Strategies for the assessment of protein aggregates in pharmaceutical biotech product development
Open accessAbstract
Within the European Immunogenicity Platform (EIP) ( http://www.e-i-p.eu ), the Protein Characterization Subcommittee (EIP-PCS) has been established to discuss and exchange experience of protein characterization in relation to unwanted immunogenicity. In this commentary, we, as representatives of EIP-PCS, review the current state of methods for analysis of protein aggregates. Moreover, we elaborate on why these methods should be used during product development and make recommendations to the biotech community with regard to strategies for their application during the development of protein therapeutics.
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In a single experiment, AUC assesses presence of aggregates and quantifies relative content of empty, partially filled, and full capsids.
“In a single experiment, AUC assesses the presence of aggregates and quantifies the relative content of empty, partially filled, and full capsids.”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [50]
AUC is generally not quantitative for aggregates below 3%-5%, making analysis of rAAV samples with typical aggregate contents below 1% challenging.
“AUC is generally not quantitative for the quantification of aggregates if their content is below 3%–5%,50 rendering analysis of rAAV samples, with typical aggregate contents below 1%, challenging (Figure 1).”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [50]
SEC-based methods are generally more accurate and precise than orthogonal methods such as AUC or DLS, but accuracy for large aggregates is limited by filtration, nonspecific interactions, and dissociation of reversible aggregates.
“They are generally more accurate and precise than orthogonal methods, e.g., AUC or DLS, but accuracy for large aggregates is limited by the SEC separation mechanism: filtration of aggregates by the stationary phase, nonspecific interactions between aggregates and the column frit, and dissociation of reversible aggregates due to dilution in the mobile phase are common.”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [50]
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- Authors
- den Engelsman, J., Garidel, P., Smulders, R., Koll, H., Smith, B., Bassarab, S., Seidl, A., Hainzl, O., & Jiskoot, W.
- Affiliations
- Boehringer Ingelheim (Germany) (DE)
- MSD (Netherlands) (NL)
- Roche Pharma AG (Germany) (DE)
- UCB Pharma (United Kingdom) (GB)
- Hexal (Germany) (DE)
- Leiden University (NL)
- Corresponding author
- Jiskoot, Wim
- Venue
- Pharmaceutical Research
- Published
- 2010-10-23
- Volume
- 28
- Issue
- 4
- Pages
- 920-933
- ISSN
- 0724-8741 (ISSN-L)
- 1573-904X
- PMC
- PMC3063870
- OpenAlex
- W2167777181
- Cited by
- 348
- References
- 31
- Citations by year
- 2026: 11 · 2025: 10 · 2024: 29 · 2023: 31 · 2022: 19 · 2021: 32 · 2020: 29 · 2019: 20 · 2018: 25 · 2017: 25 · 2016: 19 · 2015: 26 · 2014: 20 · 2013: 19 · 2012: 25
- FWCI
- 6.77
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by-nc
- Locations
- Pharmaceutical Research (publishedVersion, cc-by-nc)
- Europe PMC (PubMed Central) (publishedVersion, cc-by-nc)
- PubMed Central (publishedVersion, cc-by-nc)
- PubMed (publishedVersion)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein purification and stability
- Life Sciences › Immunology and Microbiology › Immunology › Biosimilars and Bioanalytical Methods
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Keywords
- protein aggregates, protein characterization, immunogenicity, therapeutic proteins
- MeSH terms
- Antibodies, Monoclonal, Biotechnology, Drug Contamination, European Union, Immunoassay, Quality Control, Recombinant Proteins, Technology, Pharmaceutical, Guidelines as Topic, Protein Folding, Drug Discovery
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