| High-gradient magnetic affinity separation of trypsin from porcine pancreatin. | |
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MedLine Citation:
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PMID: 12115419 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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We introduce a robust and scale-flexible approach to macromolecule purification employing tailor-made magnetic adsorbents and high-gradient magnetic separation technology adapted from the mineral processing industries. Detailed procedures for the synthesis of large quantities of low-cost defined submicron-sized magnetic supports are presented. These support materials exhibit unique features, which facilitate their large-scale processing using high magnetic field gradients, namely sufficiently high magnetization, a relatively narrow particle size distribution and ideal superparamagnetism. Following systematic optimization with respect to activation chemistry, spacer length and ligand density, conditions for preparation of effective high capacity (Q(max) = 120 mg g(-1)) strongly interacting (Kd < 0.3 microm) trypsin-binding adsorbents based on immobilized benzamidine were established. In small-scale studies approximately 95% of the endogenous trypsin present in a crude porcine pancreatin feedstock was recovered with a purification factor of approximately 4.1 at the expense of only a 4% loss in alpha-amylase activity. Efficient recovery of trypsin from the same feedstock was demonstrated at a vastly increased scale using a high-gradient magnetic separation system to capture loaded benzamidine-linked adsorbents following batch adsorption. With the aid of a simple recycle loop over 80% of the initially adsorbed trypsin was recovered in-line with an overall purification factor of approximately 3.5. |
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Authors:
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Jürgen J Hubbuch; Owen R T Thomas |
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Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't |
Journal Detail:
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Title: Biotechnology and bioengineering Volume: 79 ISSN: 0006-3592 ISO Abbreviation: Biotechnol. Bioeng. Publication Date: 2002 Aug |
Date Detail:
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Created Date: 2002-07-12 Completed Date: 2003-01-17 Revised Date: 2006-11-15 |
Medline Journal Info:
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Nlm Unique ID: 7502021 Medline TA: Biotechnol Bioeng Country: United States |
Other Details:
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Languages: eng Pagination: 301-13 Citation Subset: IM |
Copyright Information:
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Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 79: 301-313, 2002. |
Affiliation:
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Center for Process Biotechnology, BioCentrum-DTU, Technical University of Denmark, Building 223, DK-2800, Kgs. Lyngby, Denmark. |
Export Citation:
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APA/MLA Format Download EndNote Download BibTex |
| MeSH Terms | |
Descriptor/Qualifier:
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Adsorption Animals Benzamidines / chemistry* Cattle Macromolecular Substances Magnetics* Models, Chemical Pancreatin / chemistry* Pilot Projects Quality Control Sensitivity and Specificity Swine Trypsin / isolation & purification* |
| Chemical | |
Reg. No./Substance:
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0/Benzamidines; 0/Macromolecular Substances; 618-39-3/benzamidine; 8049-47-6/Pancreatin; EC 3.4.21.4/Trypsin |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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