A Polishing Strategy for Removing Impurities in Bispecific Antibody Purification

Bispecific antibodies (bsAbs) are therapeutically promising due to their ability to bind to two different antigens. Although bsAb molecules provide huge therapeutic advantages over traditional monoclonal antibodies (MAbs), their complex product-and-process-related impurities pose unique challenges to their downstream processing.
Here, using two knob-into-hole (KiH) based asymmetric IgG-like bsAb and one symmetric IgG-like bsAb constructs as model molecules, we demonstrated an effective bsAb polishing strategy leveraging on multimodal CHT™ Ceramic Hydroxyapatite chromatography media. Under optimized conditions, excellent removal of bsAb byproducts and impurities were achieved in a single chromatographic step, with eluate purity in all products over 97% and HMW (high molecular weight) species less than 0.6%. In addition, the chromatography step yielded purified products with low HCP contents (below 100 ppm), and HCDNA levels for all products post-CHT resins were lower than 10 ppb. The study illustrates that CHT chromatography can be utilized as an effective polishing step for not only MAbs, but also bsAbs, covering both asymmetric and symmetric formats.
The results presented here would be an insightful guidance to all researchers working on the purification process development to produce bsAbs, especially for asymmetric and symmetric IgG-like bsAb therapeutics.
Key Takeaways:
- Three post Protein A bsAbs were polished by one-step CHT to over 97% purity.
- Provided efficient removal of both product-and-process-related impurities.
- Minimized “chromatography-induced aggregation” for aggregation-prone bsAbs.
Learn more at www.bio-rad.com/library
We Are Bio-Rad.
Our mission: To provide useful, high-quality products and services that advance scientific discovery and improve healthcare. At Bio-Rad, we are united behind this effort. These two objectives are the driving force behind every decision we make, from developing innovative ideas to building global solutions that help solve our customers' greatest challenges.
Connect with Bio-Rad Online:
Website: www.bio-rad.com/
LinkedIn: / 1613226
Facebook: / biorad
Twitter: / bioradlifesci
Instagram: @BioRadLabs
Snapchat: @BioRadLabs
©2024 Bio-Rad is a trademark of Bio-Rad Laboratories, Inc. in certain jurisdictions. All trademarks used herein are the property of their respective owner.
Music used by permission granted by royalty-free license from Pond5.com