Study on the structure and function of glycans in target proteins for moving toward individualized medicine using antibody drugs

2016 Fiscal Year Final Research Report

• Project/Area Number: 26293037
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Medical pharmacy
• Research Institution: Yokohama City University (2015-2016); National Institute of Health Sciences (2014)
• Principal Investigator: Kawasaki Nana 横浜市立大学, 生命医科学研究科, 教授 (20186167)
• Co-Investigator(Kenkyū-buntansha): 小川 久美子 国立医薬品食品衛生研究所, 病理部, 部長 (70254282) ; 多田 稔 国立医薬品食品衛生研究所, 生物薬品部, 室長 (50506954) ; 石井 明子 国立医薬品食品衛生研究所, 生物薬品部, 室長 (50291117)
• Project Period (FY): 2014-04-01 – 2017-03-31
• Keywords: 糖鎖 / 質量分析 / EGF受容体 / 抗体医薬品

Outline of Final Research Achievements

For individualized medicine using monoclonal antibody drugs (mAbs)， it is important to clarify the diversity of target molecules for mAbs and the relationship between the diversity and efficacy and safety of mAb products. To elucidate the glycan diversity on target proteins for mAbs， we developed an analytical method for the site-specific glycosylation of membrane proteins; this method comprised the extraction of target proteins from the membrane fraction by immunoprecipitation with mAbs， trypsin digestion， glycopeptide enrichment by acetone precipitation， and LC/MS. Using the method， we successfully demonstrated the glycosylation at 5 sites of EGFR in A431 cells. It was suggested that Asn 528 is attached to unique glycans bearing both 7-9 GlcNAc and 2-6 Fuc residues. We also showed the site-specific glycosylation of the Fc gamma receptor IIIb expressed in Baby hamster kidney cells and in human serum.

Free Research Field

糖鎖生物学　質量分析学　バイオ医薬品