Nanoparticlelabelling-based magnetic immunoassay on chip combined with electrothermal vaporization - inductively coupled plasma mass spectrometry for the determination of carcinoembryonic antigen in human serum

Abstract

A sensitive and selective on chip magnetic immunoassay method, based on a sandwich-type immunoreaction with PbS nanoparticle (NPs) labels in combination with electrothermal vaporization-inductively coupled plasma mass spectrometry (ETV-ICP-MS), was proposed for the determination of carcinoembryonic antigen (CEA). We designed and fabricated a microfluidic chip for magnetic immunoassay, and the prepared iminodiacetic acid modified silica coated magnetic nanoparticles (IDA-SCMNPs) were packed into the central microchannel to form a solid phase column by self-assembly under the magnetic field. After completion of the immunoreaction involving a primary antibody, CEA and a secondary antibody labeled with PbS NPs on a magnetic solid phase packed-column, ETV-ICP-MS was used to determine the concentration of Pb that was released from the captured PbS NPs using an acid-dissolution step. The concentrations of CEA can be correlated with that of Pb. The established method demonstrated a limit of detection of 0.058 μg L⁻¹ for CEA, with a relative standard deviation (RSD) of 6.7% (c = 10 μg L⁻¹, n = 7). A linearity ranging from 0.2 μg L⁻¹ to 50 μg L⁻¹ and a 2-fold enrichment factor (from 60 μL sample solution to 30 μL eluent) were achieved. The proposed method was further validated by analyzing CEA in human serum. The results were in good agreement with those obtained by chemiluminescent immunoassay, which is currently used as a clinical method. Overall, this method offers the advantages of high speed, high sensitivity, high selectivity, low sample/reagents consumption, high integrity and versatility. Moreover, it can be easily applied to other biological and medical assays.