The high-speed Hydra-Plus-One system for automated high-throughput protein crystallography

Abstract

An automated high-throughput dispenser has been developed for the setup of protein crystallization trials by vapor diffusion or Microbatch methods. The Hydra-Plus-One is composed of a Hydra-PP system equipped with a motorized XYZ-platform, 96 precision glass syringes and a single-channel microsolenoid dispenser, which transfers 100 nl-50 micro l of protein solution with an accuracy of > 90% at a speed of 60s per 96 wells. Up to 300 micro l of premixed cocktails can be aspirated with the 96-syringe-assembly and dispensed into reservoir and droplet wells within 60s. The Hydra-Plus-One combines high precision, reliability and speed in a cost-effective high-throughput system ideally suited for protein crystallization

Applications of the second virial coefficient: protein crystallization and solubility

“…Parameters screened generally include protein concentration, pH (generally a pH range from 3.0 to 9.0 is evaluated in steps of 0.3 pH units), buffer type, precipitating agent type and concentration, temperature, additive type and concentration. This approach, combined with technological developments such as high-throughput protein-expression/purification and automated crystallographic determination protocols, the use of protein engineering to enhance crystallization lattice contacts and robotic liquid-dispensing/crystallization systems, have accelerated the crystallization and structure determination of thousands of new proteins (Blundell & Mizuguchi, 2000;Burley et al, 1999;Mittl & Grü tter, 2001;Montelione & Anderson, 1999;Teichmann et al, 1999;Christendat et al, 2000;Waldo et al, 1999;Terwilliger, 2000;Abola et al, 2000;Hendrickson, 2000;Derewenda, 2004a,b;Rupp et al, 2002;Krupka et al, 2002;Stevens, 2000). However, in spite of the plethora of new protein structures that have resulted from these technological advances, it is clear that robotics alone is not sufficient for the successful crystallization of a significant number of important proteins.…”

Cleaning protocols for crystallization robots: preventing protease contamination

“…An optional contact syringe dispenser arm for lipidic cubic phase (LCP) setup is also available for the Gryphon model. The prototype of these robots was originally conceived in an academic laboratory (Krupka et al, 2002), but no specifications for a cleaning protocol were published.…”

The TB structural genomics consortium crystallization facility: towards automation from protein to electron density

“…Due to the rapid setup, drop sizes down to a total of 500 nl appear reasonably achievable using this technique without need for humidity controlled environment. The Hydra-Innovadyne combination appears to be a fast and relatively inexpensive solution to protein crystallization setup -provided that premixed screens (true random or sparse matrix type) in 96-well format are available (Krupka et al, 2002b). Plates and blocks can readily be loaded, and finished plates automatically transferred to a sealer, with any SBS-standard compliant plate crane if desired.…”