Development of Tag-assisted Phage Display for screening of peptide drug

2020 Fiscal Year Final Research Report

• Project/Area Number: 19K15696
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 37010:Bio-related chemistry
• Research Institution: Tokyo Institute of Technology
• Principal Investigator: MIKI Takayuki 東京工業大学, 生命理工学院, 助教 (20823991)
• Project Period (FY): 2019-04-01 – 2021-03-31
• Keywords: ファージディスプレイ / 膜蛋白質 / His-tag / Ni(II)錯体 / ペプチド

Outline of Final Research Achievements

The phage display is an excellent method to discover peptide sequences that bind to target molecules. However, it has been challenging to apply this method to membrane proteins that are difficult to be isolated and purified. Therefore, the author attempted to obtain peptide ligands selective for target proteins expressed on the cell surface by chemically modifying phage with Ni complexes. First, to establish the chemical modification reaction on phage, the reaction conditions were elucidated using the fluorescent dye NBD. Based on these results, the author modified the phage with Ni complexes and performed selection of peptide ligands for His-tag fused hDM2. The results showed that the modification of Ni complexes significantly enriched the phage to bind the target protein.

Free Research Field

生物有機化学

Academic Significance and Societal Importance of the Research Achievements

ファージディスプレイ法は、中分子医薬品開発の実績のある基盤技術である。しかし、リガンド探索において、バイアスが強い点や膜蛋白質に適用できない点など致命的な欠点も多い。そのため、これらを解決したリガンド探索法は、創薬の革新的技術となり得る。本研究で開発した手法は効率的にペプチドリガンドを探索でき、次世代の創薬技術の前進的な手法である。本手法の発展と更なる改良によって、より実践的な中分子リガンド探索法に進化すると期待できる。