Rapid conversion of IgG to biosensor using an antibody-binding protein-based probe.

Area of Science:

• Biochemistry
• Analytical Chemistry
• Immunosensor Technology

Background:

• Antibodies are crucial for specific molecular recognition in immunoassays.
• Developing sensitive and rapid detection methods for small molecules remains a challenge.
• Fluorescent immunosensors offer potential for high-throughput and sensitive detection.

Purpose of the Study:

• To develop a novel fluorescent probe for rapid conversion of antibodies into Quenchbodies.
• To create highly sensitive and specific fluorescent immunosensors for small molecule detection.
• To demonstrate the utility of Quenchbodies for detecting testosterone and ractopamine.

Main Methods:

• Development of a fluorescent probe for antibody conversion.
• Immobilization of converted antibodies onto sensor platforms.
• Antigen-antibody binding assays to determine sensor performance.
• Quantification of testosterone and ractopamine using developed immunosensors.

Main Results:

• Successfully converted full-length antibodies into Quenchbodies, a type of fluorescent immunosensor.
• Developed an anti-testosterone IgG-based immunosensor with a limit of detection (LOD) of 0.76 nM.
• Created an IgG-based immunosensor for ractopamine detection with an LOD of 15.5 pM.
• Both immunosensors exhibited high binding affinity and specificity.

Conclusions:

• The developed fluorescent probe enables rapid generation of Quenchbodies from antibodies.
• Quenchbody-based immunosensors demonstrate high sensitivity and specificity for small molecule detection.
• This technology offers a promising platform for developing novel fluorescent immunosensors.