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Methods. 2016 Feb 15;95:70-7. doi: 10.1016/j.ymeth.2015.08.003. Epub 2015 Aug 7.

An integrated pipeline for sample preparation and characterization at the EMBL@PETRA3 synchrotron facilities.

Author information

1
Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany.
2
Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany. Electronic address: r.meijers@embl-hamburg.de.

Abstract

The characterization of macromolecular samples at synchrotrons has traditionally been restricted to direct exposure to X-rays, but beamline automation and diversification of the user community has led to the establishment of complementary characterization facilities off-line. The Sample Preparation and Characterization (SPC) facility at the EMBL@PETRA3 synchrotron provides synchrotron users access to a range of biophysical techniques for preliminary or parallel sample characterization, to optimize sample usage at the beamlines. Here we describe a sample pipeline from bench to beamline, to assist successful structural characterization using small angle X-ray scattering (SAXS) or macromolecular X-ray crystallography (MX). The SPC has developed a range of quality control protocols to assess incoming samples and to suggest optimization protocols. A high-throughput crystallization platform has been adapted to reach a broader user community, to include chemists and biologists that are not experts in structural biology. The SPC in combination with the beamline and computational facilities at EMBL Hamburg provide a full package of integrated facilities for structural biology and can serve as model for implementation of such resources for other infrastructures.

KEYWORDS:

Crystallization; High-throughput; Limited proteolysis; Protein purification; Quality control; Sample optimization

PMID:
26255961
DOI:
10.1016/j.ymeth.2015.08.003
[Indexed for MEDLINE]

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