Analytical Chemistry

National Center For Advancing Translational Sciences

Investigators

Linked publications & trials

Abstract

Over the past year, the ACC successfully maintained resources and provided analytical support for scientists across the 3 DPI scientific branches (ETB, TDB, and CGB). ACC team members worked together to ensure the continuous operability of all service lines and fulfillment of all responsibilities to sustain productivity. We also built and maintained collaborations with internal and external partners for a variety of research projects. The ACC continued to navigate the logistically challenging task of providing core analytical chemistry services to DPI researchers while also conducting our own research initiatives focused on increasing available analytical analysis capabilities with greater throughput in a more efficient and expedient manner. Utilizing the Sample Management and Resource Tracking (SMART) Centralized Sample Purification and Processing Platform, the ACC purified, isolated, identified, and analyzed a wide array of chemical modalities as part of drug discovery efforts. The purification workflow employs semi-preparative liquid chromatography systems utilizing UV- and mass-directed detection and collection to purify a wide range of small molecules and peptides in the milligrams to grams scale. The SMART laboratory information management system (LIMS) continues to be utilized for sample submission, compound purification and processing, sample registration, compound inventory, sample tracking, and data retrieval and management. This process for the rapid progression of compounds from Medicinal Chemistry to Compound Management (CoMa) has an average 5-day cycle time and resulted in >5,000 new compounds synthesized by DPI chemistry being added to the NCATS compound library. Due to the integral role of the SMART LIMS to DPI research programs, the ACC began an upgrade of the platform. The focus is to provide a streamlined application with increased functionality for ready access of analytical and sample data enabling faster and better decision making with overall greater productivity. Additionally, we are working on integrating more and varied instrumentation for broader capture, tracking, and storage of analytical data. We continued to incorporate the Lab2Lab pneumatic tubing system for transport of analytical samples to HPLC-MS and UPLC-MS instruments for centralize daily analytical analysis activities and create more efficient workflows within the chemistry labs. Renovation of the NCATS NMR lab facilities began in January 2025 and the project was successfully completed on time in July 2025 to consolidate DPI NMR spectroscopy operations into a single lab space. The facility includes enough room for up to six NMR systems along with a wet lab for sample preparation, a tank room for cryogen storage, and an office suite allowing ACC staff to be near the instruments for greater oversight. During the construction, the DPI NMR spectroscopy instrumentation was put into storage for protection of the superconducting magnets and system components. The ACC coordinated temporary access to NMR instruments belonging to the Chemistry and Synthesis Center, NHLBI for usage by DPI analytical and medicinal chemists, ensuring the uninterrupted continuation of research projects and programs without the loss of any productivity. We have successfully installed a JEOL 400 MHz NMR and are awaiting the installation of a Bruker 400 MHz NMR connected to a Lab2NMR liquid handling platform for automated sample preparation and analysis. The ACC continued to develop an automated quantitative NMR (qNMR) platform from sample preparation to data acquisition to data analysis. This innovative platform will enable fast and efficient determination of sample concentration, purity analysis, and mixture identification. The technology was used to validate automated NMR sample preparation workflows for sample analysis with an NMR instrumentation from any vendor. We continue to develop an NMR-based fragment screening program as an orthogonal method for discovering and developing chemical probes and biologically active small molecules, which engage with protein targets of interest. This method was used to identify reversible small molecule binders of lecithin-cholesterol acyltransferase (LCAT), an enzyme involved in the clearance of excess cholesterol. The ACC has performed >50,000 sample injections as part of numerous Agilent RapidFire MS-based high-throughput assays providing excellent data that has identified small molecule inhibitors and activators for a number of disease-related protein targets. We continued our development of an Activity-Based Proteomic Profiling (ABPP) platform for kinase selectivity profiling for use by DPI researchers and collaborative partners. This platform was successfully utilized to screen inhibitors of the LATS1/2 proteins for validation of selective target engagement. In addition to determining kinase selectivity, we looked for off-target activity that could contribute to undesirable toxicity from the inhibitors. Furthermore, any discovered off-target proteins presents the opportunity to identify novel small molecule candidates for other therapeutic targets. The ABPP platform is highly adaptable enabling proteomics screening for not just kinases but any class of proteins with a selective probe. We are actively Working to miniaturize the platform for use in a 384-well format to increase sample throughput. We developed an integrated platform for the automated, high-throughput proteomics sample preparation and mass spectrometry (MS) analysis of Caenorhabditis elegans (C. elegans), have long served as a eukaryotic model organism for human biology. The platform incorporates novel Adaptive Focused Acoustics (AFA) technology to extract proteins from C. elegans samples through focused ultrasonication in a temperature-controlled environment. Subsequent optimizations using a Bravo liquid handling system facilitated automated sample preparation process after lysis with the AFA-based system, leading to identification of thousands of proteins from only 3 worms per sample. This platform reduces overall sample preparation time from two days to eight hours and allows simultaneous processing of 384 differential samples – a 30-fold improvement in throughput over traditional methods. Ultimately, this automated and high-throughput platform will allow effective utilization of C. elegans as an orthologous phenotypic model within pre-clinical therapeutic development for a wide range of human diseases. Additionally, the mass spectrometry team has begun to plan for the consolidation of instrumentation into a single lab area for better oversight and management of research activities to enable the sharing of resources within DPI, as well as improving efficiency and increasing productivity of mass spectrometry research efforts. The ACC emphasizes training and mentorship as part of efforts to develop the next generation of translational scientists. Career development of both trainees and staff is a significant priority for the ACC where they will acquire and expand (i) specialized knowledge in their focus area of analytical chemistry, (ii) education in diverse techniques, methods, and procedures, (iii) collaborative experience, and (iv) dissemination of research. It is our goal to provide trainees with directly applicable knowledge and hands-on experience making them outstanding scientists who are highly competitive in the next stages of their career path. A postbaccalaureate IRTA fellow within the ACC who was performing NMR spectroscopy research was accepted into an MD/PhD graduate program and they left the group to begin the next stage of their career. We also extended the appointment of a successful postdoctoral IRTA fellow conducting research focused on mass spectrometry. We onboarded a computer scientist who is upgrading our SMART laboratory information management system (LIMS), as well as working to integrate analytical instrumentation for better data acquisition and management. Staff and trainees are encouraged to annually attend scientific conferences, meetings, and/or workshops specializing in topics pertinent to the NCATS mission. This allows them to learn of the latest advances in analytical chemistry, as well as present their work to the larger scientific community. ACC scientists presented their research at several conferences and meetings virtually and in person where they were authors on 7 posters and 3 presentations. Members of the ACC were co-authors on 8 peer-reviewed publications where we provided experimentation, analysis, and subject matter expertise in the areas of mass spectrometry, high-throughput chemical and biological screening, proteomics, lipidomics, NMR spectroscopy, liquid chromatography, and automated sample preparation.