Summary
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1.
Acetylcholinesterase (AChE) was purified 20,000-fold in a 43% yield from 90 g of human cerebellum by combined immunoaffinity and ligand affinity chromatography. The purified enzyme migrated as a 68,000-dalton band during polyacrylamide gel electrophoresis under denaturing and reducing conditions.
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2.
Balb/c mice were immunized with multiple 10-µg injections of this material in order to raise monoclonal antibodies to human brain AChE. Three such antibodies were obtained and characterized.
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Each antibody cross-reacted distinctively with AChEs from other mammals. No antibody recognized human plasma butyrylcholinesterase but all reacted with AChE from human red blood cells.
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Antibodies HR5 and HR3 performed well in two-site immunoassays for AChE. With these assays we compared autopsy samples of cortical region A9 from six controls (nonneurological cases) and five patients with Alzheimer's disease. The latter showed a highly significant 60% deficit of AChE protein.
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The present antibodies will permit additional immunochemical studies of cholinergic systems in dementia.
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Rakonczay, Z., Brimijoin, S. Monoclonal antibodies to human brain acetylcholinesterase: properties and applications. Cell Mol Neurobiol 8, 85–93 (1988). https://doi.org/10.1007/BF00712914
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DOI: https://doi.org/10.1007/BF00712914