Finding the Needle in the Haystack: High-Resolution Techniques for Characterization of Mixed Protein Particles...

Area of Science:

• Biopharmaceutical Manufacturing
• Analytical Chemistry
• Materials Science

Background:

• Peristaltic pumps in biopharmaceutical production utilize silicone tubing, which can shed nanometer-sized silicone rubber particles.
• These particles pose a risk of interfering with protein integrity and product quality.
• Previous characterization focused on larger contaminants, leaving nanometer-sized particles undetected.

Purpose of the Study:

• To develop and validate methods for the qualitative and quantitative identification of nanometer-sized silicone particles within protein contaminants.
• To overcome the detection limitations of existing analytical techniques for sub-micrometer contaminants.
• To establish a forensic approach for identifying unknown particle compositions in biopharmaceutical processing.

Main Methods:

• Confocal Raman microscopy for label-free chemical identification and 3D imaging of mixed protein-silicone particles.
• Labeling of silicone tubing to enable high-resolution imaging using confocal laser scanning microscopy.
• Imaging Flow Cytometry for counting mixed protein-silicone particles.

Main Results:

• Demonstrated the capability to detect and identify nanometer-sized silicone particles embedded in or coated by proteins.
• Successfully characterized mixed particles composed of silicone rubber and protein.
• Validated the use of Confocal Raman microscopy and Imaging Flow Cytometry for analyzing these complex contaminants.

Conclusions:

• The developed analytical methods enable the detection and identification of nanometer-sized silicone particles in biopharmaceutical products.
• These techniques provide a valuable forensic tool for troubleshooting contamination issues during biopharmaceutical manufacturing.
• Improved detection of sub-micrometer contaminants enhances biopharmaceutical product safety and quality control.