TY - JOUR
T1 - Immobilized-metal affinity chromatography adsorbent with paramagnetism and its application in purification of histidine-tagged proteins
AU - Feng, Guodong
AU - Hu, Daodao
AU - Yang, Lei
AU - Cui, Yali
AU - Cui, Xin Ai
AU - Li, Hong
PY - 2010/8/17
Y1 - 2010/8/17
N2 - A new method for synthesis of an Immobilized-Metal Affinity chromatography (IMAC) adsorbent with superparamagnetism (Fe3O4/SiO 2-GPTMS-Asp-Co) was reported in this paper. Fe3O 4 nanoparticles were first modified by SiO2 to form the core-shell Fe3O4/SiO2 with superparamagnetism, the core-shell microspheres were then successively treated by 3-glycidoxypropyltrimethoxysilane (GPTMS), l-aspartic acid (l-Asp) and 2-bromoacetic acid to form Fe3O4/SiO2-GPTMS-Asp nanoparticles with tetradentate ligands. Finally, the IMAC adsorbent with superparamagnetism, Fe3O4/SiO2-GPTMS-Asp-Co, was finally obtained by the coordination of Co2+ with the resulting nanoparticles. The intermediates and product obtained from the process mentioned above were characterized by TEM, SEM, XRD, XPS, FT-IR, TGA, AAS, EDS, element analysis and magnetic hysteresis loop. Fe3O4/SiO 2-GPTMS-Asp-Co nanoparticles used as IMAC adsorbent to separate the recombinant 6-histidine-tagged gp41 protein were investigated. The result indicates that the IMAC adsorbent we prepared has outstanding advantages in the separation of the 6-histidine-tagged proteins from the crude bacterial lysate, such as simple operation, high selectivity and capacity.
AB - A new method for synthesis of an Immobilized-Metal Affinity chromatography (IMAC) adsorbent with superparamagnetism (Fe3O4/SiO 2-GPTMS-Asp-Co) was reported in this paper. Fe3O 4 nanoparticles were first modified by SiO2 to form the core-shell Fe3O4/SiO2 with superparamagnetism, the core-shell microspheres were then successively treated by 3-glycidoxypropyltrimethoxysilane (GPTMS), l-aspartic acid (l-Asp) and 2-bromoacetic acid to form Fe3O4/SiO2-GPTMS-Asp nanoparticles with tetradentate ligands. Finally, the IMAC adsorbent with superparamagnetism, Fe3O4/SiO2-GPTMS-Asp-Co, was finally obtained by the coordination of Co2+ with the resulting nanoparticles. The intermediates and product obtained from the process mentioned above were characterized by TEM, SEM, XRD, XPS, FT-IR, TGA, AAS, EDS, element analysis and magnetic hysteresis loop. Fe3O4/SiO 2-GPTMS-Asp-Co nanoparticles used as IMAC adsorbent to separate the recombinant 6-histidine-tagged gp41 protein were investigated. The result indicates that the IMAC adsorbent we prepared has outstanding advantages in the separation of the 6-histidine-tagged proteins from the crude bacterial lysate, such as simple operation, high selectivity and capacity.
KW - Immobilized-meta affinity chromatography (IMAC)
KW - Magnetic materials
KW - Purification
KW - Sol-gel growth
KW - Tetradentate ligand
UR - https://www.scopus.com/pages/publications/77955514350
U2 - 10.1016/j.seppur.2010.06.013
DO - 10.1016/j.seppur.2010.06.013
M3 - 文章
AN - SCOPUS:77955514350
SN - 1383-5866
VL - 74
SP - 253
EP - 260
JO - Separation and Purification Technology
JF - Separation and Purification Technology
IS - 2
ER -