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Li Z.Y.,CAS Research Center for Eco Environmental Sciences | Zhang Q.Y.,Tsinghua University | Zhao L.X.,CAS Research Center for Eco Environmental Sciences | Li Z.J.,Chemclin | And 3 more authors.
Science China Chemistry | Year: 2010

A micro-plate magnetic chemiluminescence immunoassay was developed for rapid and high throughput detection of carcinoembryonic antigens (CEA) in human sera. This method was based on a sandwich immunoreaction of fluorescein isothiocyanate (FITC)-labeled anti-CEA antibodies, CEA antigens, and horseradish peroxidase (HRP)-conjugated anti-CEA antibodies in micro- plate. The immunomagnetic particles coated with anti-FITC antibodies were used as the solid phase for the immunoassay. The separation procedure was carried out by a magnetic plate adaptor and the luminol-hydrogen peroxide (H2O 2)-HRP system was employed for the chemiluminescence detection. The proposed method combined the advantages of the micro-plate reactor and magnetic particle separation technology with the linear range of 5-250 ng mL-1 The detection limit of CEA was 0.61 ng mL?1. The coefficient of the variation was less than 7% and 13% for intra-assay and inter-assay precision, respectively. Compared with the commercial micro-plate chemiluminescent kit, the proposed method showed a good correlation. © Science China Press and Springer-Verlag Berlin Heidelberg 2010.


Fang L.,Tsinghua University | Fang L.,Yangtze Normal University | Chen H.,Tsinghua University | Ying X.,Chemclin | Lin J.-M.,Tsinghua University
Talanta | Year: 2011

In this work, a micro-plate chemiluminescence enzyme immunoassay by antibody-coated for the determination of aflatoxin B1 (AFB1) in agricultural products has been established. Aflatoxin B1 antibody (AFB1-Ab) was adsorbed physically on polystyrene micro-plate hole as solid phase antibody, which took place immunity-reaction between antigen and antibody with AFB1 standard solution or samples by direct competition. Luminol-hydrogen peroxide chemiluminescence system catalyzed by horseradish peroxidase (HRP) with p-iodophenol enhancement was used as signal detecting system. The effects of several factors, including composition and pH of coating solution, dilution ratio and amount of antibody and enzyme labeled antigen, time of antibody-coating, incubation and chemiluminescence reaction, and other relevant variables upon the immunoasaay were studied and optimized. The linear range of proposed method for AFB1 was 0.05-10.0 ng g-1 with a correlative coefficient of -0.9997. The sensitivity of the proposed method was 0.01 ng g-1. The RSDs of intra- and inter-assay were less than 12.2% and 10.0%, respectively. This method has been successfully applied to the evaluation of AFB1 in agricultural products with recoveries of 79.8%, 101.9% and 115.4% for low, middle and high concentration samples, respectively. It shows a good correlation with the commercial available ELISA kit for AFB1 with correlative coefficient of 0.9098 indicating that the established CLEIA method can be used to determine AFB1 in real samples. © 2011 Elsevier B.V. All rights reserved.


Zhang Q.,Beijing University of Chemical Technology | Zhang Q.,Tsinghua University | Xiao Q.,Tsinghua University | Lin Z.,Tsinghua University | And 3 more authors.
Clinical Biochemistry | Year: 2010

Objective: Glypican-3 (GPC3) is a promising specific tumor maker for hepatocellular carcinoma (HCC). The aim of this study is to establish a method to detect serum GPC3 and evaluate the clinical application on clinical diagnosis. Design and methods: A competitive radioimmunoassay for detecting serum GPC3 was developed. Clinical sera were detected by the proposed method and AFP, CA19-9 chemiluminescence immunoassay kit. Results: The proposed method with high sensitivity, specificity and precision had no or little detectable cross-reactivity with relating tumor markers in the dynamic range from 15 to 500. ng/mL, and the detection limit was 0.5. ng/mL. The level of GPC3 in HCC was obviously higher than that in normal liver or other liver diseases. Additionally, our method showed high shows higher sensitivity and specificity for GPC3 than AFP and combined AFP/CA19-9. Conclusions: This paper provided an applicable competitive radioimmunoassay for GPC3 with high sensitivity, specificity and precision. In addition, using GPC3 for HCC diagnosis was more valuable than AFP. © 2010 The Canadian Society of Clinical Chemists.


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