Enzyme Immunoassays of Icosanoids Using Acetylcholinesterase

  • P. Pradelles
  • J. Maclouf
  • J. Grassi
Part of the NATO ASI Series book series (NSSA, volume 95)


In the early seventies, the need for the development of sensitive and quantitative assay methods for the analysis of the prostanoids became obvious. Two major complementary methods were selected by the scientists: gas-chromatography combined with mass spectrometry and radioimmunoassay (RIA) techniques. Both methods were improved throughout the years in regard of their specificity and sensitivity. The introduction of deuterated carriers for the first one and the use of low-bleeding fused glass capillary column with high efficiency allowed the detection of a few picograms. The RIA techniques improved their specificity after gradually avoiding the classical pitfalls that result from a mishandling of the assay. The gain in sensitivity was related to the increase of specific radioactivity of the tracers. From the early days of (3H) prostaglandins (PG) with 5–10 Ci/mmole (1) these tracers presently range from 50–200 Ci/mmole. In 1975, we introduced the 125I labeling of these substances after coupling of their carboxyle to the amino group of a potential iodine receptor (e.g. histamine, tyramine or tyrosylmethyl ester) (2). Such an approach allowed us to reach for these tracers the theoretical specific radioactivity of the iodine, i.e. 2,000 Ci/mmole. The gain in sensitivity was x5–10 as compared with the corresponding tritiated systems; further these tracers provided the advantages intrinsic to the iodinated labels (absence of quenching, low cost, rapidity of counting...). More recently, non isotopic immunoassay methods for PG have emerged : they involve enzyme immunoassay (EIA) or chemiluminescence immunoassays. These methods should gain a widespread use due to the reinforcement of the regulation on the use of radioactive substances and because of environmental and pollution requirements. However, until now, these methods are rather disappointing because their sensitivity is still poor (3, 4) especially as compared to iodinated tracers.


Dose Response Curve Chemiluminescence Immunoassay Early Seventy Enzymatic Tracer Unextracted Sample 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. 1.
    Caldwell B.V., Burstein S., Brock W.A. and Speroff L., 1971, Radioimmuno-assay of the F prostaglandins, J. Clin. Endocr., 33: 171.Google Scholar
  2. 2.
    Maclouf J., Pradel M., Pradelles P. and Dray F., 1976, 125I derivatives of prostaglandins. A novel approach in prostaglandin analysis by radioimmuno-assay, Biochim. Biophys. Acta, 431: 139.Google Scholar
  3. 3.
    Hayashi Y., Ueda N., Yokota K., Kawamura S., Ogushi F., Yamamoto Y., Yamamoto S., Nakamura K., Yamashita K., Miyazaki H., Kato K. and Terao S., 1983, Enzyme immuno-assay of thromboxane B2, Biochim. Biophys. Acta., 750: 322.Google Scholar
  4. 4.
    Weerasekera D.A., Koullapis E.N., Kim J.B., Barnard G.J., Collins W.P., Kohen F. and Lindner H.R., 1983, Chemiluminescence immuno-assay of thromboxane B2, in Advances in Prostaglandin, Thromboxane and Leukotriene Research, Vol 11, B. Samuelsson, R. Paoletti and P. Ramwell, Raven Press, New York pp 185.Google Scholar
  5. 5.
    Dray F., B. Charbonnel and J. Maclouf., 1975Google Scholar
  6. Radioimmuno-assay of prostaglandins Fα, E1 and E2 in human plasma, Eur. J. Biochem., 5: 311.Google Scholar
  7. 6.
    Massoulié J. and Bon S., 1976, Affinity chromatography of acetylcholinesterase. The importance of hydrophobic interactions, Eur. J. Biochem., 68: 531.PubMedCrossRefGoogle Scholar
  8. 7.
    Massoulié J. and Bon S., 1982, The molecular forms of cholinesterase and acetylcholinesterase in vertebrates, Ann. Rev. Neurosci., 5: 57.PubMedCrossRefGoogle Scholar
  9. 8.
    Ellman G.L., Courtney K.D., Andres V. and Featherstone R.M., 1961, A new and rapid colorimetric determination of acetycholinesterase activity, Biochem. Pharmacol., 7: 88.Google Scholar
  10. 9.
    Pradelles P., Grassi J. and Maclouf J., 1984, (submitted).Google Scholar
  11. 10.
    Marsh D., Grassi J., Vigny M. and Massoulié J., 1984, An immunological study of rat acetylcholinesterase: comparison with acetylcholinesterases from other vertebrates, J. Neurochem., 43: 204.PubMedCrossRefGoogle Scholar
  12. 11.
    Granström E. and Kindahl H., 1978, Radioimmuno-assay of Prostaglandins and Thromboxane. Adv. Prostaglandin Thromboxane Res., 5: 119.PubMedGoogle Scholar

Copyright information

© Plenum Press, New York 1985

Authors and Affiliations

  • P. Pradelles
    • 1
  • J. Maclouf
    • 2
  • J. Grassi
    • 1
  1. 1.Section de Pharmacologie et ImmunologieCommissariat à l’Energie Atomique C.E.N. SaclayGif/Yvette CèdexFrance
  2. 2.U 150 INSERMLA 334CNRS Hospital LariboisiereParis Cèdex 10France

Personalised recommendations