Universal label-free in-process quantification of influenza virus-like particles

Sofia B. Carvalho, Mafalda G. Moleirinho, David Wheatley, John Welsh, René Gantier, Paula M. Alves, Cristina Peixoto, Manuel J.T. Carrondo

Research output: Contribution to journalArticle

12 Citations (Scopus)

Abstract

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.

Original languageEnglish
Article number1700031
JournalBiotechnology Journal
Volume12
Issue number8
DOIs
Publication statusPublished - 1 Aug 2017

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Orthomyxoviridae
Virion
Interferometry
Hemagglutinins
Technology
Human Influenza
Streptavidin
Immunodiffusion
Hemagglutination
Biosensing Techniques
N-Acetylneuraminic Acid
Pandemics
Limit of Detection
Vaccines

Keywords

  • Biolayer interferometry technology
  • Downstream process
  • In-process HA quantification
  • Multivalent VLPs
  • Octet

Cite this

Carvalho, Sofia B. ; Moleirinho, Mafalda G. ; Wheatley, David ; Welsh, John ; Gantier, René ; Alves, Paula M. ; Peixoto, Cristina ; Carrondo, Manuel J.T. / Universal label-free in-process quantification of influenza virus-like particles. In: Biotechnology Journal. 2017 ; Vol. 12, No. 8.
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Universal label-free in-process quantification of influenza virus-like particles. / Carvalho, Sofia B.; Moleirinho, Mafalda G.; Wheatley, David; Welsh, John; Gantier, René; Alves, Paula M.; Peixoto, Cristina; Carrondo, Manuel J.T.

In: Biotechnology Journal, Vol. 12, No. 8, 1700031, 01.08.2017.

Research output: Contribution to journalArticle

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