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Nonlinear immunofluorescent assay for androgenic hormones based on resonant structures
Anisha Thayil1, Alejandro Muriano, J-Pablo Salvador
1ICFO-Institut de Ciencies Fotoniques, Ada Canal Olimpic s/n, 08600 Castelldefels, Barcelona, Spain.
Optics Express
|August 20, 2008
Summary
This study introduces two-photon fluorescence detection for affinity binding reactions. A resonant grating waveguide platform successfully detected fluorescent-labeled Boldenone interacting with antibodies, achieving high sensitivity.
Area of Science:
- Biophysics
- Analytical Chemistry
- Biotechnology
Background:
- Affinity binding reactions are crucial in diagnostics and drug discovery.
- Sensitive detection methods are needed for analyzing biomolecular interactions.
- Non-natural hormones like Boldenone require specific detection strategies.
Purpose of the Study:
- To demonstrate two-photon fluorescence as a novel detection method for affinity binding.
- To utilize a resonant grating waveguide structure for enhanced signal detection.
- To detect the interaction between fluorescent-labeled Boldenone and its specific antibody.
Main Methods:
- Employing two-photon fluorescence microscopy for detection.
- Utilizing a resonant grating waveguide structure for signal amplification.
- Immobilizing anti-anabolic antibodies on the waveguide surface.
- Incubating with fluorescently labeled Boldenone to detect binding.
Main Results:
- Successfully detected affinity binding reactions using two-photon fluorescence.
- Achieved a surface coverage detection limit of approximately 0.7 ng/mm(2).
- Demonstrated the specificity of the antibody-hormone interaction.
Conclusions:
- Two-photon fluorescence is a viable and sensitive detection method for affinity binding.
- Resonant grating waveguides enhance detection sensitivity for biomolecular interactions.
- This method offers potential for detecting non-natural hormones and related biomarkers.
