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

作者：Liu, Jianguo^*; Chen, Changzhen; Ge, Baosheng; Lu, Jianren; Cui, Zhanfeng

来源：Journal of Membrane Science, 2013, 425: 98-104.

DOI：10.1016/j.memsci.2012.09.020

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

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.

• 出版日期2013-1-1
• 单位中国石油大学（华东）