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

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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 (ZnO-NPs) 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.

Schematic of the microplate chemiluminescence sandwich assay for CEA using ZnO-labeled primary antibody and HRP-labeled secondary antibody

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Authors and Affiliations

  1. Biosensor Lab. Department of Chemistry, BITS, Pilani –KK Birla Goa Campus, Goa, 403726, India

    Souvik Pal & Sunil Bhand

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  1. Souvik Pal
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  2. Sunil Bhand
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Correspondence to Sunil Bhand.

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Pal, S., Bhand, S. Zinc oxide nanoparticle-enhanced ultrasensitive chemiluminescence immunoassay for the carcinoma embryonic antigen. Microchim Acta 182, 1643–1651 (2015). https://doi.org/10.1007/s00604-015-1489-5

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  • DOI: https://doi.org/10.1007/s00604-015-1489-5

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