Micro-plate magnetic chemiluminescence immunoassay and its applications in carcinoembryonic antigen analysis

Abstract

A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluorescein isothiocyanate (FITC)-labeled anti-CEA antibodies, CEA antigens, and horseradish peroxidase (HRP)-conjugated anti-CEA antibodies in micro-plate. The immunomagnetic particles coated with anti-FITC antibodies were used as the solid phase for the immunoassay. The separation procedure was carried out by a magnetic plate adaptor and the luminol-hydrogen peroxide (H 2 O 2 )-HRP system was employed for the chemiluminescence detection. The proposed method combined the advantages of the micro-plate reactor and magnetic particle separation technology with the linear range of 5-250 ng mL 1. The detection limit of CEA was 0.61 ng mL 1. The coefficient of the variation was less than 7% and 13% for intra-assay and inter-assay precision, respectively. Compared with the commercial micro-plate chemiluminescent kit, the proposed method showed a good correlation. micro-plate magnetic chemiluminescence immunoassay, carcinoembryonic antigen, tumor marker

Dual‐labeled chemiluminescence enzyme immunoassay for simultaneous measurement of total prostate specific antigen (TPSA) and free prostate specific antigen (FPSA)

“…[22][23][24][25] In the present study, a dual-labeled chemiluminescent enzyme immunoassay (CLEIA) for the simultaneous measurement of TPSA and FPSA was designed, using dual labeling of anti- The fractions containing ALP-labeled IgG were pooled and an equal volume of glycerol was added, mixed, and stored at −20°C for further use.…”

Reprint of "Cathode photoelectrochemical immunoassay based on analyte- induced formation of exciton trapping for carcinoembryonic antigen detection"

“…The average standard deviation gained from 5 independent experiments for all of the 8 concentrations was 2.9%, indicating good reproducibility and reliability of this method. The linear response range was wider than those based on fluorescence resonance energy transfer (0.257-12.9 ng/mL) [35], electrochemical immunoassay (0.01-100 ng/mL) [36], time-resolved fluoroimmunoassay (1-1000 ng/mL) [37], chemiluminescent immunoassay (5.0-250 ng/mL) [38], laser-induced fluorescence (0.7-80 ng/mL) [39], and gold nanoparticle immunoassay (0.10-80 ng/mL) [40] detections. The LOD was also lower than those of electrochemical immunoassay (0.02 ng/mL) [34], time-resolved fluoroimmunoassay (0.5 ng/mL) [37], chemiluminescent immunoassay (0.61 ng/L) [38], and gold nanoparticle immunoassay (0.04 ng/mL) [40] detections.…”

Immunomagnetic assay combined with CdSe/ZnS amplification of chemiluminescence for the detection of abscisic acid

“…The antigen or the antibody was coupled to the beads directly in most of the application of MBs [17][18][19][20][21]. The nonspecific adsorption of other proteins to the MBs was reduced mostly after the proteins (antigen or antibody) were coupled to the MBs.…”

“…Many immunomagnetic assays (IMA) had been reported for proteins [17,18], bacteria [19] and small molecules [20−22], and the combination of ECL with magnetic bead technology for proteins and DNA detection had been mentioned in a recent critical review [23]. A magnetic bead-based DNA hybridization assay has been successfully applied for sequence-specific DNA related to the avian influenza A H1N1 virus in our past work [24].…”