A novel microchip electrophoresis-based chemiluminescence immunoassay for the detection of alpha-fetoprotein in human serum.

A sensitive immunoassay method based on microchip electrophoresis chemiluminescence (MCE-CL) detection technology was developed for the detection of tumor marker alpha-fetoprotein (AFP). This method adopts the non-competitive immunoassay mode, and was conducted after AFP reacted with excessive horseradish peroxidase (HRP) labeled monoclonal antibody. The extreme pH value was adopted in the electrophoresis buffer solution. The use of brij 35 as an additive of electrophoresis buffer increased dramatically the resolution (Rs) and the reproducibility of the analysis. Under the optimized experimental conditions, effective separation of the immune complex Ag-Ab* and free Ab* was achieved within 60s. The peak height of the immune complex Ag-Ab* was taken as quantification of AFP. Good linearity was observed within AFP concentrations ranging from 10ng/mL to 150ng/mL, and the detection limit was found to be 7.2ng/mL (1.0×10-10M). The present method was successfully applied for the detection of AFP in human serum from both healthy and cancer patients, and the AFP levels in the both were found be in the range of 16.5-23.4ng/mL and 416.2-825.4ng/mL, respectively.

[1]  Chen-zhong Li,et al.  Detection and discrimination of alpha-fetoprotein with a label-free electrochemical impedance spectroscopy biosensor array based on lectin functionalized carbon nanotubes. , 2013, Talanta.

[2]  D. S. Hage,et al.  Capillary electrophoresis‐based immunoassays: Principles and quantitative applications , 2008, Electrophoresis.

[3]  Shulin Zhao,et al.  Integrated microfluidic system with chemiluminescence detection for single cell analysis after intracellular labeling. , 2009, Analytical chemistry.

[4]  J. Koenderink.  Q… , 2014, Les noms officiels des communes de Wallonie, de Bruxelles-Capitale et de la communaute germanophone.

[5]  Xiaohong Zhou,et al.  Triple functional DNA-protein conjugates: Signal probes for Pb(2+) using evanescent wave-induced emission. , 2015, Biosensors & bioelectronics.

[6]  Feng Yan,et al.  Automated support-resolution strategy for a one-way chemiluminescent multiplex immunoassay. , 2009, Analytical chemistry.

[7]  Q. Wei,et al.  Facile fabrication of an ultrasensitive sandwich-type electrochemical immunosensor for the quantitative detection of alpha fetoprotein using multifunctional mesoporous silica as platform and label for signal amplification. , 2014, Talanta.

[8]  M. Breadmore,et al.  Polymeric microchip for the simultaneous determination of anions and cations by hydrodynamic injection using a dual-channel sequential injection microchip electrophoresis system. , 2014, Analytical chemistry.

[9]  Huangxian Ju,et al.  Flow-injection chemiluminescent immunoassay for alpha-fetoprotein based on epoxysilane modified glass microbeads. , 2006, Journal of immunological methods.

[10]  Xunjun He,et al.  A highly sensitive and specific electrochemical sensing method for robust detection of Escherichia coli lac Z gene sequence. , 2015, Biosensors & bioelectronics.

[11]  G. G. Stokes.  "J." , 1890, The New Yale Book of Quotations.

[12]  W. Tsai,et al.  Development of a piezoelectric immunosensor for the detection of alpha-fetoprotein , 2005 .

[13]  Xiliang Luo,et al.  Gold nanoparticles and polyethylene glycols functionalized conducting polyaniline nanowires for ultrasensitive and low fouling immunosensing of alpha-fetoprotein. , 2016, Biosensors & bioelectronics.

[14]  James T. Wu,et al.  Serum Alpha Fetoprotein (AFP) Levels in Normal Infants , 1981, Pediatric Research.

[15]  Jie-ke Cheng,et al.  Capillary electrophoretic immunoassay for alpha-fetoprotein with chemiluminescence detection. , 2007, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[16]  M. Kaneko,et al.  Evaluation of alpha-fetoprotein in early infancy. , 1978, Journal of pediatric surgery.

[17]  F. Mostofi,et al.  Detection of AFP and HCG in metastatic testicular cancer after treatment with chemotherapy or radiation therapy. , 1989, Urology.

[18]  Huangxian Ju,et al.  Layer-by-layer hydroxymethyl ferrocene modified sensor for one-step flow/stop-flow injection amperometric immunoassay of alpha-fetoprotein. , 2007, Biosensors & bioelectronics.

[19]  A. Riskin,et al.  Alpha-fetoprotein in the early neonatal period--a large study and review of the literature. , 2004, Clinica chimica acta; international journal of clinical chemistry.

[20]  Shusheng Zhang,et al.  Capillary electrophoresis enzyme immunoassay for alpha-fetoprotein and thyroxine in human serum with electrochemical detection. , 2008, Journal of separation science.

[21]  L. Hongling,et al.  Polydopamine and graphene oxide synergistically modified Prussian blue electrochemical immunosensor for the detection of alpha-fetoprotein with enhanced stability and sensibility , 2015 .

[22]  Non-competitive immunoassay for alpha-fetoprotein using micellar electrokinetic capillary chromatography and laser-induced fluorescence detection. , 2002, Journal of chromatography. B, Analytical technologies in the biomedical and life sciences.

[23]  Uwe Karst,et al.  Recent developments in optical detection methods for microchip separations , 2006, Analytical and bioanalytical chemistry.

[24]  Shulin Zhao,et al.  Quantification of biogenic amines by microchip electrophoresis with chemiluminescence detection. , 2009, Journal of chromatography. A.

[25]  C. G. Bergstrand.  Alphafetoprotein in Paediatrics , 1986, Acta paediatrica Scandinavica.