Simultaneous Determination of α-Fetoprotein and Free β-Human Chorionic Gonadotropin by Element-Tagged Immunoassay with Detection by Inductively Coupled Plasma Mass Spectrometry

Abstract

Abstract Background: An inductively coupled plasma mass spectrometry (ICP-MS)-based immunoassay has been proposed independently by Baranov et al. (Anal Chem 2002;74:1629–36) and our group, but the applicability of this method for multianalyte analysis in clinical samples has not been fully illustrated. We developed a dual-label immunoassay method for the simultaneous determination of α-fetoprotein (AFP) and free β-human chorionic gonadotropin (hCGβ) in human serum. Methods: Monoclonal antibodies immobilized on microtiter plates captured AFP and hCGβ, which were detected by use of Eu3+-labeled anti-AFP and Sm3+-labeled anti-hCGβ monoclonal antibodies. Eu3+ and Sm3+ were dissociated from the immunocomplex with HNO3 solution (10 mL/L) and delivered by peristaltic pump to the ICP mass spectrometer. Results: The measurable ranges of AFP and hCGβ were 4.6–500 and 5.0–170 μg/L, respectively, with detection limits of 1.2 and 1.7 μg/L (3 SD above mean of zero calibrator), respectively. The intraassay imprecision (CV) for AFP was 8.3%, 4.0%, and 2.7% at 16.3, 86, and 354 μg/L, respectively, and the interassay CV was 10%, 5.7%, and 3.5%. For hCGβ, the intraassay CV was 5.4%, 6.4%, and 3.1%, respectively, at 10.5, 45.2, and 105 μg/L, and the interassay CV was 7.2%, 8.0%, and 3.7%. Comparison with IRMAs for AFP and hCGβ yielded correlation coefficients (r2) of 0.97 and 0.95. Conclusions: Two proteins can be measured simultaneously by immunoassays using two rare earth elemental tags (Eu3+ and Sm3+) and ICP-MS detection. The multielement capability and the multiple potential elemental labels make ICP-MS attractive for multianalyte immunoassays. Implementation of ICP-MS-linked immunoassays may be relatively straightforward because the labeling and immunoreaction procedures have been well developed for clinical time-resolved immunofluorometric assays.