Carboxymethylated polyethylenimine modified magnetic nanoparticles specifically for purification of His-tagged protein.
Mengmeng ChangQian QinBohong WangTian XiaWangjie LvXiaoshan SunXianzhe ShiGuowang XuPublished in: Journal of separation science (2018)
Employing immobilized metal-ion affinity chromatography and magnetic separation could ideally provide a useful analytical strategy for purifying His-tagged protein. In the current study, a facile route was designed to prepare CMPEI-Ni2+ @SiO2 @Fe3 O4 (CMPEI=carboxymethylated polyethyleneimine) magnetic nanoparticles composed of a strong magnetic core of Fe3 O4 and a Ni2+ -immobilized carboxymethylated polyethyleneimine coated outside shell, which was formed by electrostatic interactions between polyanionic electrolyte of carboxymethylated polyethyleneimine and positively charged surface of 3-(trimethoxysilyl)propylamin modified SiO2 @Fe3 O4 . The resulting CMPEI-Ni2+ @SiO2 @Fe3 O4 composite nanoparticles displayed well-uniform structure and high magnetic responsiveness. Hexa His-tagged peptides and purified His-tagged recombinant retinoid X receptor alpha were chosen as the model samples to evaluate the adsorption, capacity, and reusability of the composite nanoparticles. The results demonstrated the CMPEI-Ni2+ @SiO2 @Fe3 O4 nanoparticles possessed rapid adsorption, large capacity, and good recyclability. The obtained nanoparticles were further used to purify His-tagged protein in practical environment. It was found that the nanoparticles could selectively capture His-tagged recombinant retinoid X receptor protein from complex cell lysate. Owing to its easy synthesis, large binding capacity, and good reusability, the prepared CMPEI-Ni2+ @SiO2 @Fe3 O4 magnetic nanoparticles have great potential for application in biotechnological fields.
Keyphrases
- magnetic nanoparticles
- binding protein
- amino acid
- protein protein
- metal organic framework
- molecularly imprinted
- liquid chromatography
- mass spectrometry
- transition metal
- single cell
- ionic liquid
- risk assessment
- walled carbon nanotubes
- bone marrow
- high speed
- gold nanoparticles
- highly efficient
- aqueous solution
- high resolution
- ms ms