Development of a method for LC-MS analysis for cleared organs

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K14653
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: System genome science
• Research Institution: The University of Tokyo
• Principal Investigator: ODE Koji 東京大学, 大学院医学系研究科(医学部), 助教 (40612122)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: プロテオミクス / 質量分析 / 組織透明化

Outline of Final Research Achievements

We developed a method to acquire local proteomics information from the biological samples subjected to tissue clearing. During the development, we noticed that the reagents used in the tissue-clearing method called CUBIC have following three problems: 1) the reagent(s) inhibits trypsin digestion reaction, 2) it sticks to the capillary of LC-MS system, and 3) it non-specifically binds to the reverse-phase resin used for the desalting peptides. To overcome these problems, we set up a screening system for novel cleansing reagents using purified proteins and cultured cells. We succeeded to find a set of chemicals that can achieve tissue clearing by delipidation and adjust the refractive index, while the chemicals have minimum effect on the protease activity, do not stick to the LC-MS capillaries as well as reverse-phase resin.

Free Research Field

生化学