| 학회 | 한국화학공학회 |
| 학술대회 | 2017년 봄 (04/26 ~ 04/28, ICC 제주) |
| 권호 | 23권 1호, p.432 |
| 발표분야 | 생물화공 |
| 제목 | Time-resolved sample preparation for cryo-EM structural analysis of protein complex by microfluidic mixing and spraying technique |
| 초록 | Determining the intermediate structures of biomolecular assemblies is the very first step in developing a new drug in many cases. Cryo electron microscopy (cryo-EM) techniques have provided an increasing number of structures at atomic or near-atomic resolution of both soluble and membrane-integrated biomacromolecules, many of which have been widely considered almost impossible to achieve by X-ray crystallography or NMR. However, it is still challenging to discover the structural changes of protein assemblies that usually take place less than a second. Here we have developed a cryo-EM sample preparation method for analyzing structures of protein assemblies with microfluidic platform that has efficient mixing channel and spraying domain to capture the protein complex intermediate with defined time. We confirmed that mixing efficiency of two fluorescence were above 90% after the seventh junction and set the number of junction to seven. To capture the protein solutions onto TEM grid, we used electrospray technique with high voltage of 8 kV and confirmed that proteins remain intact by conducting β-galactosidase enzymatic assay. |
| 저자 | 이병진1, Enchev Radoslav Ivanov2, Sung Sik Lee2, Matthias Peter2, 이창수1 |
| 소속 | 1충남대, 2Institute of Biochemistry |
| 키워드 | 생물화공 |
| 원문파일 | 초록 보기 |
