Acridinium esters as high-specific-activity labels in immunoassay.

Abstract

A chemiluminescent acridinium ester has been synthesized that reacts spontaneously with proteins to yield stable, immunoreactive derivatives of high specific activity. The compound has been used to prepare chemiluminescent monoclonal antibodies to human alpha 1-fetoprotein having average incorporation ratios as great as 2.8 mol of label per mole of antibody, which corresponds to a detection limit of approximately 8 X 10(-19) mol. These antibodies have been used in the preliminary development of a two-site immunochemiluminometric assay for human alpha 1-fetoprotein, which requires only a 30-min incubation and a quantification time of 5 s per sample.

763 Comparison between MAGIC LITE-and CAP-system: Two specific IgE assays

“…In brief, the allergen solid phase (250//I) is washed, and 25 /(I of either the test serum, the standards or the control serum are added and the mixture is incubated at room temperature (approximately 20 C) for 30 min. After another washing procedure, a specific anti-human IgE monoclonal antibody labelled with acridinium ester as the chemiluminescent compound [13] is added and incubated at room temperature, then washed (wo limes. Demineralized water is added and the lighl reactions of the samples are detected in a chemiluminometer (Ciba Corning, Germany) by automatic addition of an oxidating reagent (H2O2) and determination of photocmmis-sionat426nm [14].…”

Nucleophilic addition to the 9 position of 9‐phenylcarboxylate‐10‐methylacridinium protects against hydrolysis of the ester

“…AE-labelled DNA probes were prepared by reacting the N-hydroxysuccinimide acridinium ester (Weeks et al, 1983) with a synthetic oligomer containing a primary alkyl amine inserted into the DNA probe during chemical synthesis. The labelled probes were purified by HPLC.…”

“…An acridinium ester has been shown to be a particularly useful label in biological assays and has been used for both antibody-based (Weeks et al, 1983), and DNA probe-based assays (Arnold et al, 1989). It provides an alternative to radioisotopic detection systems with a longer shelf-life, improved sensitivity, and no need for the special handling required by radioisotopes.…”

Detection Methods Using Chemiluminescence

“…Acridinium esters (AE) are direct chemiluminescent labels for antibodies (Weeks et al, 1983) and DNA probes (Septak, 1989;Nelson and Kacian, 1990;Nelson et al, 1992), in contrast to dioxetane and luminol systems, in which an enzyme label catalyzes the chemiluminescent reaction. N-Methyl acridinium esters react with hydrogen peroxide under basic conditions to yield an excited state N-methylacridone which emits light at 430 nm (reviewed by Nelson and Kacian, 1990).…”