A novel microfluidic biosensor based on an electrical detection system for alpha-fetoprotein.

Conventional immunoassays are labor intensive, expensive and time consuming and require large pieces of equipment for detection. Therefore, we have developed and characterized a novel immunoassay methodology comprised of microbeads and microbiochips. In this method, microbeads are used to filter and immobilize antibodies and an immuno-gold silver staining (IGSS) method is then used to amplify electrical signals that correspond to the bound antibodies. The chip used for this system is composed of an inexpensive and biocompatible polydimethylsiloxane (PDMS) layer over a Pyrex glass substrate that contains a platinum (Pt) microelectrode, which is used to detect the electrical signal in this system, the microelectrode is fabricated on the substrate and a microchannel and pillar-type microfilter is formed in the PDMS layer. A sandwich immunoassay approach was applied to detect alpha-fetoprotein (AFP), a cancer biomarker, using this system. The results of this study showed that the time required for a complete assay was reduced by 1h and a detection limit as low as 1 ng/mL was attained when this system used, which indicates that similar bead-based electrical detection systems could be used for the diagnosis of many forms of cancer.

[1]  Il-Hoon Cho,et al.  Plastic ELISA-on-a-chip based on sequential cross-flow chromatography. , 2006, Analytical chemistry.

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

[3]  Zhou Yu,et al.  An immunoassay for ochratoxin A without the mycotoxin , 2007 .

[4]  Feng Yan,et al.  A disposable electrochemical immunosensor for flow injection immunoassay of carcinoembryonic antigen. , 2006, Biosensors & bioelectronics.

[5]  Ching-Lung Lai,et al.  Serological markers of liver cancer. , 2005, Best practice & research. Clinical gastroenterology.

[6]  Cui-Yan Tan,et al.  Colorimetric detection of protein microarrays based on nanogold probe coupled with silver enhancement. , 2004, Journal of immunological methods.

[7]  Raymond T Chung,et al.  Serum alpha-fetoprotein levels in patients with advanced hepatitis C: results from the HALT-C Trial. , 2005, Journal of hepatology.

[8]  L. D. Ciana,et al.  Highly sensitive amperometric enzyme immunoassay for α-fetoprotein in human serum , 1996 .

[9]  Xiulan Sun,et al.  Preparation of gold-labeled antibody probe and its use in immunochromatography assay for detection of aflatoxin B1. , 2005, International journal of food microbiology.

[10]  Takehiko Kitamori,et al.  Microchip‐based immunoassay system with branching multichannels for simultaneous determination of interferon‐γ , 2002, Electrophoresis.

[11]  Kerstin Länge,et al.  Integration of a surface acoustic wave biosensor in a microfluidic polymer chip. , 2006, Biosensors & bioelectronics.

[12]  M. Almeder,et al.  Signal enhancement at the electron microscopic level using Nanogold and gold-based autometallography , 2000, Histochemistry and Cell Biology.

[13]  Yukihiro Ozaki,et al.  Immunoassay using probe-labelling immunogold nanoparticles with silver staining enhancement via surface-enhanced Raman scattering. , 2004, The Analyst.

[14]  C T Lim,et al.  Bead-based microfluidic immunoassays: the next generation. , 2007, Biosensors & bioelectronics.

[15]  H. B. Halsall,et al.  Microfluidic immunosensor systems. , 2005, Biosensors & bioelectronics.

[16]  Jerome P Ferrance,et al.  Microchip‐Based Cell Lysis and DNA Extraction from Sperm Cells for Application to Forensic Analysis , 2006, Journal of forensic sciences.

[17]  Michael Keusgen,et al.  CRP determination based on a novel magnetic biosensor. , 2007, Biosensors & bioelectronics.

[18]  W. Baschong,et al.  Preparation, use, and enlargement of ultrasmall gold particles in immunoelectron microscopy , 1998, Microscopy research and technique.

[19]  Huangxian Ju,et al.  Electrochemical and chemiluminescent immunosensors for tumor markers. , 2005, Biosensors & bioelectronics.