Switchable reporter enzymes based on mutually exclusive domain interactions allow antibody detection directly in...

Area of Science:

• Biochemistry
• Immunology
• Enzyme Engineering

Background:

• Antibody detection is crucial for diagnosing infections, allergies, and autoimmune diseases.
• Existing immunoassays are complex, involving multiple steps and limiting point-of-care applications.

Purpose of the Study:

• To develop a simplified method for antibody detection using switchable reporter enzymes.
• To enable direct, colorimetric detection of antibodies in solution, improving diagnostic speed and accessibility.

Main Methods:

• Engineered switchable reporter enzymes based on TEM1 β-lactamase and its inhibitor protein BLIP.
• Utilized antibody-induced disruption of the enzyme-inhibitor interaction for signal generation.
• Demonstrated modularity by creating reporters for HIV1-p17, HA-tag, and Dengue virus type I antibodies.

Main Results:

• Achieved a 10-fold increase in reporter enzyme activity at picomolar antibody concentrations.
• Successfully generated reporter enzymes for multiple antibody targets through epitope sequence replacement.
• Validated a generic and modular approach for antibody reporter enzyme construction.

Conclusions:

• The developed switchable reporter enzymes offer a simplified, generic, and modular platform for antibody detection.
• This approach eliminates cumbersome optimization steps, facilitating broader applications in diagnostics and screening.
• Enables direct colorimetric detection, advancing point-of-care and high-throughput diagnostic capabilities.