This initiative seeks to develop an automated piece of instrumentation that can be used to clean previously used microtiter plates, making them suitable for reuse. Given the large quantities of microtiter plates required for high throughput screening any such device developed has the potential of a viable commercial market. Currently in high throughput screening the rule of thumb is to treat every Society for Biomolecular Sciences (SBS) standard assay plate that gets run in a screen as a consumable that gets used only once and is then discarded due to the risk of cross contamination. That being the case, the relationship of assay plates to compound plates to be screened is a 1 to 1 association, meaning as your compound library grows, your demand for assay plates increases, driving costs upwards. Due to this 1 to 1 relationship, for many assays the most expensive part of the screen are the assay plates themselves. In addition to the cost of the plates, these plates are typically made of non-biodegradable plastics (polystyrene, polypropylene, etc.) that will eventually end up in a landfill once discarded. A piece of instrumentation that could utilize some technique to clean used assay plates would allow each plate to be treated as a resource instead of as a consumable, which could greatly reduce screening costs in addition to the amount of solid waste generated. Also, as an automated device this instrument could be used as part of the screening process itself, which would reduce the amount of start up time and system real estate required for plate storage for large scale screens. Another benefit of the instrument being automated is it would remove the need for large scale screens to be run in batches, since one set of assay plates could be used for the entire screen allowing for continuous system operation. Given the potential for cost and environmental savings and the high degree of automated instrumentation used in biological laboratory settings any instrument developed that could allow for the reuse of microtiter plates could potentially have a commercial market. Project Goals The preliminary goal of this project is to develop a functional prototype of an instrument capable of removing both biological reagents and compounds from a used SBS standard assay plate, specifically geared towards biochemical assays. The final product will be an instrument, or set of instruments, that could be integrated as a component of a high throughput screening system in an automated fashion, capable of cleaning plates regardless of the number of sample wells. The long term goal of this project is to bring this instrument to market to meet the needs of those researchers using high quantities of assay plates, for both biochemical and ideally cell based assays. Phase I Activities and Expected Deliverables • Develop a prototype instrument or a detailed plan for a device that meets the following specifications: o Can handle 96, 384, or 1536 plate formats; o Has the ability to utilize multiple potential cleaning solutions while minimizing the need for large quantities of reagent; o Has the capacity to completely dry a cleaned plate; o Does not involve any abrasive touching of the interior of each well, the bottom in particular, that could negatively affect the physical integrity of each well; o Has a maximum clean cycle time of 5 minutes total. • Demonstrates a cleaning process for plates to be used within a biochemical assay with the ability to: o Remove over 99% of a biological reagent such as a BSA solution using different assay detection modes, within the dynamic range of the assay in question (absorbance, luminescence, fluorescence, etc. ) to verify wash results (e.g. an absorbance assay utilizing Bradford staining solution); o Remove over 99% of a chemical compound such as Tannic Acid using different assay detection modes, within the dynamic range of the assay in question to verify wash results (e.g. a luminescence based assay utilizing a dose response curve of Tannic Acid as a control to quantify any residual Tannic Acid left in the sample area of the plate); o Does not degrade assay performance with repeated wash cycles, capable of withstanding up to 50 wash cycles total; o Although not a specific requirement towards Phase I completion, there should be some ability or plan to quantify a Sterility Assurance Level (SAL), geared towards later work in Phase II when some focus is given towards the ability to clean cell based assay plates to ensure there is no contamination between uses. • Cost estimates to manufacture a device capable of meeting the specifications listed above. • Provide NCATS with all data resulting from Phase I Activities and Deliverables. Phase II Activities and Expected Deliverables • Build a prototype instrument that meets the Phase I specifications in addition to several others geared towards the device working as part of a larger automated process: o Is accessible enough to have a plate automatically loaded into the device by standard laboratory robotic equipment; o Has a remote programmatic interface allowing the instrument to be controlled by an external software application through standard laboratory communication protocols (RS-232, TCP/IP, etc.); o Can reliably operate for extended periods of time in an automated fashion (overnight usage with a constant plate throughput limited by the duration of the load/unload time of the device and the cleaning process itself). • Develop detailed procedures to be able to quantify the instruments cleaning effectiveness: o Provide detailed protocols to show the effectiveness of the instrument in removing biological reagents from a used assay plate; o Provide detailed protocols to show the effectiveness of the instrument in removing chemical compounds from a used assay plate; o Develop procedures to potentially allow for the cleaning of cell based assay plates, assuming these plates did not require any additional coating to promote cell adhesion. • Demonstrate the ability of the prototype instrument to run the cleaning procedures as described above in an automated fashion: o A set of assay plates should be run for multiple cycles and show no residual biological or chemical contamination from previous uses; o The assay performance should remain close to constant despite using the same plates repeatedly; o The assay performance should be comparable to using new assay plates; o Multiple assay detection modes should be tested as described in Phase I. • Develop a robust manufacturing plan for the instrument, using off the shelf OEM components where possible to minimize expense. • Provide NCATS with all data resulting from Phase II Activities and Deliverables.