Flow Imaging Microscopy Blog

Characterizing protein aggregates in formulation development

JeffABBIOAB BioTechnologies, Inc. located in Bloomington, Indiana is a privately held laboratory offering pharmaceutical contract services. Founder and CEO Dr. Jeff Schwegman has extensive experience in formulation development, lyophilization (freeze-drying) cycle development and optimization, thermal characterization, and education and training in the development of injectable drug products.

A typical AB BioTechnologies customer needs help with drug and diagnostic products in various stages of development. They can require assistance with a wide variety with problems ranging from, "We just discovered a molecule to treat a disease. What do we do with it?" to, "We’ve got something developed and we're having problems getting it just right."

When AB Biotechnologies completes a project for a customer they hand them a formulation, or a lyophilization cycle and manufacturing process that they can take to a licensed manufacturer to have their product produced.

The Challenge
AB BioTechnologies works with all kinds of customers with a wide variety of backgrounds, but mostly they talk to scientists. They help them when they need answers in regards to development and freeze drying of injectable drugs. They also help business owners doing the initial introduction of a formulation. Sometimes these companies are very small and consist of just a few people. If they don't have scientists on staff they contract AB BioTechnologies to do the bulk of the development work.

"We'll subject formulations to stress factors like freezing and freeze drying and then we'll measure and characterize the initial solution to get a base line," says Dr. Schwegman. "We add stabilizing excipients, then we'll freeze dry it again and reexamine the sample particles to see if we're generating any aggregates."

Protein aggregates can easily go undetected and they have the potential to denature and form aggregates, so finding them is a crucial step in the development process. Dr. Schwegman needs to be able to identify the progression of possible problems in his customer's formulations.

The Solution
AB BioTechnologies uses and recommends laboratory equipment from many vendors, but Dr. Schwegman says that the one most essential to his lab for detecting protein aggregates is the FlowCAM® dynamic imaging particle analysis system from Fluid Imaging Technologies.

When Dr. Schwegman and his team are working on formulations that have the potential to denature and aggregate they do an excipient screen with the FlowCAM. Many protein aggregates are translucent, so this can make them difficult to detect. 

Having the ability to threshold on either darker pixels (relative to the background), lighter pixels or both simultaneously can ensure proper protein aggregate characterization, especially when working with high concentration formulations or those with a large amount of stabilizer.

A traditional method like light obscuration may miss protein aggregates, but the FlowCAM's combination of imaging and flow microscopy allows Dr. Schwegman to characterize particulates with a high level of accuracy. "Image quality is extremely important when characterizing protein aggregates," explained Dr. Schwegman. “We need to be able to differentiate them from silicone oil and other contaminants in drug formulations early in the process.”

The Results
With its VisualSpreadsheet® software, the FlowCAM can record over 30 different measurements per particle and can capture particle images at up to 22 frames per second enabling AB BioTechnologies to quickly get the answers they need to drive the formulation development.

“Data is one thing, but having instantaneous information is another,” says Dr. Schwegman. “The FlowCAM is an integral part of our screening process.”

For example, if they need a do a screening of a monoclonal antibody formulation, they will freeze dry it and see if aggregated particulates show up using the FlowCAM. If aggregated particulates are present, they know they now have to begin blending in some stabilizers. They will repeat the process of blending and running samples to help shape the formulation.

AB Biotechnologies is a relatively small company, but they've been able to grow by finding the right technologies for their lab that help them service more customers more quickly and by offering the best possible solutions. "If the FlowCAM wasn't part of our lab I'd have to contract out if I wanted to test for protein aggregates,” says Dr. Schwegman. “It just made sense to bring it in house."

AB Biotechnologies has experience working with many different formulation types including injectable drug products, lyophilized powders, emulsions, and suspensions. With their development services, a complete, optimized cycle can be designed in less than 30 days. Additionally, Dr. Schwegman provides training in product formulation and lyophilization within the pharmaceutical and diagnostic industries.

AB Biotechnologies is just beginning to learn more about bioanalytical development and is beginning to offer this service to their customers. The next phase for the business is going to be pre-clinical manufacturing, so they are making preparations in their lab to formulate materials for animal studies.

You can email Dr. Schwegman at info@ab-biotech.com or call +1 812-287-8493 or visit AB BioTechnologies at www.ab-biotech.com for information. 

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Topics: Protein Therapeutics, Biopharmaceutical Research