Purification of Fab and Fc using papain immobilized cryogel bioreactor separator system

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dc.authoridCorman, Mehmet Emin/0000-0001-9062-4201
dc.authoridUzun, Lokman/0000-0002-3971-7725
dc.contributor.authorArmutcu, Canan
dc.contributor.authorCorman, Mehmet Emin
dc.contributor.authorBayram, Engin
dc.contributor.authorUzun, Lokman
dc.date.accessioned2025-03-23T19:41:18Z
dc.date.available2025-03-23T19:41:18Z
dc.date.issued2020
dc.departmentSinop Üniversitesi
dc.description.abstractThe continuous bed bioreactor systems have been used for the production of protein therapeutics, such as IgG, using immobilized enzyme in biopharmaceutical applications. We developed macroporous poly(hydroxyethyl methacrylate-co-glycidyl methacrylate) cryogel-based bioreactor matrix using sodium dodecyl sulfate as surfactans in the presence of ethylene glycol dimethacrylate as cross linking agent by bulk polymerization. The developed polyGMA immobilized bioreactor with papain enzyme was used for specific fragmentation of immunoglobulin G. The catalysis efficiency for immobilized enzyme were investigated in comparison with free enzyme. The immobilized papain displayed broad catalytic activity over a variety of conditions, with maximal activity around pH 7.0 and 70 degrees C. The Michaelis-Menten kinetic constant (K-m), the maximum reaction velocity (V-max), and the catalytic efficiency (k(cat)) for free enzyme were 0.1097 mg/mL, 29.9 mg/mL/min, and 92.01 1/min, respectively, whereas for immobilized enzyme, K-m, V-max, and k(cat) values were 0.1078 mg/mL, 30.53 mg/mL/min, and 94.3 1/min, respectively. In a further step, after digestion, remarkable digestion products of bioreactor, Fab and Fc fragments, produced with immobilized papain bioreactors were analyzed in two ways by SDS-PAGE and reversed-phase HPLC; it was demonstrated that papain immobilized bioreactor successfully used for the digestion of human IgG with high activity. Therefore, the polyGMA cryogel immobilized with papain exhibited a very effective matrix for the bioreactor which can be considered as an alternative bioreactor matrix with great promise in biopharmaceutical applications.
dc.description.sponsorshipHacettepe University Scientific Research Project Coordination Unit [FHD-201816930]
dc.description.sponsorshipThis work was supported by the Hacettepe University Scientific Research Project Coordination Unit (Project number: FHD-201816930). L.U. would like to thank Turkish Academy of Sciences (TUBAGEBIP) for their invaluable support.
dc.identifier.doi10.1016/j.jchromb.2020.122396
dc.identifier.issn1570-0232
dc.identifier.issn1873-376X
dc.identifier.pmid33091678
dc.identifier.scopus2-s2.0-85092913684
dc.identifier.scopusqualityQ2
dc.identifier.urihttps://doi.org/10.1016/j.jchromb.2020.122396
dc.identifier.urihttps://hdl.handle.net/11486/6547
dc.identifier.volume1158
dc.identifier.wosWOS:000585264800002
dc.identifier.wosqualityQ2
dc.indekslendigikaynakWeb of Science
dc.indekslendigikaynakScopus
dc.indekslendigikaynakPubMed
dc.language.isoen
dc.publisherElsevier
dc.relation.ispartofJournal of Chromatography B-Analytical Technologies in the Biomedical and Life Sciences
dc.relation.publicationcategoryMakale - Uluslararası Hakemli Dergi - Kurum Öğretim Elemanı
dc.rightsinfo:eu-repo/semantics/closedAccess
dc.snmzKA_WOS_20250323
dc.subjectCryogel bioreactor
dc.subjectImmobilization
dc.subjectHydrophobic interaction
dc.subjectIgG
dc.subjectFab fragment
dc.subjectPapain
dc.titlePurification of Fab and Fc using papain immobilized cryogel bioreactor separator system
dc.typeArticle

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