Facial synthesis of nickel(II)-immobilized carboxyl cotton chelator for purification of histidine-tagged proteins
Immobilized metal affinity chromatography (IMAC) technique is frequently used in the purification of histidine-tagged (His-tagged) recombinant proteins. In this study, nickel(II)-immobilized carboxyl cotton chelator (CCC-Ni 2+ ) fibers was synthesized by a simple method based on the coordination effect between Ni 2+ and carboxyl group. The nickel content of the CCC-Ni 2+ fibers was determined to be 5 times larger than that of Ni 2+ -immobilized sulfhydryl cotton fiber (SCF-Ni 2+ ) fibers developed in our previous work. The prepared CCC-Ni 2+ fibers were then applied for the selective and rapid separation of His-tagged protein from escherichia coli ( E. coli ) cell lysates on the basis of the high affinity of Ni 2+ to 6 × His with a lab-in-syringe format. Benefiting from the good biological compatibility and high nickel content, the results showed that CCC-Ni 2+ fibers were able to selectively capture His-tagged proteins from complex E. coli cell lysates and exhibited a relatively large adsorption capacity toward His-tagged protein. The recoveries of His-tagged GFP in E. coli cell lysates were in the range of 89.8%-106.7% with the relative standard deviations (RSDs) less than 9.4% (intra-day) and 10.3% (inter-day). Taken together, this efficient approach for the purification of recombinant proteins extends the application of CCC-based fibrous materials in biological analysis.
危险品化学品经营许可证（带存储）