A method of cryoprotection for protein crystallography by using a microfluidic chip and its application for in situ X-ray diffraction measurements

Masatoshi Maeki, Ashtamurthy S. Pawate, Kenichi Yamashita, Masahide Kawamoto, Manabu Tokeshi, Paul J.A. Kenis, Masaya Miyazaki

Research output: Contribution to journalArticlepeer-review

Abstract

We demonstrate a seamless and contactless method from protein crystallization to X-ray analysis using a microfluidic chip with the aim of obtaining a complete crystallographic data set of a protein crystal under cryogenic conditions. Our microfluidics-based approach did not require direct manipulation of the protein crystal. Therefore, the microfluidic chip approach is suitable for novices of X-ray analysis of protein crystals. We also investigated the effect of stepwise cryoprotection on the quality of protein crystals. Protein crystals with cryoprotection via on-chip manipulation did not show deterioration of crystallographic quality of the protein crystal. The complete diffraction data set of a protein crystal, which is required for determining the 3D structure of the target protein, is obtainable by a simple manipulation.

Original languageEnglish (US)
Pages (from-to)4194-4200
Number of pages7
JournalAnalytical Chemistry
Volume87
Issue number8
DOIs
StatePublished - Apr 21 2015

ASJC Scopus subject areas

  • Analytical Chemistry

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