The plug-based nanovolume Microcapillary Protein Crystallization System (MPCS).

Publication Type:

Journal Article

Authors:

Gerdts, Cory J; Elliott, Mark; Lovell, Scott; Mixon, Mark B; Napuli, Alberto J; Staker, Bart L; Nollert, Peter; Stewart, Lance

Source:

Acta Crystallogr D Biol Crystallogr, Volume 64, Issue Pt 11, p.1116-22 (2008)

Keywords:

Animals, Burkholderia pseudomallei, Chickens, Crystallization, Crystallography, X-Ray, Micro-Electrical-Mechanical Systems, Microfluidic Analytical Techniques, Microfluidics, Molecular Structure, Muramidase, Oxidoreductases Acting on Sulfur Group Donors

Abstract:

<p>The Microcapillary Protein Crystallization System (MPCS) embodies a new semi-automated plug-based crystallization technology which enables nanolitre-volume screening of crystallization conditions in a plasticware format that allows crystals to be easily removed for traditional cryoprotection and X-ray diffraction data collection. Protein crystals grown in these plastic devices can be directly subjected to in situ X-ray diffraction studies. The MPCS integrates the formulation of crystallization cocktails with the preparation of the crystallization experiments. Within microfluidic Teflon tubing or the microfluidic circuitry of a plastic CrystalCard, approximately 10-20 nl volume droplets are generated, each representing a microbatch-style crystallization experiment with a different chemical composition. The entire protein sample is utilized in crystallization experiments. Sparse-matrix screening and chemical gradient screening can be combined in one comprehensive ;hybrid' crystallization trial. The technology lends itself well to optimization by high-granularity gradient screening using optimization reagents such as precipitation agents, ligands or cryoprotectants.</p>