Highly selective magnetic affinity purification of histidine-tagged proteins by Ni²⁺ carrying monodisperse composite microspheres

Abstract

A magnetic sorbent based on monodisperse-porous silica microspheres was developed for His-tagged protein purification by immobilized metal affinity chromatography.

Molecularly imprinted polymeric shell coated monodisperse‐porous silica microspheres as a stationary phase for microfluidic boronate affinity chromatography

“…By performing an elemental analysis, carbon and nitrogen contents of bare SiO 2 microspheres were determined as 0.11 and 0.46% w/w, respectively. Carbon and nitrogen contents of bare SiO 2 microspheres should come from the poly(MAA‐co‐EDMA) seed particles and TBAI used as the starting material and the stabilizer, respectively, in the synthesis . On the other hand, carbon and the nitrogen contents of MIPS@SiO 2 microspheres were found as 2.92 and 0.54% w/w, respectively.…”

“…Monodisperse‐porous silica microspheres (bare SiO 2 microspheres) were synthesized by a staged shape templated hydrolysis‐condensation method by using monodisperse‐porous poly(MAA‐co‐EDMA) microspheres as the seed particles .…”

Bifunctional nickel–iminodiacetic acid-core–shell silica nanoparticles for the exclusion of high molecular weight proteins and purification of His-tagged recombinant proteins

“…Currently, in most studies regarding protein structure and function, recombinant protein technology is widely used. 1 To produce these heterologous proteins different hosts such as Escherichia coli, Pichia pastoris, Saccharomyces cerevisiae, and insect or mammalian cells have been employed. 2 Biotechnology enables proteins to be expressed with a certain tag and histidine tags, mainly hexa-histidine (6His-tag), are the most used affinity tags for recombinant protein purication.…”

“…2 Biotechnology enables proteins to be expressed with a certain tag and histidine tags, mainly hexa-histidine (6His-tag), are the most used affinity tags for recombinant protein purication. 1,3 Immobilized metal ion affinity chromatography (IMAC) was rst developed by Porath in 1975 (ref. 4) and is one of the most effective procedures for purication of His-tagged proteins 5 and is based on the metal coordination interaction between the imidazole ring of histidine and divalent transition metal ions such as Ni 2+ , Cu 2+ , Zn 2+ or Co 2+ .…”

“…In this context, silica nanoparticles have appealing physical and chemical properties as a sorbent, due to its large surface area, low toxicity, high stability and particularly because it can be chemically modied with several compounds. 1,[16][17][18][19] The aim of this work was to synthetize functionalized core-shell silica nanoparticles with IDA and a polyacrylamide shell, thus only low molecular weight (LMW) His-tagged proteins that pass through the polymeric shell to interact with the immobilized metal ions, while other HMW host proteins with inherent histidine-rich region, are excluded.…”