Thermostabilization of Fab and its application to medicine

2019 Fiscal Year Final Research Report

• Project/Area Number: 16K08922
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Applied pharmacology
• Research Institution: Sojo University
• Principal Investigator: Ohkuri Takatoshi 崇城大学, 薬学部, 教授 (70346807)
• Co-Investigator(Kenkyū-buntansha): 安楽 誠 崇城大学, 薬学部, 教授 (60398245) ; 中村 仁美 崇城大学, 薬学部, 助教 (60510691)
• Project Period (FY): 2016-04-01 – 2020-03-31
• Keywords: 抗体工学 / Fab / 抗体医薬 / 糖鎖エンジニアリング / ジスルフィド結合 / 安定化

Outline of Final Research Achievements

In recent years, antibody drugs have been actively developed and are highly effective against many diseases. In this study, we produced a recombinant protein of Fab molecule of human monoclonal antibody drug adalimumab and produced a modified Fab whose functionality was improved by introducing an amino acid mutation. As a result of designing a novel intermolecular SS bond introduction mutation for the purpose of improving stability, 12 mutants were successfully constructed. we have successfully demonstrated that a Fab mutant with a novel interchain SS bond (H:V177C-L:Q160C) and one free cysteine at the C-terminal end can be PEGylated without changes in functionality. N-glycosylation site was introduced at H-chain constant region of adalimumab Fab through site-directed mutagenesis. We demonstrated that glycosylated Fab can prevent protein aggregation.

Free Research Field

タンパク質工学

Academic Significance and Societal Importance of the Research Achievements

抗体医薬品の欠点の１つは生産コストが高いという点である。本研究において抗体医薬品アダリムアブのFabについて機能を高めた改変Fabを酵母を用いて作製することに成功した。酵母によって安価に生産できるFabが抗体医薬として応用できれば薬価を抑えることが期待できる。また高機能化によりIgG分子より優れた効果を生み出す可能性もうかがえた。本研究成果は、Fabの抗体医薬品への応用につながる重要な知見をもたらしたであろう。