Development of a high-throughput screening for nerve agent detoxifying materials using a fully-automated robot-assisted biological assay.
Developing improved medical countermeasures against chemical warfare agents (nerve agents) is urgently needed but time-consuming and costly. Here we introduce a robot-assisted liquid handling system with warming, cooling and incubating facilities to screen the detoxifying properties of biological and chemical materials against nerve agents. Two biological tests were established and plasma from various species, DFPase and three cyclodextrins were used as test materials. In test 1, plasma was mixed with sarin or VX and the inhibitory potency of the incubate was determined with human acetylcholinesterase (AChE) at 0, 30 and 60 min. In test 2, test materials and nerve agents were mixed and incubated. Between 0 and 40 min samples were taken and incubated for 3 min with AChE and the residual AChE inhibition was determined to enable the semi-quantitative evaluation of the detoxification kinetics. The automated assays proved to be highly reproducible. It was possible to pre-select detoxifying reagents with test 1 and to determine more detailed detoxifying kinetics with test 2. In conclusion, the automated assay may be considered as a versatile tool for the high-throughput screening of potential detoxifying materials against different nerve agents. With this two-step assay it is possible to screen effectively for detoxifying materials in a high-throughput system.
Wille
T
T
Bundeswehr Institute of Pharmacology and Toxicology, Neuherbergstrasse 11, 80937 Munich, Germany.
Thiermann
H
H
Worek
F
F
D000818:Animals; D001681:Biological Assay / Q000295:instrumentation / Q000379:methods*; D054045:Biological Warfare Agents; D015415:Biomarkers; D002801:Cholinesterase Reactivators / Q000494:pharmacology*; D003505:Cyclodextrins / Q000494:pharmacology; D004353:Drug Evaluation, Preclinical / Q000295:instrumentation / Q000379:methods*; D004912:Erythrocytes / Q000187:drug effects; D006168:Guinea Pigs; D006801:Humans; D066298:In Vitro Techniques; D008253:Macaca mulatta; D009946:Organothiophosphorus Compounds / Q000097:blood / Q000633:toxicity; D044345:Phosphoric Triester Hydrolases / Q000097:blood; D011817:Rabbits; D051381:Rats; D012371:Robotics; D012524:Sarin / Q000097:blood / Q000633:toxicity; D012984:Software; D013045:Species Specificity; D013552:Swine
D054045:Biological Warfare Agents; D015415:Biomarkers; D002801:Cholinesterase Reactivators; D003505:Cyclodextrins; D009946:Organothiophosphorus Compounds; D044345:Phosphoric Triester Hydrolases; D012524:Sarin; C009680:VX-agent; C020061:diisopropyl-fluorophosphatase