Microarray of recombinant antibodies using a streptavidin sensor surface self-assembled onto a gold layer

Area of Science:

• Biotechnology
• Immunology
• Surface Chemistry

Background:

• Recombinant antibody-antigen interactions are crucial in diagnostics and therapeutics.
• Existing detection methods can be time-consuming and require extensive antibody modification.
• Microarray formats offer potential for high-throughput analysis.

Purpose of the Study:

• To develop a sensitive and efficient microarray method for detecting recombinant antibody-antigen interactions.
• To utilize a streptavidin monolayer biochip to minimize nonspecific binding.
• To establish a platform for antibody-antigen kinetics, stability, and screening.

Main Methods:

• Development of a biochip sensor with an oriented streptavidin monolayer.
• Biotinylation of antibody and antigen probes for immobilization on the streptavidin surface.
• Detection of interactions using Cy5-labeled protein targets and kinetic analysis.

Main Results:

• Achieved detection limits of 0.5 µg/mL for peptide antigen and 0.1 µg/mL for recombinant antibodies.
• Demonstrated the ability to measure antibody-antigen kinetics and assess antibody shelf life.
• Established a direct immunoassay for antibody fragments (10-100 nM) and an indirect method for screening.

Conclusions:

• Recombinant antibody fragments are suitable for antibody chip construction.
• The developed microarray method is sensitive, efficient, and reduces the need for antibody purification.
• This platform facilitates large-scale screening of antibody fragments post-expression.