Covalent immobilization of proteins for the biosensor based on imaging ellipsometry.

In the development of biosensors, the immobilization of biomolecules at interfaces played a crucial role. The feasibility of using 3-aminopropyltriethoxysilane (APTES) and glutaraldehyde (Glu) to modify silicon surface to immobilize covalently protein for immunoassay with the biosensor based on imaging ellipsometry was investigated. The higher density and stability of human IgG layer could be obtained on the silicon surface modified with APTES and Glu than that on the silicon surface modified with dichlorodimethylsilane (DDS). The human IgG molecules immobilized covalently on APTES-Glu surface bound more anti-IgG molecules than that on DDS surface, which indicated that the human IgG molecules could maintain higher binding capability on APTES-Glu surface. Tween 20 was able to block the undesirable adsorption on APTES-Glu surface, and also enhanced the recognition between human IgG and its antibody on both APTES-Glu and DDS surfaces. The combination of this protein covalent immobilization and the biosensor has the potential to be developed into a fast, simple immunoassay technique.

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

[2]  L. Vroman,et al.  Findings with the recording ellipsometer suggesting rapid exchange of specific plasma proteins at liquid/solid interfaces☆ , 1969 .

[3]  J M Calvert,et al.  Use of thiol-terminal silanes and heterobifunctional crosslinkers for immobilization of antibodies on silica surfaces. , 1989, Analytical biochemistry.

[4]  L. Vroman,et al.  Interaction of high molecular weight kininogen, factor XII, and fibrinogen in plasma at interfaces. , 1980, Blood.

[5]  C. Halliwell,et al.  A factorial analysis of silanization conditions for the immobilization of oligonucleotides on glass surfaces. , 2001, Analytical chemistry.

[6]  T. Horbett,et al.  Proteins at Interfaces: An Overview , 1995 .

[7]  S K Bhatia,et al.  Antibody immobilization using heterobifunctional crosslinkers. , 1997, Biosensors & bioelectronics.

[8]  J. Pawliszyn,et al.  Biological sample analysis with immunoaffinity solid-phase microextraction. , 2001, The Analyst.

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

[10]  G. Jin,et al.  [Visualization of the interaction between IL-6 and IL-6R by imaging ellipsometry]. , 2002, Sheng wu gong cheng xue bao = Chinese journal of biotechnology.

[11]  J. Herron,et al.  Characterization of immobilized antibodies on silica surfaces , 1988, IEEE Transactions on Biomedical Engineering.

[12]  A. Doria,et al.  The use of Tween 20 in immunoblotting assays for the detection of autoantibodies in connective tissue diseases. , 2000, Journal of immunological methods.

[13]  H. Funakubo,et al.  Immobilization of ultra-thin layer of monoclonal antibody on glass surface. , 1991, Journal of chromatography.

[14]  Micro-systems for Optical Protein-Chip , 2002 .