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Enzyme-Linked Immunosorbent Assay

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Determination of High-affinity Antibody-antigen Binding Kinetics Using Four Biosensor Platforms

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Published on: April 17, 2017

Light assisted antibody immobilization for bio-sensing.

Bartolomeo Della Ventura, Luigi Schiavo, Carlo Altucci

Biomedical Optics Express

|November 15, 2011

Summary

Ultrashort UV pulses selectively break disulfide bonds in antibodies, enabling efficient surface anchoring for biosensors. This method enhances Quartz Crystal Microbalance (QCM) sensor sensitivity and linear range.

Keywords:

(170.0170) Medical optics and biotechnology (280.1415) Biological sensing and sensors (350.3450) Laser-induced chemistry

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Analyzing DNA-Protein Interactions with Streptavidin-Based Biolayer Interferometry

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Published on: January 17, 2025

Area of Science:

Biochemistry
Materials Science
Analytical Chemistry

Background:

Antibody immobilization is crucial for biosensor development.
Disulfide bonds play a role in protein structure and reactivity.
Current methods for surface anchoring can be inefficient.

Purpose of the Study:

To investigate the use of ultrashort UV pulses for antibody immobilization.
To enhance the performance of Quartz Crystal Microbalance (QCM) sensors.
To explore novel surface functionalization techniques.

Main Methods:

Irradiation of antibody solutions with 258 nm ultrashort UV pulses.
Anchoring of UV-treated antibodies to gold-plated electrodes.
Analysis of disulfide bond breakage using chemical assays.
Performance evaluation of antibody-functionalized QCM sensors.

Main Results:

UV pulses effectively disrupted disulfide bridges in antibodies.
Disrupted disulfide bonds facilitated efficient antibody anchoring to gold surfaces.
UV-irradiated antibodies showed preferential orientation on the electrode.
QCM sensors functionalized with UV-treated antibodies exhibited increased sensitivity and linear range.

Conclusions:

Ultrashort UV pulse irradiation offers a controlled method for antibody immobilization via disulfide bond disruption.
This technique enhances biosensor performance, particularly for QCM-based devices.
The combination of cysteine and tryptophan residues provides a versatile "hook" for immobilizing challenging biomolecules.