Enzyme immunoassays for leukotrienes C4 and E4 using acetylcholinesterase

Philippe Pradelles, Catherine Antoine, Jean Paul Lellouche, Jacques Maclouf

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Abstract

The chapter presents a study on enzyme immunoassays for leukotrienes C4 and E4 using acetylcholinesterase. The chapter discusses successfully developing acetylcholinesterase from Electrophorus electricus as a label for various eicosanoids, thereby providing an enzyme immunoassay with sensitivities equal to or superior to those achieved with 125I radioactive tracers. A similar approach for LTC4 and LTE4 is undertaken. However, because of specific problems inherent with these molecules, a dual strategy is combined to prepare the protein-LT, conjugates necessary for the generation of antibodies and corresponding enzyme tracers. The chapter describes the development of such assays. The chapter discusses developing enzyme immunoassays for peptido-LTs by combining separate approaches to attach these haptens to macromolecules. The final strategy employed in labeling the LTs with the enzyme was arrived at as a compromise between the efficiency of the coupling reaction and preservation of the enzymatic activity. Although LTC4 and LTE4 may not be the relevant metabolites to evaluate the in vivo production of peptido LTs, there are a number of in vitro situations, when such assays are needed. In most systems, isolated cells only generate LTC4, whereas in all other in vitro situations, the overall synthesis of peptido LTs can easily be estimated after enzymatic transformation of all peptido-LTs into LTE4.

Original languageEnglish
Pages (from-to)82-89
Number of pages8
JournalMethods in Enzymology
Volume187
Issue numberC
DOIs
StatePublished - 1 Jan 1990

Bibliographical note

Funding Information:
These studies were made possible by financial support from Commissariat Atomique and grants Srom CNAMTS/INSERM and CNRS.

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