri[],dc.subject.ddc[],dc.subject[],dc.title[],dc.type[],dcterms.accessRights.openaire,dcterms.bibliographicCitation.doi[],dcterms.bibliographicCitation.url[],dcterms.bibliographicCitation[],refubium.affiliation.other[],refubium.affiliation[de],refubium.mycore.derivateId[],refubium.mycore.fudocsId[],refubium.note.author[],refubium.resourceType.isindependentpub[] "63d3e8df-1e3e-4ff9-bebc-fe0935a1be52","fub188/16","Montacir, Othman||Montacir, Houda||Eravci, Murat||Springer, Andreas||Hinderlich, Stephan||Mahboudi, Fereidoun||Saadati, Amirhossein||Parr, Maria Kristina","2018-06-08T11:07:17Z","2018-03-22T13:55:33.225Z","2018","Eptacog alfa (NovoSeven®) is a vitamin K-dependent recombinant Factor VIIa produced by genetic engineering from baby hamster kidney (BHK) cells as a single peptide chain of 406 residues. After activation, it consists of a light chain (LC) of 152 amino and a heavy chain (HC) of 254 amino acids. Recombinant FVIIa undergoes many post-translational modifications (PTMs). The first ten glutamic acids of the N-terminal moiety are γ-carboxylated, Asn145 and Asn322 are N-glycosylated, and Ser52 and Ser60 are O-glycosylated. A head-to-head biosimilarity study was conducted for the originator and the first biosimilar AryoSeven™ to evaluate comparable bioengineering. Physicochemical properties were analyzed based on mass spectrometry, including intact mass, PTMs and higher-order structure. Both biotherapeutics exhibit a batch-to-batch variability in their N-glycan profiles. N-Glycopeptide analysis with UHPLC- QTOF-MSE confirmed N-glycosylation sites as well as two different O-glycopeptide sites. Ser60 was found to be O-fucosylated and Ser52 had O-glucose or O-glucose-(xylose)1,2 motifs as glycan variants. Ion mobility spectrometry (TWIMS) and NMR spectroscopy data affirm close similarity of the higher-order structure of both biologicals. Potency of the biodrugs was analyzed by a coagulation assay demonstrating comparable bioactivity. Consequently, careful process optimization led to a stable production process of the biopharmaceuticals.","16 Seiten","http://dx.doi.org/10.17169/refubium-24941","https://refubium.fu-berlin.de/handle/fub188/21653","eng","http://creativecommons.org/licenses/by/4.0/","600 Technik, Medizin, angewandte Wissenschaften::610 Medizin und Gesundheit::615 Pharmakologie, Therapeutik||500 Naturwissenschaften und Mathematik::570 Biowissenschaften; Biologie::572 Biochemie","biopharmaceutical||biosimilar||mass spectrometry||physicochemical characterization||coagulation assay","Bioengineering of rFVIIa Biopharmaceutical and Structure Characterization for Biosimilarity Assessment","Wissenschaftlicher Artikel","open access","10.3390/bioengineering5010007","http://doi.org/10.3390/bioengineering5010007","Bioengineering 5 (2018), 7","Institut für Chemie und Biochemie:::bdaf1c47-f341-464f-b012-0d24067d34b8:::600","Biologie, Chemie, Pharmazie","FUDOCS_derivate_000000009573","FUDOCS_document_000000029385","Der Artikel wurde in einer Open-Access-Zeitschrift publiziert.","no"