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Universal label-free in-process quantification of influenza virus-like particles
Open accessAbstract
Virus‐like particles (VLPs) are becoming established as vaccines, in particular for influenza pandemics, increasing the interest in the development of VLPs manufacturing bioprocess. However, for complex VLPs, the analytical tools used for quantification are not yet able to keep up with the bioprocess progress. Currently, quantification for Influenza relies on traditional methods: hemagglutination assay or Single Radial Immunodiffusion. These analytical technologies are time‐consuming, cumbersome, and not supportive of efficient downstream process development and monitoring. Hereby we report a label‐free tool that uses Biolayer interferometry (BLI) technology applied on an Octet platform to quantify Influenza VLPs at all stages of bioprocess. Human (α2,6‐linked sialic acid) and avian (α2,3‐linked sialic acid) biotinylated receptors associated with streptavidin biosensors were used, to quantify hemagglutinin content in several mono‐ and multivalent Influenza VLPs. The applied method was able to quantify hemagglutinin from crude samples up to final bioprocessing VLP product. BLI technology confirmed its value as a high throughput analytical tool with high sensitivity and improved detection limits compared to traditional methods. This simple and fast method allowed for real‐time results, which are crucial for in‐line monitoring of downstream processing, improving process development, control and optimization.
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BLI is high-throughput, label-free, targeted for at-line use, provides acceptable precision, requires neither sample purification nor preparation, and has been used in process development for biopharmaceuticals and vaccines.
“This high-throughput, label-free method is specifically targeted for at-line use, provides acceptable precision, requires neither sample purification nor preparation, and has been successfully used in process development for biopharmaceuticals and vaccines.”
From Analytical methods for process and product characterization of recombinant adeno-associated virus-based gene therapies (2021) · cites this paper as [108]
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Metadata
- Authors
- Carvalho, S. B., Moleirinho, M. G., Wheatley, D., Welsh, J., Gantier, R., Alves, P. M., Peixoto, C., & Carrondo, M. J. T.
- Affiliations
- Instituto de Biologia Experimental e Tecnológica (PT)
- Pall Corporation (United Kingdom) (GB)
- Pall Life Sciences Westborough MA USA
- Corresponding author
- Peixoto, Cristina
- Venue
- Biotechnology Journal
- Publisher
- Wiley
- Published
- 2017-07-05
- Volume
- 12
- Issue
- 8
- ISSN
- 1860-6768 (ISSN-L)
- 1860-7314
- OpenAlex
- W2615188291
- Cited by
- 30
- References
- 34
- Citations by year
- 2025: 1 · 2024: 3 · 2023: 1 · 2022: 5 · 2021: 5 · 2020: 1 · 2019: 8 · 2018: 4 · 2017: 2
- FWCI
- 3.21
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- Locations
- Biotechnology Journal (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Seventh Framework Programme (EDUFLUVAC project, FP7‐HEALTH‐2013‐INNOVATION)
- Fundação para a Ciência e a Tecnologia (SFRH/BD/52302/2013)
- Topics
- Health Sciences › Medicine › Epidemiology › Influenza Virus Research Studies
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Viral Infectious Diseases and Gene Expression in Insects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Advanced Biosensing Techniques and Applications
- Keywords
- influenza virus-like particles, label-free quantification, real-time process monitoring, bioprocess monitoring, sialic acid receptors, downstream processing, influenza vaccine manufacturing
- MeSH terms
- Antibodies, Viral, Hemagglutinins, Humans, Influenza, Human, Influenza Vaccines, N-Acetylneuraminic Acid, Vaccines, Virus-Like Particle
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