Identification of HIV-1-Based Virus-like Particles by Multifrequency Atomic Force Microscopy

Area of Science:

• Nanotechnology
• Biophysics
• Vaccine Development

Background:

• Virus-like particles (VLPs) are a promising vaccine platform due to their immunogenicity and safety.
• Contamination by cellular vesicles is a significant challenge in VLP production quality control.
• Accurate discrimination between VLPs and cellular vesicles is crucial for reliable vaccine manufacturing.

Purpose of the Study:

• To develop and apply a novel multifrequency atomic force microscopy (MF-AFM) method for VLP characterization.
• To enhance the nanophysical and nanomechanical assessment of HIV-based VLPs.
• To establish a method for differentiating HIV-based VLPs from cellular vesicles for quality control.

Main Methods:

• Utilized multifrequency (MF) atomic force microscopy (AFM) for structural nanophysical characterization of HIV-based VLPs.
• Employed advanced amplitude modulation-frequency modulation (AM-FM) viscoelastic mapping mode for high-resolution imaging.
• Performed particle identification and differentiation under ambient conditions.

Main Results:

• Achieved full structural nanophysical characterization of HIV-based VLPs.
• Enhanced imaging resolution of VLP nanomechanical properties using AM-FM viscoelastic mapping.
• Successfully identified and differentiated HIV-based VLPs from cellular vesicles.

Conclusions:

• MF-AFM provides a novel approach for comprehensive VLP characterization.
• AM-FM viscoelastic mapping offers enhanced insights into VLP biophysical attributes.
• This methodology enables effective quality control and monitoring of VLP preparations, distinguishing them from contaminants.