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Highly efficient antibody purification with controlled orientation of protein A on magnetic nanoparticles

Abstract

In this study, we prepared the protein A grafted magnetic nanoparticles for industrial large-scale purification of antibodies with enhancement of binding capacity and immobilization by controlled orientation with chlorophenylsilane (CPTMS) on the surface. For site-specific immobilization of protein A, the genetically modified Protein A with cysteine residue was expressed in E.coli and purified by affinity chromatography. To improve surface area to volume and increase immobilization amount of protein A, chlorophenylsilane functionalized magnetic nanoparticles (CPTMS@MNPs) were prepared smaller nanoparticles with average diameter of 20 nm than commercial magnetic microparticles (Dynalbeads) with average size of 2.8 m. The CPTMS@MNPs showed the enhancement of Protein A immobilization and binding capacity to antibodies for 11.5-folds, and 7-fold higher than those of Dynalbeads, respectively. In addition, the CPTMS@MNPs retained about 80% of the initial protein binding capacity until the third stage of recycling. Therefore, protein A grafted CPTMS@MNPs may be useful to industrial large-scale purification of antibodies.

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Publication details

The article was received on 14 Sep 2017, accepted on 01 Nov 2017 and first published on 02 Nov 2017


Article type: Research Article
DOI: 10.1039/C7MD00468K
Citation: Med. Chem. Commun., 2017, Accepted Manuscript
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    Highly efficient antibody purification with controlled orientation of protein A on magnetic nanoparticles

    S. Kim, D. Sung and J. H. Chang, Med. Chem. Commun., 2017, Accepted Manuscript , DOI: 10.1039/C7MD00468K

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