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Carboxymethylated polyethylenimine modified magnetic nanoparticles specifically for purification of His‐tagged protein
Author(s) -
Chang Mengmeng,
Qin Qian,
Wang Bohong,
Xia Tian,
lv Wangjie,
Sun Xiaoshan,
Shi Xianzhe,
Xu Guowang
Publication year - 2019
Publication title -
journal of separation science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.72
H-Index - 102
eISSN - 1615-9314
pISSN - 1615-9306
DOI - 10.1002/jssc.201800969
Subject(s) - polyethylenimine , magnetic nanoparticles , nanoparticle , adsorption , materials science , chemistry , chromatography , nanotechnology , chemical engineering , organic chemistry , biochemistry , transfection , engineering , gene
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‐Ni 2+ @SiO 2 @Fe 3 O 4 (CMPEI=carboxymethylatedpolyethyleneimine) magnetic nanoparticles composed of a strong magnetic core of Fe 3 O 4 and a Ni 2+ ‐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 SiO 2 @Fe 3 O 4 . The resulting CMPEI‐Ni 2+ @SiO 2 @Fe 3 O 4 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‐Ni 2+ @SiO 2 @Fe 3 O 4 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‐Ni 2+ @SiO 2 @Fe 3 O 4 magnetic nanoparticles have great potential for application in biotechnological fields.