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Related Experiment Videos

Immobilisation of multilayer bioreceptor assemblies on solid substrates

E Brynda1, M Houska, J Skvor

  • 1Institute of Macromolecular Chemistry, Academy of Sciences of the Czech Republic, Prague, Czech Republic.

Biosensors & Bioelectronics
|May 23, 1998
PubMed
Summary
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Multilayer antibody assemblies enhance horseradish peroxidase detection. Immobilized antibody films show increased binding capacity and reduced non-specific protein adsorption, improving biosensor performance.

Area of Science:

  • Biomaterials Science
  • Immunotechnology
  • Surface Chemistry

Background:

  • Monoclonal antibodies are crucial for specific biomolecule detection.
  • Developing stable and sensitive antibody immobilization techniques is essential for biosensors.
  • Multilayer assembly offers a promising strategy for enhancing antibody-based assays.

Purpose of the Study:

  • To develop and characterize multilayer antibody assemblies for enhanced horseradish peroxidase (HRP) detection.
  • To investigate the binding capacity and stability of immobilized antibodies in multilayer films.
  • To evaluate the non-specific protein adsorption on multilayer antibody surfaces.

Main Methods:

  • Preparation of multilayer assemblies via alternating adsorption of anti-HRP and dextran sulfate (DS) at acid pH.

Related Experiment Videos

  • Crosslinking with glutaraldehyde and selective removal of DS to immobilize antibodies.
  • In situ monitoring of assembly using infrared multi-internal reflection spectroscopy.
  • Quantification of HRP binding capacity using ELISA and optical waveguide light mode spectroscopy.

    • Main Results:

    • The immobilized anti-HRP bilayer exhibited approximately twice the binding activity of a monolayer prepared by simple physiosorption.
    • Further anti-HRP layers increased activity up to 2.5 times that of a monolayer, with diminishing returns.
    • Non-specific adsorption of plasma proteins was three times lower on the multilayer film compared to a physiosorbed monolayer.

    Conclusions:

    • Multilayer assembly of antibodies provides a robust method for enhancing detection sensitivity.
    • The developed immobilization strategy significantly improves antibody-based biosensor performance by increasing specific binding and reducing interference.
    • These findings have implications for developing more effective diagnostic tools and biosensing platforms.