Radioiodination and Other Labeling Techniques

Grassi, Jacques; Maclouf, Jacques; Pradelles, Philippe

doi:10.1007/978-3-642-71809-0_5

Cited by 8 publications

(1 citation statement)

References 219 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In recent years, our laboratory has developed enzyme immunoassays based on the use of acetylcholinesterase (AChE) from Electrophorus electricus as label (9). We have shown that AChE conjugates of high specific activity can be used to quantify various substances such as eicosanoids (10), substance P (11), atriopeptin (12), or thymulin (13), at sensitivities equal to or greater than those of conventional RIAs using 126I-iodinated tracers.…”

Section: Introductionmentioning

confidence: 99%

Enzyme immunoassays of adenosine cyclic 3',5'-monophosphate and guanosine cyclic 3',5'-monophosphate using acetylcholinesterase

Pradelles¹,

Grassi²,

Chabardès³

et al. 1989

Anal. Chem.

151

View full text Add to dashboard Cite

The pure tetrameric form of acetylcholinesterase (EC 3.1.1.7) from the electric eel Electrophorus electricus has been covalently coupled to 2'-O-succinyl-cAMP tyrosine methyl ester and 2'-O-succinyl-cGMP. Both enzymatic conjugates have been used as tracers in a classical heterogeneous competitive enzyme immunoassay allowing the determination of cAMP and cGMP, respectively. The test was performed in 96-well microtiter plates coated with a mouse monoclonal anti-rabbit Immunoglobulin antibody in order to ensure separation between bound and free moieties of the tracer. Acetylcholinesterase activity bound to the solid phase was measured by colorimetric assay. When standards or samples were first acetylated by treatment with acetic anhydride, the sensitivity of both assays appeared very good since minimum detectable concentration close to 0.04 pmol/mL (2 fmol/well) could be calculated for each assay. Precision was also very satisfying since the coefficient of variation was less than 5% in the 0.2-10 pmol/mL range. Good correlation was noted between enzymoimmunological and radioimmunological measurements of cAMP performed for different biological samples (urine, serum, or tissue extracts).

show abstract

Section: Introductionmentioning

confidence: 99%