Characterization of metal affinity of green fluorescent protein and its purification through salt promoted, immobilized metal affinity chromatography

by Li, Y.; Agrawal, A.; Sakon, J.; Beitle, R. R.

Immobilized metal affinity chromatography (IMAC) was investigated as a method of recovery for green fluorescent protein (GFPuv). It was found that in the absence of genetic modification to enhance metal affinity, GFPuv displayed strong metal affinity to Cu(II) and Ni(II), and weak or negligible affinity to Zn(II) and Co(II). Changes in the mobile phase NaCl concentration during Ni(II)-IMAC strongly affected purity and yield of GFPuv, with fine resolution under higher NaCl concentrations. Finally, IMAC via Cu(II) and Zn(II) with intervening diafiltration was used to recover GFPuv with high yield and purity.

Journal
Journal of Chromatography A
Volume
909
Issue
2
Year
2001
Start Page
183-190
URL
https://dx.doi.org/10.1016/s0021-9673(00)01100-6
ISBN/ISSN
1873-3778; 0021-9673
DOI
10.1016/s0021-9673(00)01100-6