Immunochemical characterization of anti-acetylcholinesterase inhibitory monoclonal antibodies.
Monoclonal antibodies (mAbs) were prepared against native or DFP-inhibited Torpedo californica acetylcholinesterase and native or DFP-, MEPQ-, and soman-inhibited fetal bovine serum acetylcholinesterase. The cross reactivity of these antibodies with acetylcholinesterases from various species and their ability to inhibit catalytic activity were determined. Eight antibodies were found to inhibit catalytic activity of either Torpedo or fetal bovine serum enzyme. In all cases the antibodies bound to the native form of the enzymes and in some cases even to the denatured form. None of the antibodies recognized human or horse serum butyrylcholinesterase. Sucrose density gradient centrifugation of enzyme-antibody complexes provided two types of profiles, one with multiple peaks, indicating numerous complexes between tetrameric forms of the enzyme, and the other with single peaks, demonstrating complex formation within the tetrameric form. Different antibodies appeared to interact with slightly different regions, but in all cases the binding encompassed the peripheral anionic site. Decrease in catalytic activity of the enzyme was most likely caused by conformational changes in the enzyme molecule resulting from interaction with these mAbs.
Gentry
M K
MK
Division of Biochemistry, Walter Reed Army Institute of Research, Washington, DC 20307-5100.
Saxena
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A
Ashani
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Doctor
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BP
D000110:Acetylcholinesterase / Q000097:blood / Q000737:chemistry / Q000276:immunology*; D000818:Animals; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology*; D002091:Butyrylcholinesterase / Q000097:blood; D002417:Cattle; D002800:Cholinesterase Inhibitors / Q000276:immunology / Q000494:pharmacology*; D003429:Cross Reactions; D000939:Epitopes / Q000378:metabolism; D006041:Goats; D006736:Horses; D006801:Humans; D007114:Immunization; D007120:Immunochemistry; D007531:Isoflurophate / Q000378:metabolism; D051379:Mice; D011487:Protein Conformation; D011817:Rabbits; D012756:Sheep; D013552:Swine; D014101:Torpedo / Q000378:metabolism; D014316:Tritium
D000911:Antibodies, Monoclonal; D002800:Cholinesterase Inhibitors; D000939:Epitopes; D014316:Tritium; D007531:Isoflurophate; D000110:Acetylcholinesterase; D002091:Butyrylcholinesterase