title: An integrated pipeline for sample preparation and characterization at the EMBL@PETRA3 synchrotron facilities.
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.
authors.affiliation: Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany.
authors.forename: Stephane
authors.initials: S
authors.lastname: Boivin
authors.affiliation: Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany.
authors.forename: Sandra
authors.initials: S
authors.lastname: Kozak
authors.affiliation: Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany.
authors.forename: Gry
authors.initials: G
authors.lastname: Rasmussen
authors.affiliation: Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany.
authors.forename: Ioana Maria
authors.initials: IM
authors.lastname: Nemtanu
authors.affiliation: Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Vieira
authors.affiliation: Sample Preparation and Characterization Facility, European Molecular Biology Laboratory (EMBL), Notkestrasse 85, Hamburg 22607, Germany. Electronic address: r.meijers@embl-hamburg.de.
authors.forename: Rob
authors.initials: R
authors.lastname: Meijers
mesh_terms: D018360:Crystallography, X-Ray / Q000592:standards*; D006801:Humans; D046911:Macromolecular Substances / Q000737:chemistry / Q000648:ultrastructure*; D011786:Quality Control; D053838:Scattering, Small Angle; D012984:Software; D013048:Specimen Handling / Q000592:standards; D017356:Synchrotrons / Q000295:instrumentation*; D014961:X-Ray Diffraction / Q000592:standards*
chemical_list: D046911:Macromolecular Substances
keywords: Crystallization; High-throughput; Limited proteolysis; Protein purification; Quality control; Sample optimization