Approach to quantitative detection of CD146 with the label-free protein biosensor based on imaging ellipsometry

CD146 glycoprotein belonging to cell adhesion molecules is considered to be a novel target on endothelial cell involved in tumor angiogenesis. The biosensor based on imaging ellipsometry (BIE) which is performed in null and off-null mode is used for CD146 detection as a trial by the following steps. Firstly, anti-CD146 antibody as ligand is immobilized on Protein G modified silicon substrate. Then, CD146 test is carried out and its calibration curve is established for the requirement of quantitative detection. Finally, 18 serum samples are detected quantitatively and their results are validated by ELISA's. The sensitivity for CD146 detection achieves the order of ng/ml and the relationship between BIE signal y (grayscale value) and CD146 concentration x (ng/ml) is y=3.3ln(x) +91.3. Compared with ELISA's, the majority of results are in agreement, and the results of two approaches have significant statistic relevance.

[1]  G. Jin,et al.  A label-free multisensing immunosensor based on imaging ellipsometry. , 2003, Analytical chemistry.

[2]  G. Jin,et al.  Development of biosensor based on imaging ellipsometry , 2008 .

[3]  G. Jin,et al.  Silicon surface modification with a mixed silanes layer to immobilize proteins for biosensor with imaging ellipsometry. , 2004, Colloids and surfaces. B, Biointerfaces.

[4]  Peiyu Li,et al.  A novel anti-CD146 monoclonal antibody, AA98, inhibits angiogenesis and tumor growth. , 2003, Blood.

[5]  Gang Jin,et al.  Covalent immobilization of proteins for the biosensor based on imaging ellipsometry. , 2004, Journal of immunological methods.

[6]  J. Johnson,et al.  Expression of MCAM/MUC18 by human melanoma cells leads to increased tumor growth and metastasis. , 1997, Cancer research.

[7]  I. Shih.  The role of CD146 (Mel‐CAM) in biology and pathology , 1999, The Journal of pathology.

[8]  Dongling Yang,et al.  A human SARS-CoV neutralizing antibody against epitope on S2 protein , 2005, Biochemical and Biophysical Research Communications.

[9]  M. Herlyn,et al.  Mel-CAM-specific genetic suppressor elements inhibit melanoma growth and invasion through loss of gap junctional communication , 2001, Oncogene.

[10]  D. Parkin,et al.  Global cancer statistics in the year 2000. , 2001, The Lancet. Oncology.

[11]  Marta Sanchez-Carbayo,et al.  Antibody arrays: technical considerations and clinical applications in cancer. , 2006, Clinical chemistry.

[12]  É. Vivier,et al.  Activation of Human Endothelial Cells via S-Endo-1 Antigen (CD146) Stimulates the Tyrosine Phosphorylation of Focal Adhesion Kinase p125FAK * , 1998, The Journal of Biological Chemistry.

[13]  H. Zhang,et al.  Detection of hepatitis B virus markers using a biosensor based on imaging ellipsometry , 2009, Journal of viral hepatitis.

[14]  Hans Arwin,et al.  Imaging ellipsometry revisited: Developments for visualization of thin transparent layers on silicon substrates , 1996 .

[15]  Gang Jin,et al.  A label‐free protein microfluidic array for parallel immunoassays , 2006, Electrophoresis.

[16]  Yanyan Chen,et al.  Optimization of off-null ellipsometry for air/solid interfaces. , 2007, Applied optics.

[17]  K. Kirsch,et al.  Genomic organization of the melanoma-associated glycoprotein MUC18: implications for the evolution of the immunoglobulin domains. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[18]  I Lundström,et al.  A biosensor concept based on imaging ellipsometry for visualization of biomolecular interactions. , 1995, Analytical biochemistry.

[19]  Gang Jin,et al.  Detection of avian influenza virus subtype H5 using a biosensor based on imaging ellipsometry. , 2010, Biosensors & bioelectronics.

[20]  Gang Jin,et al.  Phage M13KO7 detection with biosensor based on imaging ellipsometry and AFM microscopic confirmation , 2008, Virus Research.

[21]  Gang Jin,et al.  Feasibility of protein A for the oriented immobilization of immunoglobulin on silicon surface for a biosensor with imaging ellipsometry. , 2003, Journal of biochemical and biophysical methods.

[22]  L. Abrantes,et al.  Immunosensor interface based on physical and chemical immunoglobulin G adsorption onto mixed self-assembled monolayers. , 2006, Bioelectrochemistry.

[23]  D. Elder,et al.  Isolation and functional characterization of the A32 melanoma-associated antigen. , 1994, Cancer research.