Preparation of nitrilotriacetic acid/Co2+-linked, silica/boron-coated magnetite nanoparticles for purification of 6 x histidine-tagged proteins.
In this report, we describe the preparation of novel nitrilotriacetic acid/Co2+-linked, silica/boron-coated magnetite nanoparticles for purification of 6 x His-tagged proteins. The nanoparticles were approximately 200 nm in size and were stable against hydrochloric acid and had negligible non-specific binding for protein. Elimination of non-specific binding by nucleic acids was readily achieved by digestion of samples with DNase and RNase. The modified nanoparticles were used to purify two model proteins: one had a C-terminal 6 x His tag, and the other had an internal 6 x His tag. Both proteins were purified within one hour into single band purity on sodium dodecyl sulfate-polyacrylamide electrophoresis gel.
Liao
Yiqun
Y
Molecular Diagnostics Laboratory, Department of Biomedical Sciences, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, China.
Cheng
Yangjian
Y
Li
Qingge
Q
D003035:Cobalt / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D052203:Ferrosoferric Oxide / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D053768:Metal Nanoparticles; D046529:Microscopy, Electron, Transmission; D009571:Nitrilotriacetic Acid / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012822:Silicon Dioxide / Q000737:chemistry*
D011506:Proteins; D003035:Cobalt; D006639:Histidine; D012822:Silicon Dioxide; D009571:Nitrilotriacetic Acid; D052203:Ferrosoferric Oxide