A Rapid and Modular Nanobody Assay for Plug-and-Play Antigen Detection

Area of Science:

• Biotechnology
• Immunodiagnostics
• Molecular Biology

Background:

• Current antigen detection methods like ELISA are sensitive but not portable, while portable lateral flow assays (LFAs) lack adaptability for new targets.
• Traditional diagnostic platforms face challenges in low-resource settings due to complex infrastructure, high costs, and long development times.

Purpose of the Study:

• To develop a versatile, rapid, and portable antigen detection platform overcoming limitations of existing diagnostic tools.
• To create a modular assay adaptable for diverse targets and compatible with on-demand biomanufacturing.

Main Methods:

• Developed the Rapid and Modular Nanobody Assay (RAMONA) using nanobody-coiled coil fusion proteins for modular integration.
• Demonstrated assay generalizability by detecting three distinct protein targets and assessing robustness across temperatures and incubation times.
• Validated compatibility with saliva samples and cell-free protein synthesis systems, enabling multiplexed detection.

Main Results:

• RAMONA achieved detection in under 30 minutes with results strongly correlating to Enzyme-Linked Immunosorbent Assay (ELISA) data.
• The assay demonstrated robustness and compatibility with various sample types and conditions, requiring minimal resources.
• Successful multiplexed detection of three antigens was achieved using orthogonal capture probes.

Conclusions:

• RAMONA offers a significant advancement in rapid, adaptable, and field-deployable antigen detection.
• The platform overcomes limitations of traditional immunoassays, enabling on-demand biomanufacturing and rapid adaptation for diverse targets.
• RAMONA provides a versatile solution for point-of-need diagnostics in various settings.