Development of highly sensitive imaging method by use of functional single-chain antibodies that discriminate the structure of advanced glycation endproduct

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K15321
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Applied pharmacology
• Research Institution: Kumamoto University
• Principal Investigator: MORIOKA HIROSHI 熊本大学, 大学院生命科学研究部(薬), 教授 (20230097)
• Co-Investigator(Kenkyū-buntansha): 小橋川 敬博 熊本大学, 大学院生命科学研究部(薬), 准教授 (90455600) ; 佐藤 卓史 熊本大学, 大学院生命科学研究部(薬), 助教 (70555755)
• Co-Investigator(Renkei-kenkyūsha): MAENAKA Katsumi 北海道大学, 大学院薬学研究院, 教授 (10322752) ; MARUYAMA Toru 熊本大学, 大学院生命科学研究部, 教授 (90423657)
• Research Collaborator: FUKUDA Natsuki 熊本大学, 薬学教育部, 大学院生
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: 機能性一本鎖抗体 / 終末糖化産物 / 蛍光プローブ / 時間分解蛍光測定法 / クリックケミストリー

Outline of Final Research Achievements

We have produced the single-chain antibody (scFv) fused protein that recognizes a kind of advanced glycation end-product (AGE), GA-pyridine, and conducted research on development of in vivo imaging method for AGE localization in the body. The mutant scFv clones that exhibited higher affinity for antigen and thermal stability have been obtained using a combination of a phage display system and random mutagenesis. In order to improve stability and retentivity of the scFvs in blood, the scFv/human serum albumin (HSA) fused protein have been prepared by using transpeptidase Sortase A and consequently the improvement was confirmed in animal experiments. We have prepared scFv containing lanthanoid binding peptide (LBP) and tried to introduce lanthanoid ion usable for time-resolution fluorescent measurement in LBP, however could not observe fluorescence. Currently, we are carrying out introduction of fluorescent substance by click chemistry.

Free Research Field

タンパク質工学