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Pipelines, Robots, Crystals and Biology: What Use High Throughput Solving Structures of Challenging Targets?

[ Vol. 8 , Issue. 2 ]

Author(s):

Christian Kambach   Pages 205 - 217 ( 13 )

Abstract:


With recent advances in the technology and software underlying crystallographic structure solution, demands on both output and functional significance of X-ray structures are soaring. To achieve the required speed and quality also with ever larger and more difficult targets, combining HTP screening methods (robotics based or not) adopted from structural genomics initiatives with thorough expertise and dedicated characterization effort for each individual target is almost a must. I present concepts, practical considerations, and experiences on implementing an HTP technology platform for structural and functional studies on complexes, membrane proteins and other challenging targets. Emphasis lies on the environment of small academic groups engaged exclusively in hypothesis driven projects focused on specific biological systems. Suitability of given HTP protocols for particular target classes, benchmarking and quality control for procedures, and project management issues at the interface between extensive, broad parameter screening and intensive individual target work required by non-SG amenable targets are discussed.

Keywords:

High throughput, screening, structural biology, cloning, expression, structural genomics, crystallography

Affiliation:

Protein Interaction Group,Biomolecular Research, Paul Scherrer Institut, CH5232 Villigen, Switzerland.



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