Buffer system improves the removal of host cell protein impurities in monoclonal antibody purification

Buffer system improves the removal of host cell protein impurities in monoclonal antibody purification

Polysorbates (PS) are commonly used as stabilizers of biopharmaceuticals such as monoclonal antibodies (mAbs). However, they are prone to chemical and enzymatic degradation. The latter can be caused by residual host cell proteins (HCPs) in the drug substance. Degradation affects the functionality of...

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Bibliographic Details
Published in:Biotechnology and bioengineering Vol. 121; no. 12; pp. 3869 - 3880
Main Authors: Lakatos, Dániel, Idler, Martina, Stibitzky, Selina, Amann, Jennifer, Schuschkewitz, Jakob, Krayl, Dominik, Liebau, Judith, Grosch, Jan-Hendrik, Arango Gutierrez, Erik, Kluters, Simon
Format: Journal Article
Language:English
Published: United States Wiley Subscription Services, Inc 01-12-2024
Subjects:
Acetic acid
Affinity
Affinity chromatography
Anion exchanging
Biopharmaceuticals
Chromatography
Degradation
DNA viruses
Elution
Glycine
Impurities
Inactivation
Industrial development
Monoclonal antibodies
Precipitates
Protein A
Protein purification
Proteins
Turbidity
polysorbate degradation
acetate buffer
glycine buffer
Host Cell Proteins (HCP)
Monoclonal Antibody (mAb)
Protein A affinity chromatography
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Summary:Polysorbates (PS) are commonly used as stabilizers of biopharmaceuticals such as monoclonal antibodies (mAbs). However, they are prone to chemical and enzymatic degradation. The latter can be caused by residual host cell proteins (HCPs) in the drug substance. Degradation affects the functionality of the PS surfactant which can lead to formation of particles. An increasing number of publications describe enzymatic PS degradation. Significant efforts have been made to characterize HCP removal during Downstream Processing (DSP) of mAbs and to develop mitigation strategies. Here we describe the use of glycine buffer for acidic elution in Protein A affinity chromatography compared to acetate buffer, which is more commonly used in the biopharmaceutical industry. Increased turbidity was observed during pH re-adjustment after low pH virus inactivation when using glycine buffer. Analytical data suggests that this turbidity is caused by the formation of precipitates which include HCP and DNA impurities. Additionally, as a zwitterion, glycine does not contribute to conductivity; this further enhances HCP removal during anion-exchange flow-through chromatography. Although glycine is well known as a possible elution buffer for Protein A affinity chromatography, its positive impact on HCP removal and PS stability have not yet been described in literature.
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ISSN:0006-3592
1097-0290
1097-0290
DOI:10.1002/bit.28844