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Development of an ICP-MS immunoassay for the detection of anti-erythropoietin antibodies
Authors:Yanyan Lu  Zhi Xing  Po Cao  Xinrong Zhang
Institution:a Department of Chemistry, Key Laboratory for Atomic and Molecular Nanosciences of the Education Ministry, Tsinghua University, Beijing 100084, PR China
b Dongzhimen Hospital affiliated to Beijing University of Chinese Medicine, Beijing 100700, PR China
Abstract:A sandwich-type immunoassay linked with inductively coupled plasma mass spectrometry (ICP-MS) has been developed for the detection of anti-erythropoietin antibodies (anti-EPO Abs). Recombinant human erythropoietin (rhEPO) was immobilized on the solid phase to capture anti-rhEPO Abs specifically. After the immunoreactions with Au-labeled goat-anti-rabbit IgG, a diluted HNO3 (2%) was used to dissociate Au nanoparticles which was then introduced to the ICP-MS for measurements. Under the optimized conditions, the calibration graph for anti-EPO Abs was linear in the range of 35.6-500 ng mL−1 with a detection limit of 10.7 ng mL−1 (3σ, n = 9). The relative standard deviation (R.S.D.) for three replicate measurements of 30.9 ng mL−1 of anti-EPO Abs was 8.43%. The recoveries of anti-EPO Abs in sera at the spiking level of 50, 100, 150, 200 and 400 ng mL−1 were 99.2%, 101.5%, 95.0%, 94.0% and 102.9%, respectively. For the real sample analysis, 26 samples from healthy people and 53 samples from patients with rhEPO treatments were studied. One sample from patients showed significantly higher anti-EPO Abs from other samples, indicating a possibility of immune response of this patient.
Keywords:rhEPO  Anti-EPO antibody  Immunoassay  Inductively coupled plasma mass spectrometry
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