Improved preparation of group-specific component (Gc) protein to derive macrophage activating factor

Yuki Morita; Rong Wang; Xuyang Li; Tomonari Muramatsu; Masumi Ueda; Satoshi Hachimura; Sachiko Takahashi; Takuya Miyakawa; Masaru Tanokura

doi:10.1016/j.pep.2020.105714

Improved preparation of group-specific component (Gc) protein to derive macrophage activating factor

Protein Expr Purif. 2020 Nov:175:105714. doi: 10.1016/j.pep.2020.105714. Epub 2020 Jul 29.

Authors

Yuki Morita¹, Rong Wang², Xuyang Li³, Tomonari Muramatsu³, Masumi Ueda⁴, Satoshi Hachimura⁵, Sachiko Takahashi⁶, Takuya Miyakawa⁷, Masaru Tanokura⁸

Affiliations

¹ Laboratory of Basic Science on Healthy Longevity, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan; Medical Viara, 5-19 Akashi-cho, Chuo-ku, Tokyo, 104-0044, Japan; MAF Clinic, 5-19 Akashi-cho, Chuo-ku, Tokyo, 104-0044, Japan.
² Laboratory of Basic Science on Healthy Longevity, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan; Research Center for Food Safety, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
³ Laboratory of Basic Science on Healthy Longevity, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
⁴ Medical Viara, 5-19 Akashi-cho, Chuo-ku, Tokyo, 104-0044, Japan; MAF Clinic, 5-19 Akashi-cho, Chuo-ku, Tokyo, 104-0044, Japan.
⁵ Research Center for Food Safety, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan.
⁶ Medical Viara, 5-19 Akashi-cho, Chuo-ku, Tokyo, 104-0044, Japan; MAF Clinic, 5-19 Akashi-cho, Chuo-ku, Tokyo, 104-0044, Japan. Electronic address: info@mafclinic.com.
⁷ Laboratory of Basic Science on Healthy Longevity, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan. Electronic address: atmiya@mail.ecc.u-tokyo.ac.jp.
⁸ Laboratory of Basic Science on Healthy Longevity, Department of Applied Biological Chemistry, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo, 113-8657, Japan. Electronic address: amtanok@mail.ecc.u-tokyo.ac.jp.

PMID: 32738434
DOI: 10.1016/j.pep.2020.105714

Abstract

Cancer immunotherapy has recently attracted attention as an approach for cancer treatment through the activation of the immune system. Group-specific component (Gc) protein is a precursor for macrophage activating factor (GcMAF), which has a promising immunomodulatory effect on the suppression of tumor growth and angiogenesis. In this study, we successfully purified Gc protein from human serum using anion-exchange chromatography combined with affinity chromatography using a 25-OH-D₃-immobilized column. The purity of Gc protein reached 95.0% after anion-exchange chromatography. The known allelic variants of Gc protein are classified into three subtypes-Gc1F, Gc1S and Gc2. The fragment sequence of residues 412-424 determined according to their MS/MS spectra is available to evaluate the subtypes of Gc protein. The data showed that the Gc protein purified in this study consisted of the Gc1F and Gc2 subtypes. Our method improved the purity of Gc protein, which was not affected by the treatment to convert it into GcMAF using β-galactosidase- or neuraminidase-immobilized resin, and will be useful for biological studies and/or advanced clinical uses of GcMAF, such as cancer immunotherapy.

Keywords: Cancer immunotherapy; Gc globulin; Macrophage; Protein purification; Tumor.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Chromatography, Affinity*
Humans
Macrophage-Activating Factors* / chemistry
Macrophage-Activating Factors* / isolation & purification
Vitamin D-Binding Protein* / chemistry
Vitamin D-Binding Protein* / isolation & purification

Substances

GC protein, human
Macrophage-Activating Factors
Vitamin D-Binding Protein
vitamin D-binding protein-macrophage activating factor