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Summary

Engineered single domain antibodies (sdAbs) overcome solubility challenges for use in rapid assays. This work optimizes sdAb attachment to nanoparticles and membranes for sensitive biothreat and viral detection.

Keywords:
SARS-CoV-2 nucleocapsidgold nanoparticlericinsingle domain antibodystaphylococcal enterotoxin Bvertical flow assay

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Area of Science:

  • Biotechnology
  • Immunology
  • Assay Development

Background:

  • Single domain antibodies (sdAbs) offer advantages over conventional antibodies, including small size, stability, and cost-effective production.
  • However, sdAbs' high solubility can impede their efficient attachment to surfaces, complicating assay development.
  • Optimizing sdAb integration is crucial for applications like point-of-care testing and field diagnostics.

Purpose of the Study:

  • To engineer single domain antibody (sdAb)-based constructs for improved performance in rapid vertical flow assays.
  • To develop generalizable protocols for attaching sdAbs to gold nanoparticles and support membranes.
  • To enhance the sensitivity and utility of sdAb-based assays for detecting biothreat agents and viral proteins.

Main Methods:

  • Protein engineering of sdAb constructs for enhanced surface adhesion.
  • Development of protocols for conjugating sdAbs to gold nanoparticles.
  • Establishment of methods for immobilizing sdAbs onto support membranes for vertical flow assays.

Main Results:

  • Achieved a limit of detection of 0.11 µg/mL for staphylococcal enterotoxin B and ricin.
  • Successfully detected the nucleocapsid protein of SARS-CoV-2.
  • Demonstrated optimized protocols for sdAb attachment, overcoming previous limitations.

Conclusions:

  • Engineered sdAbs can be effectively integrated into rapid vertical flow assays.
  • The developed protocols facilitate the use of sdAbs in sensitive diagnostic applications.
  • This approach enables robust detection of biothreat agents and viral pathogens in field settings.