In a major advance for chemical defense and public safety, scientists at Lawrence Livermore National Laboratory's (LLNL) Forensic Science Center (FSC) and Purdue University have developed and demonstrated a high-throughput, automated mass spectrometry platform.
Their platform dramatically accelerates the discovery of medical countermeasure candidates against A-series chemical warfare agents, also known as "Novichoks."
The collaborative research, published today in the Proceedings of the National Academy of Sciences (PNAS), provides the first quantitative data on the potency of these agents and identifies promising new directions for antidote development.
A-series nerve agents have gained notoriety in recent years due to their use in high-profile poisonings. Despite the attention they have received, little experimental data exists about their biological effects or how best to treat exposures.
The LLNL-Purdue team addressed this urgent gap by combining expertise in chemical biology, robotics and advanced mass spectrometry to safely and rapidly conduct thousands of potential antidote reactions - reaching throughput rates of up to 7,000 reactions per hour.
"Rapidly responding to emerging chemical threats like the A-series agents requires new approaches to both understand their dangers and develop effective countermeasures," said Brian Mayer, principal investigator and FSC deputy director.
"By leveraging high-throughput mass spectrometry and automated workflows, we can safely generate the critical data needed to identify promising antidote candidates, even for agents that are closely monitored and extremely hazardous."
The team's enabling technology is an automated, label-free mass spectrometry platform that can analyze complex biochemical reactions in seconds, without the need for traditional markers or dyes. This approach allows researchers to directly measure enzyme inhibition and antidote reactivation in authentic human systems, rather than relying on less-representative animal models or computational predictions.
"This inter-institutional, collaborative model brings the world's best tools to the world's toughest problems - safely and at unprecedented speed," said Graham Cooks, a professor at Purdue University and a co-leader of the study.
The three-year LLNL-Purdue study produced three key findings. First, it provided experimental confirmation that A-series agents inhibit human acetylcholinesterase with potencies similar to traditional nerve agents like sarin and VX. Acetylcholinesterase is an enzyme found in the body, some of which is located in the nervous system, that helps to regulate how nerves talk to each other.
Second, it led to the discovery that certain bispyridinium-based oximes, a class of antidotes, can reactivate enzyme function after A-series exposure - challenging previous reports that these agents are untreatable with existing therapies.
Third, it provided a demonstration of a collaborative, multi-institutional workflow for safely generating nerve agent-adducted enzymes, enabling rapid screening at facilities that cannot directly handle chemical warfare agents.
The research underscores the importance of cross-institutional collaboration and technological innovation in national and global security, Mayer said. The automated, joint robotics-mass spectrometry workflow transitioned from Purdue to FSC laboratories not only greatly accelerates the pace of molecular discovery at LLNL, but also dramatically minimizes the risks associated with handling highly toxic substances.
"This approach allows for rapid, large-scale, safe protein experimentation with authentic chemical warfare agents - something that was previously impractical or impossible," said Audrey Williams, the FSC director. "This capability is essential for staying ahead of emerging threats and ensuring that we have effective medical countermeasures ready when they are needed most. It is work that is directly in line with the FSC's mission."
In addition to Mayer and Cooks, other team members include: Nick Morato, a professor at Purdue University and the paper's first author, and Katelyn Mason, Todd Corzett, Saphon Hok, Teneile Alfaro, Carlos Valdez and all FSC staff members.
The work was supported by LLNL's Laboratory Directed Research and Development program and the National Center for Advancing Translational Sciences ASPIRE program, funded through the National Institutes of Health.
