Abstract
A novel chemiluminescence(CL) substrate-resolved immunoassay platform for the determination of two proteins(NSE and CA15-3) is proposed,based on alkaline phosphatase(ALP) and horseradish peroxidase(HRP) for subsequent CL detection.Several immunoreaction parameters were investigated systematically,including the amount of capture antibody,detection antibody and the dilutions of HRP-SA and anti-FITC-ALP.The dose-response curves showed the linear ranges from 0.5 to 20 ng/mL(R20.99) for NSE and 0.5 to 20 U/mL(R20.99) for CA15-3 with the limits of detection 0.2 ng/mL and 0.2 U/mL,respectively.Serum samples spiked with different amounts of antigen(high、middle、low) were assayed and showed an acceptable spiking recovery.Both the inter-assay and intra-assay variation were less than 10%.Two proteins were determined in a single run and no cross-reaction between assays was found.Overall,this proposed method has been successfully used for the simultaneous determination of dual biomarkers in a single vessel,characteristic of low sample consumption,simple operation with improved assay results and reduced overall cost per assay.This simple and homogeneous technique will find applications in the early diagnosis of human golima.
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