title: Nanolitre-scale crystallization using acoustic liquid-transfer technology.
abstract: Focused acoustic energy allows accurate and precise liquid transfer on scales from picolitre to microlitre volumes. This technology was applied in protein crystallization, successfully transferring a diverse set of proteins as well as hundreds of precipitant solutions from custom and commercial crystallization screens and achieving crystallization in drop volumes as small as 20 nl. Only higher concentrations (>50%) of 2-methyl-2,4-pentanediol (MPD) appeared to be systematically problematic in delivery. The acoustic technology was implemented in a workflow, successfully reproducing active crystallization systems and leading to the discovery of crystallization conditions for previously uncharacterized proteins. The technology offers compelling advantages in low-nanolitre crystallization trials by providing significant reagent savings and presenting seamless scalability for those crystals that require larger volume optimization experiments using the same vapor-diffusion format.
authors.affiliation: Department of Discovery Technologies, Roche Palo Alto LLC, 3431 Hillview Avenue, Palo Alto, CA 94304, USA.
authors.forename: Armando G
authors.initials: AG
authors.lastname: Villaseñor
authors.forename: April
authors.initials: A
authors.lastname: Wong
authors.forename: Ada
authors.initials: A
authors.lastname: Shao
authors.forename: Ankur
authors.initials: A
authors.lastname: Garg
authors.forename: Timothy J
authors.initials: TJ
authors.lastname: Donohue
authors.forename: Andreas
authors.initials: A
authors.lastname: Kuglstatter
authors.forename: Seth F
authors.initials: SF
authors.lastname: Harris
mesh_terms: D000162:Acoustics; D000818:Animals; D002645:Chickens; D003460:Crystallization; D018360:Crystallography, X-Ray / Q000379:methods*; D004529:Egg White / Q000737:chemistry; D006018:Glycols / Q000737:chemistry; D054303:HIV Reverse Transcriptase / Q000737:chemistry; D016174:Hepacivirus / Q000378:metabolism; D006801:Humans; D009113:Muramidase / Q000737:chemistry; D053758:Nanoparticles; D036103:Nanotechnology / Q000379:methods; D011505:Protein-Tyrosine Kinases / Q000737:chemistry; D011506:Proteins / Q000737:chemistry; D012709:Serum Albumin / Q000737:chemistry; D014764:Viral Proteins / Q000737:chemistry; D014783:Viscosity
chemical_list: D006018:Glycols; D011506:Proteins; D012709:Serum Albumin; D014764:Viral Proteins; D011505:Protein-Tyrosine Kinases; C080813:emt protein-tyrosine kinase; D054303:HIV Reverse Transcriptase; D009113:Muramidase; C011480:hexylene glycol