and Abstract The X-ray Crystallography and Macromolecular Characterization Shared Resource Facility at the Sidney Kimmel Cancer Center (SKCC), dedicated to supporting cutting edge structural and molecular biology research applied to cancer biology, is one of the 8 National Cancer Institute (NCI)-supported Core Facilities at Thomas Jefferson University (TJU). Because of its potential to enhance existing NIH-funded research programs and further develop programmatic research initiatives at TJU research centers, X-ray crystallography is one of the techniques identified by Dr. Tykocinski, Dean of Jefferson Medical College and Provost of TJU, for strategic development of our research mission. Our existing two crystallization robots, which are 6 and 17 years old, limit our ability to carry out cutting edge structural work. The older robot, a Hydra II Plus One installed in 2001 was decommissioned in Sept 2011 due to constant failure and lack of manufacturing support. The younger unit, an Oryx8 drop setter operational since Feb 2012, is remarkably slow and lacks liquid handling capability, which makes it insufficient for high throughput screening. Therefore, as part of a cost-sharing plan to establish a state-of-the-art center for structural biology at TJU, we request funds to acquire a new NT8 crystallization robot equipped with components dedicated to pipetting viscous solutions in Lipidic Cubic Phase (LCP). We have identified a group of 9 core users from both inside TJU and Temple University who eagerly support this proposal. If funded, TJU will leverage this support with ~$11,500/year to cover the service contract, in addition to funds supporting 40% of a Ph.D.-level facility manager, Dr. Anshul Bhardwaj. The SKCC will also infuse additional funds to cover the remaining 60% of Dr. Bhardwaj salary and running costs. Our preliminary instrument research and tests indicate that the Formulatrix NT8 crystallization robot is ideally suited to meet our needs. This nanoliter-volume liquid handler with drop dispensing capability allows us to identify crystallization hits with great speed and reproducibility starting from microscopic quantities of biological specimen. The NT8 is an ideal fit to meet the high throughput needs of a growing facility. By increasing the efficiency of our protein crystallization pipeline, adding capability to handle membrane proteins, and facilitating automated dispensing of crystallization plates, the proposed instrumentation will significantly enhance the competitiveness of current NIH-funded research projects and expand the body of researchers using our NIH- funded facility, with the ultimate goal of improving the quality of our biomedical research and strengthening the liaison between basic research, cancer research and its clinical applications.
The X-ray Crystallography and Macromolecular Characterization Shared Resource Facility at the Sidney Kimmel Cancer Center, at Thomas Jefferson University is one of the 8 NCI-supported Core Facilities dedicated to developing cutting edge research in structural and molecular biology to support cancer biology research. The current crystallization robots hosted in this core facility are obsolete, slow and have shown an alarming increase in down time and decrease in performance; the result has been reduced or delayed access for our NIH-funded facility users and limitations on our ability to identify crystallization hits for internal and external users. We request matching funds to acquire a new High Throughput Crystallization Robot equipped for Lipidic Cubic Phase (LCP) crystallization. This platform will be maintained as a shared resource, and integrated with the goals of our existing NCI-funded Cancer Center, whose primary mission is to improve public health.
