| Enhanced production of human FcγRIIa receptor by high cell density cultivation of Escherichia coli. | |
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MedLine Citation:
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PMID: 21511037 Owner: NLM Status: Publisher |
Abstract/OtherAbstract:
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Human Fc receptors (FcγR) are membrane glycoproteins that are expressed on all immunologically active cells and have a well-defined role in regulating innate and adaptive immune responses by binding to the immunoglobulin G (IgG) antibody. Among the several classes of Fc receptors, FcγRIIa is the most widely expressed, and it serves as an important reagent in antibody engineering. Here, we report on high cell density cultivations (HCDC) of Escherichia coli for preparative scale production of FcγRIIa in a 6.6L bioreactor. Briefly, a pH-stat feeding strategy was employed, and two different cell densities (OD(600) of 46 and 100) were examined for the induction of FcγRIIa gene expression. When cells were induced at a high cell density (OD(600) of 100), the cell density increased to an OD(600) of 234 within 9h after induction, and a 2-fold higher production yield was obtained compared with that of induction at low cell density (OD(600) of 46). After simple purification steps including denaturation and refolding, 87.7mg of soluble FcγRIIa that was more than 95% pure was obtained from a 20-mL culture with high recovery yield (≈54%). The biological activity of purified FcγRIIa was also confirmed by evaluating its interaction with all subclasses of IgG antibodies using an ELISA bioassay. |
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Authors:
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Natarajan Velmurugan; Hee Sung Kim; Ki Jun Jeong |
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Publication Detail:
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Type: JOURNAL ARTICLE Date: 2011-4-12 |
Journal Detail:
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Title: Protein expression and purification Volume: - ISSN: 1096-0279 ISO Abbreviation: - Publication Date: 2011 Apr |
Date Detail:
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Created Date: 2011-4-22 Completed Date: - Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 9101496 Medline TA: Protein Expr Purif Country: - |
Other Details:
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Languages: ENG Pagination: - Citation Subset: - |
Copyright Information:
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Copyright © 2011. Published by Elsevier Inc. |
Affiliation:
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Department of Chemical and Biomolecular Engineering, KAIST, Yuseong-gu, Daejeon 305-701, Republic of Korea. |
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From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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