Analytical Chemistry

National Center For Advancing Translational Sciences

Investigators

Linked publications, trials & patents

Abstract

The COVID-19 global pandemic has continued to have an impact on ACC research and support efforts due to modified laboratory operating procedures, as well as a shift to a hybrid work environment. We are still navigating the logistical challenges of providing core analytical chemistry services while maintaining the health and safety of DPI staff. The success of DPI research projects, initiatives, and programs depends on the uninterrupted operation of analytical services and resources, which facilitates forward progress and translational discoveries. Analytical data and analysis must be made promptly available in a convenient format to allow timely and informed decisions regarding the direction or even continuation of projects. To accomplish this, the ACC developed and modified protocols and workflows to handle the increased workload. These efforts have allowed the ACC to maintain involvement in COVID-related projects while also restarting research projects halted during the pandemic, as well as initiate new projects. NCATS has become a key partner in the Antiviral Program for Pandemics (APP), a multi-agency initiative that includes NIAID and BARDA, to accelerate antiviral development through early discovery and preclinical development of safe and effective oral antivirals. The initial priority for the APP is to develop treatments for SARS-CoV-2 and other coronaviruses, with the program expanding to address other virus families with pandemic potential. NCATS will apply its proficiency in drug discovery and development and its cutting-edge technologies to fill critical needs of the APP, such as (i) target validation, (ii) high-throughput screening (HTS) for drug starting points, (iii) medicinal chemistry for lead optimization, and (iv) preclinical, investigational new drug (IND)-enabling development. APP chemists within DPI are tasked with the discovery and development potential small molecule antiviral treatments for a range of viruses that have pandemic potential. In order to meet the increased demand for sample purification and analysis, the ACC needed to improve throughput and expand capabilities through procurement of additional instrumentation and equipment, as well as the application of new technologies. The ACC continues to provide core analytical chemistry services involving the isolation, purification, identification, and analysis of therapeutic modalities for targeted disease states. We are utilizing the Sample Management and Resource Tracking (SMART) Centralized Sample Purification and Processing Platform for the transfer of compounds from Medicinal Chemistry to Compound Management (CoMa) enabling faster turnaround between small molecule synthesis and biological assays. The purification 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. While these efforts have been highly successful over the past 10 years, the high-performance liquid chromatography (HPLC) instrumentation has seen heavy usage. Therefore, we have procured a replacement preparative purification system, as well as a UPLC-MS system for faster and more efficient sample analysis. To improve upon our sample purification and processing workflow, we have been developing an automated purification platform that will operate as a stand-alone system or an integrated module within the automated laboratory. This innovative system consists of a custom liquid handling platform that incorporates analytical and purification LC instruments into a single automated unit. This will enable replacement of manual steps in workflow with automated protocols as a means of accelerating the drug discovery process. A key aspect of ACC research is the utilization of LCMS analysis throughout all stages of the workflow for reaction monitoring, identity verification, and purity analysis of the desired compounds. To improve efficiency and increase productivity, the ACC has procured the Lab2Lab pneumatic sample transport system, which sends 2D-barcoded minitubes between labs, storage systems, and analytical instrumentation. Chemists located in five labs across two buildings will be able to submit samples directly from their bench for transport to centrally located LCMS instruments near the Analytical Chemistry lab space where analytical measurements are performed. This enables ease of oversight and maintenance, sample tracking capabilities and maximization of instrument usage, and freeing of limited laboratory space. The Lab2Lab system aims to tackle the growing needs of analytical chemistry by streamlining analytical workflows of not just LCMS but also nuclear magnetic resonance (NMR) spectroscopy. The ACC has acquired a Bruker 400 MHz NMR along with the Lab2NMR platform to enable automated NMR operations. The NMR spectrometer outfitted with the Lab2NMR platform is connected to the Lab2Lab system for the transport of vials for automated NMR sample preparation and data acquisition. The Lab2NMR ensures the ACC can seamlessly integrate automated NMR data acquisition and analysis into our workflow and processes. NMR analytical capabilities continue to expand within the ACC through the acquisition of advanced instrumentation and the incorporation of new and emerging techniques in order to detect, identify, quantify, and validate a wide array of therapeutic modalities. We are in the process of replacing aging instrumentation with obsolete technology that has experienced increased malfunctioning as the systems approach their end of life. We have purchased a JEOL 400 MHz NMR for routine open access to the medicinal chemists. The old NMR spectrometer was decommissioned to make space for the new instrumentation. To aid in the discovery phase of drug development, we have been developing our fragment-based screening (FBS) program over several years. NMR-based fragment screening is a powerful approach due its ability to detect very weak ligand-protein binding affinities across a large and diverse chemical space. In anticipation of onboarding a growing number of FBS projects, the ACC has acquired a new fragment library with high chemical diversity to replace our current first-generation fragment library. In collaboration with the ASPIRE program, the ACC continues to develop our automated quantitative NMR (qNMR) platform for concentration determination, purity determination, and reaction analysis. For mass spectrometry (MS), the ACC in collaboration with the Informatics Core (IFX) has been developing a multi-omics analysis platform that will include, but is not limited to proteomics, metabolomics, and lipidomics. Current MS instrumentation in the ACC is primarily configured for routine targeted MS-based studies. We have recently purchased an Agilent LC-QTOF system that will be dedicated to untargeted metabolomics experimentation, which will fulfill an unmet research need at NCATS. Untargeted metabolomic profiling captures a detailed view of the chemical processes underlying cells and is the closest representation of a given phenotype (compared to other omics). Several applications of this instrument include (i) identifying biomarkers of toxicity, treatment response, and diseases, (ii) elucidating biological mechanisms underlying cellular differentiation, disease development, and treatment response, (iii) narrowing down possible compounds to evaluate in assays based on metabolism features. The LC-QTOF system allows the ACC and IFX to expand multi-omic capabilities of NCATS.