W0079

Parallel High Throughput Screening Method for Protein Crystallization in 96 Well Microplates. Lajos Nyársik^1,2, Hans Lehrach^1,2, Günther Knebel³, ¹ Max-Planck-Institute for Molecular Genetics, Ihnestr. 73, D-14195 Berlin, ² The Protein Structure Factory, Heubnerweg 6, D-14059 Berlin, ³ Greiner Bio-One GmbH, Maybachstr. 2, D-72636 Frickenhausen, nyarsik@molgen.mpg.de.

We have developed a new parallel screening technique for automated protein crystallization in 96 well microplates. The method allows simultaneous sitting-drop and hanging-drop vapor diffusion screening for the determination of reliable crystallization conditions of proteins. A new crystallization microplate assembly (CrystalQuick™) consisting of both a plate and a cover lid was designed to run five parallel crystallization sets in a standard 96 well microplate format. The drops are positioned in preformed microwells with an accurate geometry. The use of microwells enables cost-effective high throughput screening, robotic manipulation and automated unattended detection of the crystallization plates.

The crystallization project of the Protein Structure Factory in Berlin accomplishes high throughput crystallization screening experiments with highly purified proteins and delivers single crystals for X-ray diffraction studies at synchrotron beam lines. The main part of our automated protein crystallization system incorporates liquid handling for rapid plate dispensing, a storage and retrieval system and an automated detection station. All units are connected to a fully computer controlled information and management system. The plates are stored in a huge plate hotel, which is connected to a camera based detection system. Each well of a crystallization microplate assembly is inspected in regular time intervals for the presence of crystals. The parallel screening in microwells enables an effective adoption of robotics for crystallization trials.