title: Facile and high-efficient immobilization of histidine-tagged multimeric protein G on magnetic nanoparticles.
abstract: This work reports the high-efficient and one-step immobilization of multimeric protein G on magnetic nanoparticles. The histidine-tagged (His-tag) recombinant multimeric protein G was overexpressed in Escherichia coli BL21 by the repeated linking of protein G monomers with a flexible linker. High-efficient immobilization on magnetic nanoparticles was demonstrated by two different preparation methods through the amino-silane and chloro-silane functionalization on silica-coated magnetic nanoparticles. Three kinds of multimeric protein G such as His-tag monomer, dimer, and trimer were tested for immobilization efficiency. For these tests, bicinchoninic acid (BCA) assay was employed to determine the amount of immobilized His-tag multimeric protein G. The result showed that the immobilization efficiency of the His-tag multimeric protein G of the monomer, dimer, and trimer was increased with the use of chloro-silane-functionalized magnetic nanoparticles in the range of 98% to 99%, rather than the use of amino-silane-functionalized magnetic nanoparticles in the range of 55% to 77%, respectively.
authors.affiliation: Korea Institute of Ceramic Engineering and Technology, Seoul 153-801, South Korea.
authors.forename: Jiho
authors.initials: J
authors.lastname: Lee
authors.affiliation: Korea Institute of Ceramic Engineering and Technology, Seoul 153-801, South Korea.
authors.forename: Jeong Ho
authors.initials: JH
authors.lastname: Chang
keywords: Efficiency; Histidine-tag; Immobilization; Magnetic nanoparticle; Protein G