Label-free drug interaction screening via Raman microscopy.

Area of Science:

• Biochemistry
• Spectroscopy
• Drug Discovery

Background:

• Accurate detection of molecular interactions is crucial for drug development.
• Existing methods may require labels or specific conditions, limiting their application.
• A need exists for sensitive, label-free techniques for real-time binding analysis.

Purpose of the Study:

• To develop a simple, label-free screening technique for sensing molecular interactions in solution.
• To establish a method for precise and direct detection of binding across a range of molecule sizes.
• To enable low-concentration and low-dose screening of protein-ligand interactions under physiological conditions.

Main Methods:

• Development of the thermostable Raman interaction profiling (TRIP) technique.
• Application of TRIP to eight protein-ligand systems.
• Analysis of high-resolution Raman measurements using principal component analysis.

Main Results:

• TRIP produced reproducible, high-resolution Raman measurements for protein-ligand binding.
• The technique resolved time-dependent binding between 2,4-dinitrophenol and transthyretin.
• Distinct Raman signals were observed for SARS-CoV-2 spike-antibody interactions, differentiating neutralizing and non-neutralizing antibodies.

Conclusions:

• TRIP is a promising technique for label-free screening of molecular interactions.
• The method facilitates analysis of biologically relevant interactions, including those with viral proteins.
• TRIP holds potential for high-throughput drug screening and real-time binding measurements.