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Microbiology JoVE (Open Access) Citable · DOI

Nanomechanics of Drug-target Interactions and Antibacterial Resistance Detection

DOI: 10.3791/50719-v
What you'll learn
  • ✓ Prepare and functionalize cantilever biosensors for drug-target interaction studies
  • ✓ Quantify antibacterial resistance using surface stress detection methodology
  • ✓ Interpret nanomechanical data to assess drug efficacy against resistant bacteria
Protocol

Acquired resistance to antibiotics is a major public healthcare problem and is presently ranked by the WHO as one of the greatest threats to human life. Here we describe the use of cantilever technology to quantify antibacterial resistance, critical to the discovery of novel and powerful agents against multidrug resistant bacteria.

Difficulty
advanced
Total time
~4–6 hours per sample set (cantilever preparation, drug incubation, and measurement cycles)
Biosafety
BSL-2

Steps

1
Prepare and functionalize cantilever biosensors

Clean and chemically functionalize cantilever surfaces to immobilize bacterial antigens or drug targets. This step enables specific binding interactions to be detected.

▶ 02:21
2
Prepare test drug solutions and formulations

Prepare antibiotic and candidate drug formulations at specified concentrations for subsequent binding experiments.

▶ 06:03
3
Detect surface stress and binding events

Flow drug solutions over functionalized cantilevers and measure real-time surface stress changes using optical deflection. Stress variations indicate drug-target binding interactions.

▶ 07:24
4
Quantify drug-target binding interaction data

Analyze cantilever deflection signals to extract binding kinetics and affinity parameters, correlating mechanical changes with antibacterial resistance phenotypes.

▶ 10:05
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