Flow imaging microscopy for protein particle analysis--a comparative evaluation of four different analytical...

Area of Science:

• Biopharmaceutical analysis
• Particle characterization
• Microscopy techniques

Background:

• Flow imaging microscopy (FIM) is crucial for protein particle analysis in biopharmaceuticals.
• The market offers several FIM systems, necessitating comparative evaluations for optimal selection.

Purpose of the Study:

• To comparatively evaluate four leading flow imaging microscopy systems: Micro-Flow Imaging (MFI) 4100, MFI5200, Flow Cytometer And Microscope (FlowCAM) VS1, and FlowCAM PV.
• To assess system performance in quantification, characterization, image quality, differentiation of protein particles and silicone oil droplets, and usability.

Main Methods:

• Analysis of polystyrene standards, therapeutic monoclonal antibody particles, and silicone oil droplets using MFI 4100, MFI5200, FlowCAM VS1, and FlowCAM PV.
• Critical performance assessment across key parameters including quantification accuracy, particle differentiation, image resolution, and system handling.

Main Results:

• FlowCAM systems, particularly VS1, provided high-resolution images.
• FlowCAM PV demonstrated superior quantification of monoclonal antibody particles and accurate differentiation of protein particles from silicone oil droplets, even with high refractive index formulations.
• MFI systems, especially MFI5200, offered excellent size and count accuracy with polystyrene standards and robust performance for protein particles.
• MFI systems were noted for ease of use and standardized data analysis compared to FlowCAM systems.

Conclusions:

• The choice of flow imaging microscopy system is application-dependent, based on primary output parameters.
• FlowCAM PV is recommended for precise protein particle quantification and contaminant differentiation.
• MFI systems are suitable for accurate size and count determination and offer a more standardized user experience.