Characterizing Protein Aggregates in Biopharmaceuticals 

Protein Aggregates imaged by FlowCAMParticulates in parenteral drug development have always been a serious issue. In biopharmaceuticals the issue is compounded by reported impacts of aggregates and particles on the product’s efficacy, safety, and immunogenicity.  FDA regulations strongly recommend in-depth characterization of sub-visible particles in protein therapeutics.  New recommendations are specifically targeted at the characterization of sub-visible particles between 2 and 10 microns.

Most protein therapeutics contain a variety of particles, including proteins, silicone drops, and contaminants that have a variety of shapes. Light obscuration, which is typically used to comply with USP <788>, cannot differentiate one particle from another, nor fully characterize particles because the technology assumes all particles are spherical. The FlowCAM imaging particle analysis system allows you to simultaneously determine particle shape, type, and size distribution, plus provides you with an image of each measured.

Image quality is extremely important when characterizing, identifying, and differentiating particles such as protein aggregates, silicone oil, air bubbles, and other contaminants, and it is essential for accurate sizing measurements. Blurry images lead to poor characterization of particles and can affect your particle size distribution. 

The better the overall image quality, the easier it is for the instrument’s software to recognize patterns, and in turn characterize particles.   Blurry images make it difficult to distinguish protein aggregates from silicone droplets and other contaminants, especially in the 2 - 10 µm range.  Poor image quality also makes it difficult for the instrument to discriminate between a single protein amalgamation from several small proteins in close proximity to one another

If particles are not characterized correctly, your size distribution may not be ascertained reliably. Smaller or faint particles (such as those with a low refractive index) may be missed entirely, lowering particle counts.  Large particles may be fractionated into smaller particles - reducing the number of large particles while increasing the number of small particles.

Only the FlowCAM dynamic image particle analysis system, has the image quality necessary to accurately count, size and classify particles in the 2 - 10 µm range.

Download our Image Quality Fact Sheet Now >>

Benefits of the FlowCAM

  • Superior Image Quality - Images from the FlowCAM are the sharpest in the industry, which allows you to distinguish the morphological differences associated with aggregated proteins. You are able to see the difference in morphological properties of protein aggregates, which may indicate bacterial or viral contamination.
  • Flexible and Configurable – Not all proteins look alike. The FlowCAM provides multiple objective / flow cell combinations, so you can optimize the system to suit the formulation under test. Both disposable flow cells and fixed field of view flow cells are available, plus 2X, 4X, 10X, and 20X objectives are also available.
  • High Sensitivity – The FlowCAM provides superior ability to detect particles with a low refractive index because you are able to set threshold limits for dark and light particles. Because protein aggregates can be transparent, there are instances where the aggregate will actually be lighter than the background intensity. FlowCAM allows you to optimize your threshold settings in order to properly capture, measure, and calculate concentration for protein aggregates found in therapeutic proteins, which is especially important when working with high concentration protein samples.

    Learn more about how sensitivity can impact your results in our blog >>

  • Small Sample Volume – The FlowCAM can provide reliable test results with a small sample volume. Our minimum sample volume is only 50 uL, which is especially important when working with precious samples that are in short supply.
  • Superior Software – FlowCAM and Visual Spreadsheet provide industry-leading data analysis capability including the ability to establish custom filters based on over 30 particle properties. Satellite software is also available, so post processing of FlowCAM data can be done at a remote location for added convenience and for sharing data with others.

Learn the 5 important considerations when characterizing protein therapeutics with flow microscopy. Go to article now >>

Article Download

Characterizing Biologics Using Dynamic Imaging Particle Analysis >>
Published in BioPharm International

Compliance done with knowledge and understanding, not by rote: Contract lab solves biopharmaceutical challenges using imaging particle analysis >> 
Published in Processing Magazine

Customer Profiles

Protein Sciences Corporation, Meriden, CT >>
Biotech firm utilizes the FlowCAM because, "the testing process is a whole lot easier, quicker, and more informative, in some ways, than the USP <788> testing.”  The FlowCAM is used as part of their routine testing for sub-visible particles in bulk drug substances and drug products.  When they have a new formulation or process and want to do a stability study, they include the instrument as part of the testing at each point.

AB BioTechnoloiges Inc., Bloomington, IN >>
This privately held laboratory offers pharmaceutical contract services. Their typical customer needs help with drug and diagnostic products in various stages of development.  One of the most essential piece of equipment for detection protein aggregates is the FlowCAM.

Poster Download

Flow Imaging of Subvisible Protein Particles: Moving Towards Best Practices >>
Presented at AAPS Biotech 2013

The Importance of Thresholding in Imaging Analysis of Protein Aggregates >>
Presented at WCBP 2014

A Comparison of Methods for Quantifying Silicone Droplets in Biologics Using Dynamic Imaging Particle Analysis
Presented at the 2014 Workshop on Protein Aggregation and Immunogenicity.

Application Note Download

Detection of Protein Aggregates in Parenteral Drug Formulations >> 

See the FlowCAM in Action!

This quick video will show you how the FlowCAM works and acquires data!