A novel membrane based process to isolate recombinant human chemokine receptor CCR3 produced in Escherichia coli

Jianguo Liu, Changzhen Chen, Baosheng Ge, Jianren Lu, Zhanfeng Cui

    Research output: Contribution to journalArticlepeer-review

    Abstract

    The isolation of recombinant human chemokine receptor CCR3, which fused to maltose binding protein (MBP), has been conducted using ultrafiltration with 300. kDa molecular weight cut-off polyethersulfone membranes. The effects of ultrafiltration operating conditions on MBP-CCR3 stability and transmission were quantified using dot blot analysis and parameter scanning ultrafiltration respectively. These conditions included solution pH, ionic strength, stirring speed, permeate flux, and detergent (Fos choline-14) concentration. Under optimized conditions, the MBP-CCR3 purity obtained in the retentate was about 96% and the recovery of MBP-CCR3 was close to 89% after ultrafiltration. The resulting MBP-CCR3 product was then analyzed by isoelectric focusing, sodium dodecyl sulfate-polyacrylamide gel eletrophoresis and circular dichroism, to confirm its isoelectric point, molecular weight and molecular secondary structure. To our knowledge, this is the first paper of the isolation of a G protein coupled receptor (GPCR) using ultrafiltration alone. © 2012 Elsevier B.V.
    Original languageEnglish
    Pages (from-to)98-104
    Number of pages6
    JournalJournal of Membrane Science
    Volume425-426
    DOIs
    Publication statusPublished - 1 Jan 2013

    Keywords

    • CC chemokine receptor 3
    • Membrane protein
    • Purification
    • Ultrafiltration

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