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Fluorescence Lifetime Imaging of Molecular Rotors in Living Cells
Published on: February 9, 2012
Antibodies for fluorescent molecular rotors
Biochemistry
|July 27, 1993
Summary
Researchers developed monoclonal antibodies that bind to fluorescent molecular rotors CCVJ and DCVJ. Antibody binding significantly enhances the fluorescence of these molecules by restricting internal rotation, indicating a new method for studying molecular rotors.
Area of Science:
- Biochemistry
- Immunology
- Photophysics
Background:
- Fluorescent molecular rotors like CCVJ and DCVJ are sensitive to their environment.
- Antibodies can be engineered to bind specific small molecules.
Purpose of the Study:
- To prepare monoclonal antibodies against CCVJ and DCVJ.
- To characterize the photophysical properties of CCVJ and DCVJ when bound to antibodies.
Main Methods:
- Production of monoclonal antibodies against CCVJ-conjugated BSA.
- Binding assays using Scatchard plots.
- Fluorescence spectroscopy to measure quantum yield, spectra, and lifetimes.
Main Results:
- Monoclonal antibodies strongly bound CCVJ and DCVJ.
- Antibody binding caused a significant increase in fluorescence quantum yield and lifetime.
- Non-specific IgG did not alter fluorophore properties.
- Association constants were determined for CCVJ binding to IgG and Fab fragments.
Conclusions:
- Antibody binding restricts intramolecular rotation in CCVJ and DCVJ, leading to enhanced fluorescence.
- The observed changes are attributed to reduced non-radiative decay rates.
- This antibody-rotor interaction offers a novel tool for studying molecular rotors.
