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) fibers was synthesized by a simple method based on the coordination effect between Ni and carboxyl group. The nickel content of the CCC-Ni fibers was determined to be 5 times larger than that of Ni-immobilized sulfhydryl cotton fiber (SCF-Ni) fibers developed in our previous work. The prepared CCC-Ni 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 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 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.