Construction of supported lipid membrane modified piezoelectric biosensor for sensitive assay of cholera toxin based on surface-agglutination of ganglioside-bearing liposomes.

A novel piezoelelctric biosensor has been developed for cholera toxin (CT) detection based on the analyte-mediated surface-agglutination of ganglioside (GM1)-functionalized liposomes. To achieve a CT-specific agglutination at the surface, the gold electrode is modified by a GM1-functionalized supported lipid membrane via spontaneous spread of the liposomes on a self-assembled monolayer of a long-chain alkanethiol. In the presence of CT, the GM1-incorporated liposomes in assay medium will rapidly specifically agglutinate at the electrode surface through the binding of CT to GM1 on the electrode surface and the liposome interface. This results in an enormous mass loading on the piezoelelctric crystal as well as a significant increase of density and viscosity at the interface, thereby generating a decrease in frequency of the piezoelelctric crystal. The combination of mass loading with interfacial change in the surface-agglutination reaction allows the developed piezoelelctric biosensor to show substantial signal amplification in response to the analyte CT. The detection limit can be achieved as low as 25 ng mL(-1) CT. This is the first demonstration on CT detection based on specific surface-agglutination of GM1-modified liposomes. The supported lipid layer based sensing interface can be prepared readily and renewably, making the developed technique especially useful for simple, reusable and sensitive determination of proteins.

[1]  S. Dong,et al.  Sol-gel thin-film immobilized soybean peroxidase biosensor for the amperometric determination of hydrogen peroxide in acid medium. , 1999, Analytical chemistry.

[2]  A. Singh,et al.  Noncompetitive Immunoassays Using Bifunmctional Unilamellar Vesicles or Liposomes , 1995, Biotechnology progress.

[3]  R. Yu,et al.  Immunophenotyping of acute leukemia using an integrated piezoelectric immunosensor array. , 2004, Analytical chemistry.

[4]  Jian-hui Jiang,et al.  A novel piezoelectric immunoagglutination assay technique with antibody-modified liposome. , 2007, Biosensors & bioelectronics.

[5]  Yi Xiao,et al.  Aptamer-functionalized Au nanoparticles for the amplified optical detection of thrombin. , 2004, Journal of the American Chemical Society.

[6]  Sara Tombelli,et al.  Detection of β-thalassemia by a DNA piezoelectric biosensor coupled with polymerase chain reaction , 2003 .

[7]  E. Sackmann,et al.  On the application of supported bilayers as receptive layers for biosensors with electrical detection , 1993 .

[8]  A. Singh,et al.  Application of Antibody and Fluorophore‐Derivatized Liposomes to Heterogeneous Immunoassays for D‐dimer , 1996, Biotechnology progress.

[9]  Jinfang Zhang,et al.  Piezoelectric immunosensor for SARS-associated coronavirus in sputum. , 2004, Analytical chemistry.

[10]  X. Zhou,et al.  Microgravimetric DNA sensor based on quartz crystal microbalance: comparison of oligonucleotide immobilization methods and the application in genetic diagnosis. , 2001, Biosensors & bioelectronics.

[11]  J. Kong,et al.  Ultrathin alumina sol-gel-derived films: allowing direct detection of the liver fibrosis markers by capacitance measurement. , 2003, Analytical chemistry.

[12]  K. Brogden,et al.  Binding between lipopolysaccharide and cecropin A , 1995, Molecular and Cellular Biochemistry.

[13]  G. Shen,et al.  A novel biosensing interfacial design based on the assembled multilayers of the oppositely charged polyelectrolytes , 2005 .

[14]  S. Mukherji,et al.  Immobilization of antibodies on polyaniline films and its application in a piezoelectric immunosensor. , 2006, Analytical chemistry.

[15]  R. Pei,et al.  Assembly of alternating polycation and DNA multilayer films by electrostatic layer-by-layer adsorption. , 2001, Biomacromolecules.

[16]  I. Willner,et al.  Electrochemical and quartz crystal microbalance detection of the cholera toxin employing horseradish peroxidase and GM1-functionalized liposomes. , 2001, Analytical chemistry.

[17]  F. Caruso,et al.  Protein multilayer formation on colloids through a stepwise self-assembly technique. , 1999 .

[18]  Jing-Min Hwang,et al.  Determination of alpha-fetoprotein in human serum by a quartz crystal microbalance-based immunosensor. , 2002, Clinical chemistry.

[19]  Guo-Li Shen,et al.  QCM detection of DNA targets with single-base mutation based on DNA ligase reaction and biocatalyzed deposition amplification. , 2007, Biosensors & bioelectronics.

[20]  Tzong-Zeng Wu,et al.  Artificial receptors in serologic tests for the early diagnosis of dengue virus infection. , 2006, Clinical chemistry.

[21]  D. R. Daniel,et al.  Core-shell nanostructured nanoparticle films as chemically sensitive interfaces. , 2001, Analytical chemistry.

[22]  G McHale,et al.  Molecular-imprinted, polymer-coated quartz crystal microbalances for the detection of terpenes. , 2001, Analytical chemistry.

[23]  Tao Liu,et al.  Particle size effect of the DNA sensor amplified with gold nanoparticles , 2002 .

[24]  I. Willner,et al.  Amplified Microgravimetric Quartz-Crystal-Microbalance Assay of DNA Using Oligonucleotide-Functionalized Liposomes or Biotinylated Liposomes , 2000 .

[25]  Itamar Willner,et al.  Enzyme-Linked Amplified Electrochemical Sensing of Oligonucleotide−DNA Interactions by Means of the Precipitation of an Insoluble Product and Using Impedance Spectroscopy , 1999 .

[26]  C. Pradier,et al.  In-depth investigation of protein adsorption on gold surfaces: correlating the structure and density to the efficiency of the sensing layer. , 2008, The journal of physical chemistry. B.

[27]  R. Yu,et al.  A piezoelectric immunoagglutination assay for Toxoplasma gondii antibodies using gold nanoparticles. , 2004, Biosensors & bioelectronics.

[28]  Huiguang Zhu,et al.  Microscale enzymatic optical biosensors using mass transport limiting nanofilms. 1. Fabrication and characterization using glucose as a model analyte. , 2007, Analytical chemistry.

[29]  F. Caruso,et al.  Quartz crystal microbalance study of DNA immobilization and hybridization for nucleic Acid sensor development. , 1997, Analytical chemistry.

[30]  R. Yu,et al.  Rapid, simple, and sensitive immunoagglutination assay with SiO2 particles and quartz crystal microbalance for quantifying Schistosoma japonicum antibodies. , 2006, Clinical chemistry.

[31]  I. Willner,et al.  Electronic transduction of DNA sensing processes on surfaces: amplification of DNA detection and analysis of single-base mismatches by tagged liposomes. , 2001, Journal of the American Chemical Society.

[32]  Gang Wang,et al.  Application of impedance spectroscopy for monitoring colloid Au-enhanced antibody immobilization and antibody-antigen reactions. , 2004, Biosensors & bioelectronics.

[33]  Y. Fung,et al.  Self-assembled monolayers as the coating in a quartz piezoelectric crystal immunosensor to detect Salmonella in aqueous solution. , 2001, Analytical chemistry.