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dc.contributor.authorZhang, F.
dc.contributor.authorWolff, M.W.
dc.contributor.authorWilliams, D.
dc.contributor.authorBusch, K.
dc.contributor.authorLang, S.C.
dc.contributor.authorMurhammer, D.W.
dc.contributor.authorLinhardt, Robert J.
dc.date2001
dc.date.accessioned2022-06-27T16:20:26Z
dc.date.available2022-06-27T16:20:26Z
dc.date.issued2001-01-01
dc.identifier.citationAffinity Purification of Secreted Alkaline Phosphatase Produced by the Baculovirus Expression Vector System, F. Zhang, M.W. Wolff, D. Williams, K. Busch, S.C. Lang, D.W. Murhammer, R.J. Linhardt, Applied Biochemistry and Biotechnology, 90, 125-136, 2001.
dc.identifier.issn2732289
dc.identifier.urihttps://doi.org/10.1385/ABAB:90:2:125
dc.identifier.urihttps://hdl.handle.net/20.500.13015/5855
dc.descriptionApplied Biochemistry and Biotechnology, 90, 125-136
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.abstractHuman secreted alkaline phosphatase (SEAP) was produced in a stably-transformed Spodoptera frugiperda Sf-9 insect cell line (Sfb4GalT) following infection with a recombinant Autographa californica multiple nuclear polyhedrovirus containing the SEAP gene under control of the polyhedrin promoter. An affinity chromatographic column prepared by linking 4-amino-benzylphosphonic acid to histidyl-expoxy-Sepharose was used to isolate SEAP from the cell supernatant following removal of cells and virus and 10-fold concentration through ultrafiltration. We found that the binding of SEAP on the affinity matrix follows the Langmuir isotherm model. In addition, either recycling SEAP sample through the column for 24 h or loading high SEAP concentrations resulted in a high-purity product. Some nonspecific binding of protein on the matrix occurred when low concentrations of SEAP sample were loaded. Finally, we found that SEAP binding occurs rapidly, i.e., within 30 min of adding the SEAP sample to the affinity matrix.
dc.description.sponsorshipNational Science Foundation
dc.description.urihttps://login.libproxy.rpi.edu/login?url=https://doi.org/10.1385/ABAB:90:2:125
dc.languageen_US
dc.language.isoENG
dc.relation.ispartofThe Linhardt Research Labs Online Collection
dc.relation.ispartofRensselaer Polytechnic Institute, Troy, NY
dc.relation.ispartofApplied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology
dc.relation.urihttps://harc.rpi.edu/
dc.subjectBiology
dc.subjectChemistry and chemical biology
dc.subjectChemical and biological engineering
dc.subjectBiomedical engineering
dc.titleAffinity Purification of Secreted Alkaline Phosphatase Produced by the Baculovirus Expression Vector System
dc.typeArticle
dcterms.accessRightshttps://login.libproxy.rpi.edu/login?url=https://doi.org/10.1385/ABAB:90:2:125
dcterms.isPartOfJournal
dcterms.isVersionOfhttps://doi.org/10.1385/ABAB:90:2:125
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.pages125-136
rpi.description.volume90


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