title: From screen to structure with a harvestable microfluidic device.
abstract: Advances in automation have facilitated the widespread adoption of high-throughput vapour-diffusion methods for initial crystallization screening. However, for many proteins, screening thousands of crystallization conditions fails to yield crystals of sufficient quality for structural characterization. Here, the rates of crystal identification for thaumatin, catalase and myoglobin using microfluidic Crystal Former devices and sitting-drop vapour-diffusion plates are compared. It is shown that the Crystal Former results in a greater number of identified initial crystallization conditions compared with vapour diffusion. Furthermore, crystals of thaumatin and lysozyme obtained in the Crystal Former were used directly for structure determination both in situ and upon harvesting and cryocooling. On the basis of these results, a crystallization strategy is proposed that uses multiple methods with distinct kinetic trajectories through the protein phase diagram to increase the output of crystallization pipelines.
authors.affiliation: National Synchrotron Light Source, Brookhaven National Laboratories, Upton, NY 11973, USA.
authors.forename: Vivian
authors.initials: V
authors.lastname: Stojanoff
authors.forename: Jean
authors.initials: J
authors.lastname: Jakoncic
authors.forename: Deena A
authors.initials: DA
authors.lastname: Oren
authors.forename: V
authors.initials: V
authors.lastname: Nagarajan
authors.forename: Jens-Christian Navarro
authors.initials: JC
authors.lastname: Poulsen
authors.forename: Melanie A
authors.initials: MA
authors.lastname: Adams-Cioaba
authors.forename: Terese
authors.initials: T
authors.lastname: Bergfors
authors.forename: Morten O A
authors.initials: MO
authors.lastname: Sommer
mesh_terms: D057205:Automation, Laboratory; D002374:Catalase / Q000032:analysis; D003460:Crystallization; D032426:Marantaceae / Q000737:chemistry; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation* / Q000379:methods; D008958:Models, Molecular; D009211:Myoglobin / Q000032:analysis; D010940:Plant Proteins / Q000032:analysis; D017434:Protein Structure, Tertiary
chemical_list: D009211:Myoglobin; D010940:Plant Proteins; D002374:Catalase