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Acta Crystallogr F Struct Biol Commun. 2019 Feb 1;75(Pt 2):123-131. doi: 10.1107/S2053230X18017338. Epub 2019 Jan 24.

Iterative screen optimization maximizes the efficiency of macromolecular crystallization.

Author information

1
Department of Biochemistry and Cell Biology, Geisel School of Medicine at Dartmouth, Hanover, NH 03755, USA.
2
Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USA.

Abstract

Advances in X-ray crystallography have streamlined the process of determining high-resolution three-dimensional macromolecular structures. However, a rate-limiting step in this process continues to be the generation of crystals that are of sufficient size and quality for subsequent diffraction experiments. Here, iterative screen optimization (ISO), a highly automated process in which the precipitant concentrations of each condition in a crystallization screen are modified based on the results of a prior crystallization experiment, is described. After designing a novel high-throughput crystallization screen to take full advantage of this method, the value of ISO is demonstrated by using it to successfully crystallize a panel of six diverse proteins. The results suggest that ISO is an effective method to obtain macromolecular crystals, particularly for proteins that crystallize under a narrow range of precipitant concentrations.

KEYWORDS:

Sweet16; automated liquid handling; crystallization screening; macromolecular crystallography

PMID:
30713164
PMCID:
PMC6360444
DOI:
10.1107/S2053230X18017338
[Indexed for MEDLINE]
Free PMC Article

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