Screening and optimization of protein crystallization conditions through gradual evaporation using a novel crystallization platform

Sameer Talreja, David Y. Kim, Amir Y. Mirarefi, Charles F. Zukoski, Paul J.A. Kenis

Research output: Contribution to journalArticlepeer-review

Abstract

High-throughput screening of a wide range of different conditions is typically required to obtain X-ray quality crystals of proteins for structure-function studies. The outcomes of individual experiments, i.e. the formation of gels, precipitates, microcrystals, or crystals, guide the search for and optimization of conditions resulting in X-ray diffraction quality crystals. Unfortunately, the protein will remain soluble in a large fraction of the experiments. In this paper, an evaporation-based crystallization platform is reported in which droplets containing protein and precipitant are gradually concentrated through evaporation of solvent until the solvent is completely evaporated. A phase transition is thus ensured for each individual crystallization compartment; hence the number of experiments and the amount of precious protein needed to identify suitable crystallization conditions is reduced. The evaporation-based method also allows for rapid screening of different rates of supersaturation, a parameter known to be important for optimization of crystal growth and quality. The successful implementation of this evaporation-based crystallization platform for identification and especially optimization of crystallization conditions is demonstrated using the model proteins of lysozyme and thaumatin.

Original languageEnglish (US)
Pages (from-to)988-995
Number of pages8
JournalJournal of Applied Crystallography
Volume38
Issue number6
DOIs
StatePublished - Dec 2005

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

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