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dc.contributor.authorWolff, Michael W.
dc.contributor.authorZhang, Fuming
dc.contributor.authorRoberg, Jeff J.
dc.contributor.authorCaldwell, Elizabeth E.O.
dc.contributor.authorKaul, Patrick R.
dc.contributor.authorSerrahn, Jill N.
dc.contributor.authorMurhammer, David W.
dc.contributor.authorLinhardt, Robert J.
dc.contributor.authorWeiler, John M.
dc.date2001
dc.date.accessioned2022-06-27T16:19:41Z
dc.date.available2022-06-27T16:19:41Z
dc.date.issued2001-01-01
dc.identifier.citationExpression of C1 Esterase Inhibitor by the Baculovirus Expression System: Preparation, Purification and Characterization, M.W. Wolff, F. Zhang, J.J. Roberg, E.E.O. Caldwell, P.R. Kaul, J.N. Serrahn, D.W. Murhammer, R.J. Linhardt, J.M. Weiler, Protein Expression and Purification, 22 414-421, 2001.
dc.identifier.issn10465928
dc.identifier.urihttps://doi.org/10.1006/prep.2001.1461
dc.identifier.urihttps://hdl.handle.net/20.500.13015/5853
dc.descriptionProtein Expression and Purification, 22 414-421
dc.descriptionNote : if this item contains full text it may be a preprint, author manuscript, or a Gold OA copy that permits redistribution with a license such as CC BY. The final version is available through the publisher’s platform.
dc.description.abstractC1 esterase inhibitor (C1INH) is an important regulator of the classical complement pathway. Hereditary deficiency of C1INH causes angioedema of the skin, gut, and respiratory tissues that may be fatal. C1INH replacement therapy may be lifesaving for patients with this disorder. The objective of this study was to evaluate the use of the baculovirus expression vector system for mass producing biologically active human recombinant (rC1INH). A recombinant baculovirus was constructed coding the human native (nC1INH) sequence under control of the polyhedrin promoter. Spodoptera frugiperda Sf-9 insect cells were infected with this recombinant baculovirus in a medium-scale (10-L) bioreactor to produce rC1INH with a specific activity of 45 U/mg. Purification of rC1INH from the culture harvested at 60 h postinfection yielded 5.9 microg rC1INH/mL supernatant of a 75-kDa product with a specific activity of 31,000 U/mg purified rC1INH compared to 71,000 U/mg purified nC1INH from human serum using the same procedure. This rC1INH was about 25 kDa smaller than nC1INH, suggesting that Sf-9 cells express underglycosylated rC1INH. Glycan analysis showed that both N-glycan and O-glycan chains were present in rC1INH. The N-glycan chains, released using PNGaseF and fluorescently labeled, were analyzed using exoglycosidase treatment and capillary electrophoresis. Their high-mannose structure was consistent with the known failure of the insect cell glycosylation pathway to afford the fully elaborated biantennary structures found on human native nC1INH.
dc.description.sponsorshipNational Science Foundation
dc.description.urihttps://login.libproxy.rpi.edu/login?url=https://doi.org/10.1006/prep.2001.1461
dc.languageen_US
dc.language.isoENG
dc.relation.ispartofThe Linhardt Research Labs Online Collection
dc.relation.ispartofRensselaer Polytechnic Institute, Troy, NY
dc.relation.ispartofProtein Expression and Purification
dc.relation.urihttps://harc.rpi.edu/
dc.subjectBiology
dc.subjectChemistry and chemical biology
dc.subjectChemical and biological engineering
dc.subjectBiomedical engineering
dc.titleExpression of C1 Esterase Inhibitor by the Baculovirus Expression System: Preparation, Purification and Characterization
dc.typeArticle
dcterms.accessRightshttps://login.libproxy.rpi.edu/login?url=https://doi.org/10.1006/prep.2001.1461
dcterms.isPartOfJournal
dcterms.isVersionOfhttps://doi.org/10.1006/prep.2001.1461
dc.rights.holderIn Copyright : this Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). https://rightsstatements.org/page/InC/1.0/
dc.creator.identifierhttps://orcid.org/0000-0003-2219-5833
dc.relation.departmentThe Linhardt Research Labs.
dc.relation.departmentThe Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
rpi.description.pages414-421
rpi.description.volume22


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