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. 1977 Jul;29(1):16–22.

A radioimmunoassay for the quantitative evaluation of anti-human acetylcholine receptor antibodies in myasthenia gravis.

V M Monnier, B W Fulpius
PMCID: PMC1541026  PMID: 891031

Abstract

A radioimmunoassay was developed for the quantitative evaluation of antibodies to the acetylcholine receptor in the serum of myasthenic patients. AcChR was extracted from human muscle. A detailed preparation of the 125I-labelled alpha-Bgt-AcChR complex used as antigen is reported. Usually, an average of 20 pmol were obtained form 100 g muscle. This preparation is stable for 1 month in presence of an inhibitor of proteolysis and sufficient for performing about fifteen assays. The labelled complex was incubated with increasing amounts of sera and precipitated with anti-human IgG serum. Titres were expressed in pmol 125I-labelled alpha-Bgt-AcChR complex precipitated per ml serum. Out of thirty-nine sera tested thirty-six had positive titres ranging from 0-1-46 pmol/ml. No anti-AcChR were detected in the sera from twenty-seven patients used as controls.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aharonov A., Abramsky O., Tarrab-Hazdai R., Fuchs S. Humoral antibodies to acetylcholine receptor in patients with myasthenia gravis. Lancet. 1975 Aug 23;2(7930):340–342. doi: 10.1016/s0140-6736(75)92779-8. [DOI] [PubMed] [Google Scholar]
  2. Almon R. R., Andrew C. G., Appel S. H. Serum globulin in myasthenia gravis: inhibition of alpha-bungarotoxin binding to acetylcholine receptors. Science. 1974 Oct 4;186(4158):55–57. doi: 10.1126/science.186.4158.55. [DOI] [PubMed] [Google Scholar]
  3. Almon R. R., Appel S. H. Serum acetylcholine-receptor antibodies in myasthenia gravis. Ann N Y Acad Sci. 1976;274:235–243. doi: 10.1111/j.1749-6632.1976.tb47689.x. [DOI] [PubMed] [Google Scholar]
  4. Bender A. N., Ringel S. P., Engel W. K., Daniels M. P., Vogel Z. Myasthenia gravis: a serum factor blocking acetylcholine receptors of the human neuromuscular junction. Lancet. 1975 Mar 15;1(7907):607–609. doi: 10.1016/s0140-6736(75)91886-3. [DOI] [PubMed] [Google Scholar]
  5. Devreotes P. N., Fambrough D. M. Acetylcholine receptor turnover in membranes of developing muscle fibers. J Cell Biol. 1975 May;65(2):335–358. doi: 10.1083/jcb.65.2.335. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. ELLMAN G. L., COURTNEY K. D., ANDRES V., Jr, FEATHER-STONE R. M. A new and rapid colorimetric determination of acetylcholinesterase activity. Biochem Pharmacol. 1961 Jul;7:88–95. doi: 10.1016/0006-2952(61)90145-9. [DOI] [PubMed] [Google Scholar]
  7. Grob D. Introduction: myasthenia gravis in perspective. Ann N Y Acad Sci. 1976;274:1–2. doi: 10.1111/j.1749-6632.1976.tb47670.x. [DOI] [PubMed] [Google Scholar]
  8. Harboe N., Ingild A. Immunization, isolation of immunoglobulins, estimation of antibody titre. Scand J Immunol Suppl. 1973;1:161–164. doi: 10.1111/j.1365-3083.1973.tb03798.x. [DOI] [PubMed] [Google Scholar]
  9. Klett R. P., Fulpius B. W., Cooper D., Smith M., Reich E., Possani L. D. The acetylcholine receptor. I. Purification and characterization of a macromolecule isolated from Electrophorus electricus. J Biol Chem. 1973 Oct 10;248(19):6841–6853. [PubMed] [Google Scholar]
  10. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  11. Lee C. Y., Chang S. L., Kau S. T., Luh S. H. Chromatographic separation of the venom of Bungarus multicinctus and characterization of its components. J Chromatogr. 1972 Oct 5;72(1):71–82. doi: 10.1016/0021-9673(72)80009-8. [DOI] [PubMed] [Google Scholar]
  12. Lindstrom J. M., Lennon V. A., Seybold M. E., Whittingham S. Experimental autoimmune myasthenia gravis and myasthenia gravis: biochemical and immunochemical aspects. Ann N Y Acad Sci. 1976;274:254–274. doi: 10.1111/j.1749-6632.1976.tb47691.x. [DOI] [PubMed] [Google Scholar]
  13. Lindstrom J. M., Seybold M. E., Lennon V. A., Whittingham S., Duane D. D. Antibody to acetylcholine receptor in myasthenia gravis. Prevalence, clinical correlates, and diagnostic value. Neurology. 1976 Nov;26(11):1054–1059. doi: 10.1212/wnl.26.11.1054. [DOI] [PubMed] [Google Scholar]
  14. Meunier J. C., Olsen R. W., Menez A., Fromageot P., Boquet P., Changeux J. P. Some physical properties of the cholinergic receptor protein from Electrophorus electricus revealed by a tritiated alpha-toxin from Naja nigricollis venom. Biochemistry. 1972 Mar 28;11(7):1200–1210. doi: 10.1021/bi00757a014. [DOI] [PubMed] [Google Scholar]

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