Zinc oxide nanoparticle-enhanced ultrasensitive chemiluminescence immunoassay for the carcinoma embryonic antigen

Abstract

An ultrasensitive enzyme-linked immunosorbent assay (ELISA) is reported for the determination of carcinoma embryonic antigen (CEA) in human serum. It was realized using a microplate reader using a 384-well plate. Monoclonal antibody (Ab) against CEA (1°Ab) acting as the capture probe was immobilized on zinc oxide nanoparticles (ZnONPs) in the form of self-assembled monolayers (SAMs). CEA captured by 1°Ab was quantified using a sandwich ELISA wherein a polyclonal second antibody against CEA (2°Ab) was used for detection and quantified using an HRP-labeled secondary antibody (3°Ab). The ZnO-NPs-CEA capture probe was deposited on the bottom of the wells in order to enhance capture of CEA. A 3-fold enhancement in the chemiluminescence (CL) signal of luminol is found (compared to a conventional ELISA). CEA can be quantified by this method in concentrations as low as 1 pg·mL −1 . The upper limit of detection is 20 ng·mL −1 . The use of ZnO-NPs also imparts improved thermal stability. When stored at 4°C in phosphate-buffered saline of pH 7.4, the probe displays stability of up to 30 days.

Sol-gel zinc oxide nanoparticles: advances in synthesis and applications

“…They discovered that ZnO-NPs could significantly be used in the systems of cancer diagnosis. Additionally, Pal and Bhand [138] developed the ZnO-NP-based test of ELISA for detecting serum levels of cancer antigens of embryonic. The detection system consisted of a monoclonal carcinoma embryonic antigen-antibody conjugated to ZnO-NP, which formed self-assembled monolayers, and even at 1 pg/ml, smaller quantities of antigen were found [139].…”

A review of enhancers for chemiluminescence enzyme immunoassay

“…This research showed that NaTPB and PPP on CL had a synergistic effect. Pal and Bhand (2015) In the study of bromophenol red and bovine serum albumin (BSA) as the luminol signal co-enhancers in chemiluminescent detection of sequence-specific DNA, the CL signal was increased up to 6×10 4 when bromophenol red and BSA were placed into the luminol-H 2 O 2 -HRP system as the co-enhancers. The results indicated that BSA in the bromophenol red-enhanced CL system showed 36 times stronger CL signals than that without BSA addition.…”

“…Zinc stabilized the luminol radical, increasing the half-life of this radical in the medium). Pal and Bhand (2015) utilized zinc oxide nanoparticle to enhance CL signals of luminol and LOD for the determination of carcinoma embryonic antigen (CEA) in human serum. The CL signal of luminol tripled over the conventional ELISA and the limit of quantitation became as low as 1 pg mL −1 .…”

Preparation of a disposable electrochemiluminescence sensor chip based on an MXene-loaded ruthenium luminescent agent and its application in the detection of carcinoembryonic antigens

“…Currently, with the development of analytical techniques, many methods for CEA detection in real samples have been reported. These methods include fluorescence immunoassay, 6 electrochemical analysis, 7 chemiluminescence immunoassay, 8 and surface plasmon resonance. 9 However, convenient and low-cost methods are still required in clinical diagnosis.…”