SBIR-STTR Award

The NIH has committed substantial resources to both traditional and - through its Protein Structure Initiative - high-throughput approaches to protein structure determination, and similar commitments have been made in Europe and Asia. A survey of the results for soluble proteins shows that the main bottlenecks are obtaining high-quality crystals and obtaining high-quality diffraction data for accurate structures. Mitegen, LLC has succeeded in rapidly turning academic discoveries at Cornell into commercial products that are having a major impact on the X-ray diffraction pipeline. Phase I of this project will focus on development of new crystallization platforms based on popular automated drop dispensing technology. Ongoing fundamental and applied studies at Cornell are investigating the physics of protein crystal growth and of the dispensing, dynamics and confinement of protein-containing liquid drops. These studies have led to fundamentally new designs for crystallization plates that promise to reduce storage requirements, allow in-situ X-ray examination of crystal quality, improve equilibration kinetics and the reproducibility of crystallization experiments, and improve the ease with which crystals can be retrieved for X-ray structure determination. Concurrently, tools will be developed for crystal retrieval and X-ray structure determination that integrate with these crystallization platforms and that are suitable for "remote" and automated crystal mounting. These plate and tool designs promise to be highly cost competitive with existing technologies. The Phase I goal is to optimize these designs and begin commercial prototype production. Close collaboration with industrial and academic automation groups (particularly those associated with the PSI) will ensure that the designs are viable. Phase II will focus on more advanced designs incorporating insights from fundamental and applied studies at Cornell, on automated crystal retrieval, and on development of mounts for X-ray microscopies. Relevance: Protein crystallography is a central component of modern structural genomics and drug discovery efforts. By facilitating faster and more efficient protein structure determination, the proposed research will assist in understanding the mechanisms underlying disease and in developing new treatments.

Thesaurus Terms:
Crystallization, High Throughput Technology, Method Development, Protein Structure Function Biomedical Automation, Biophysics, Chemical Kinetics, Functional /Structural Genomics, Telemetry X Ray Crystallography, Bioengineering /Biomedical Engineering

Our understanding of the molecular mechanisms of life is increasingly based upon our knowledge of the three dimensional structure of proteins, nucleic acids, viruses and biomolecular complexes. Structure provides insight into function (or malfunction), and provides a starting point for modern drug discovery and molecular medicine. Biomolecular structures are most often determined using X-ray crystallography of crystallized biomolecules. Over the last decade, high-throughput methods have been introduced that have automated many aspects of protein expression, purification, crystallization and crystallography, and that have lowered the cost per structure determined. However, the growth and harvesting of protein crystals remains a major bottleneck in the pipeline from gene to three-dimensional molecular structure and from structure to pharmaceutical therapy. This Phase II STTR proposal is focused on developing and commercializing improved methods for conventional and high-throughput crystallization and for crystal harvesting and X-ray data collection. An examination at Cornell University of how liquid contact lines interact with surfaces has led to a simple technology for precisely defining the positions of dispensed liquid drops and firmly holding them to those positions, regardless of their chemical composition. This technology forms the basis for a new approach to protein crystallization plates that eliminates the liquid-confining wells of conventional plates. These new plates promise to provide precise control over drop position and shape, resulting in more reproducible crystallization kinetics and simplified image analysis. They will allow hanging and sitting drop growth of soluble and membrane proteins using a single plate, and in situ optical, UV and X-ray analysis with low background. They will meet a critical need for plates optimized for easy X-ray examination of screening and crystallization outcomes and also allow in situ structure determination. They will be compatible with all existing drop dispensing and plate handling hardware, lowering barriers to market entry. This project will continue the scientific and commercial development of these plates and explore other biomedical applications of drop pinning technology. In Phase I we successfully developed and commercialized several new tools for crystal retrieval and X-ray data collection. We will continue this development in Phase II, focusing on improved tools for microcrystallography, for automated sample mounting, and for X-ray beam alignment and energy measurement. Together, these technologies should have significant impact on the productivity of high- throughput structural genomics and drug discovery efforts.

Public Health Relevance:
Our understanding of the molecular mechanisms of life is increasingly based upon our knowledge of the three dimensional structure of proteins, nucleic acids and viruses. Structure provides insight into function (or malfunction), and provides a starting point for modern drug discovery and molecular medicine. This project will develop and commercialize new technologies for use in determining the protein structures by X-ray crystallography that promise to speed up the progression from gene to pharmaceutical therapy.

Thesaurus Terms:
3-D Structure;3-Dimensional Structure;3d Structure;Acceleration;Affect;Applied Research;Applied Science;Area;Asia;Biotechnology;Calibration;Chemicals;Collaborations;Commit;Complex;Crystallization;Crystallographies;Crystallography;Data Collection;Development;Drops;Europe;Evaluation;Filamentous Fungi;Film;General Viruses;Generalized Growth;Genes;Government;Growth;Harvest;Image;Image Analyses;Image Analysis;Imaging Technology;In Situ;Injection Of Therapeutic Agent;Injections;Kinetics;Knowledge;Loinc Axis 2 Property;Life;Liquid Substance;Macromolecular Structure;Marketing;Mass Photometry/Spectrum Analysis;Mass Spectrometry;Mass Spectroscopy;Mass Spectrum;Mass Spectrum Analyses;Mass Spectrum Analysis;Measurement;Membrane Proteins;Membrane-Associated Proteins;Methods;Molds;Molecular;Molecular Medicine;Molecular Structure;Nih;National Institutes Of Health;Nucleic Acids;Optics;Organized By Structure Protein;Outcome;Psi;Pattern;Performance;Persons;Pharmaceutical Agent;Pharmaceuticals;Pharmacologic Substance;Pharmacological Substance;Phase;Position;Positioning Attribute;Process;Production;Productivity;Property;Protein Structure Initiative;Proteins;Proteins [by Structure];Radiation, X-Rays, Gamma-Rays;Research Resources;Resources;Retrieval;Roentgen Rays;Sttr;Sampling;Screening Procedure;Series;Shapes;Shipping;Ships;Single Crystal Diffraction;Small Business Technology Transfer Research;Source;Speed;Speed (Motion);Spottings;Structure;Surface;Surface Proteins;Synchrotrons;Technology;Testing;Tissue Growth;United States National Institutes Of Health;Universities;Virus;Work;X Ray Crystallographies;X Ray Diffraction;X Ray Diffraction Analysis;X-Radiation;X-Ray Crystallography;X-Ray Diffraction;X-Ray Diffraction Crystallography;X-Ray/Neutron Crystallography;X-Rays;X-Rays Radiation;Xray Crystallography;Xray Diffraction;Xrays;Base;Commercialization;Cost;Design;Designing;Developmental;Drug Discovery;Fluid;Gene Product;Image Evaluation;Imaging;Improved;Insight;Liquid;Meetings;New Approaches;New Technology;Novel Approaches;Novel Strategies;Novel Strategy;Novel Technologies;Ontogeny;Optical;Programs;Protein Expression;Protein Structure;Prototype;Response;Screening;Screenings;Structural Genomics;Success;Three Dimensional Structure;Tool