The Defense Advanced Research Projects Agency (DARPA) said on Feb. 15 that it is transitioning antibody detection technology to the Defense Department’s Critical Reagents Program. The technology, it said, provides more stability and accuracy in detection of biological toxins and gives the DoD more cost-efficient and more effective bio threat sensors.
The DoD, it said, has been using antibody-based biosensors as its immediate detection tool for dangerous antigens like Anthrax and biological toxins. The biosensors rely on the fact that antibodies bind to antigens, however, sensors have had limitations in to functioning over time and the ability to keep a tight bond between antibody and antigen. The tighter the bond -- or affinity -- between antigen and antibody, the more sensitive a biosensor is over a wider range of threats. Existing DoD biosensors, while effective, had restricted shelf lives, were quickly rendered inoperable by high temperatures and offered limited affinity, it said.
DARPA said it launched the Antibody Technology Program (ATP) in 2009 to address the technological limitations of current antibody-based biosensors. The program set out to achieve revolutionary improvements in the stability of antibodies over time, even in extreme conditions; and control affinity in biosensors to enable detection of numerous antigens by a single unit.
DARPA’s said the ATP ended in 2012 having achieved both goals and with a plan in place to transition the technologies to DoD’s Critical Reagents Program, part of the Joint Program Executive Office – Chemical and Biological Defense (JPE-CBD), for biosensor deployment throughout the military services.
Specifically, DARPA said performers demonstrated the ability to increase antibody temperature stability at 70 degrees Celsius (158 degrees Fahrenheit) to 48 hours, up from the current limit of five to ten minutes. When transitioned to DoD biosensors, the results are projected to eliminate the need for refrigeration while increasing the shelf life by a factor of 36, extending survivability at room temperature (approx. 25 degrees Celsius or 77 degrees Fahrenheit) from one month to approximately three years. DARPA also increased antibody affinity by a factor of 400, thus opening the door to vastly more sensitive, multiplexed biosensors that can test for numerous antigens.
“When you consider the locations of warfighters who have the most potential for biological weapons to be used against them, they are typically environments with extreme temperatures and harsh conditions, and the warfighters themselves are probably operating in small groups,” said Mildred Donlon, the DARPA program manager for ATP. “If it’s going to be useful to these teams, DoD equipment needs to be ruggedized to survive conditions and be easy to use by non-experts. The ATP technology hits these goals.
“By removing temperature stability as a limiting factor, troops will now be able to carry sensors with them without worrying about refrigeration and wondering if the sensor will return an accurate reading,” she said. “According to the Chemical Biological Medical Systems Joint Project Management Office at JPE-CBD, eliminating the need for cold-chain logistics in transport and deployment of sensors is estimated to save DoD in the range of $10 million per year,” Donlon said. “The new stability also means antibodies can be attached to new materials to make potentially more practical sensors to take the place of current beads and strips. Most importantly, by pairing more stable sensors with a huge increase in sensitivity, DARPA is giving troops the confidence to trust the results of what can be literally life-or-death measurements.”
According to DARPA, ATP achieved the results by altering the amino acid sequences within the antibody molecules. Rather than creating an additive stabilizing material, it said, ATP devised methods to make the altered amino acids an integral part of the structure of the antibody molecule.
“Antibody-based biosensors have been in use for roughly 30 years,” Donlon said. “DARPA used recent advances in understanding of protein structure and analysis to determine new ways to alter amino acids, integrate them into an antibody structure, and do so at a sustainable scale.”
DARPA partnered with the U.S. Army’s Edgewood Chemical Biological Center (ECBC) from the beginning of ATP, to first assist with evaluation of performer research proposals, then later in the program to provide ATP performers with unaltered antibodies, conduct testing on the performers’ altered antibodies, and validate results. To ensure that the production methods for modifying antibodies are scalable and cost effective, performers had to submit one-gram samples for testing. The positive results mean that existing DoD antibody stockpiles can be altered to incorporate the new properties of stability and high affinity.
Program performers for ATP included: Affomix Corp. (Branford, CT), purchased by Illumina, Inc. (San Diego, CA.); AnaptysBio, Inc. (San Diego, CA); the Naval Research Laboratory (Washington, D.C.); StableBody Technologies, LLC (Lemont, IL); The University of Texas at Austin (Austin, TX); and the ECBC (Aberdeen, MD), which participated as the validation laboratory. AxioMx, Inc. (Branford, CT) was created to rapidly generate high-quality recombinant antibodies.