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Biotechnol J. 2017 May 17. doi: 10.1002/biot.201700031. [Epub ahead of print]
Universal label-free In-process Quantification of Influenza Virus-like particles.
Carvalho SB1,2, Moleirinho MG1,2, Wheatley D3, Welsh J3, Gantier R4, Alves PM1,2, Peixoto C1,2, Carrondo MJT1,5.
Author information
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.
Copyright ? 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
KEYWORDS:
Biolayer Interferometry technology; Downstream process; In-process HA quantification; Multivalent VLPs; Octet
PMID: 28514082 DOI: 10.1002/biot.201700031
Universal label-free In-process Quantification of Influenza Virus-like particles.
Carvalho SB1,2, Moleirinho MG1,2, Wheatley D3, Welsh J3, Gantier R4, Alves PM1,2, Peixoto C1,2, Carrondo MJT1,5.
Author information
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.
Copyright ? 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
KEYWORDS:
Biolayer Interferometry technology; Downstream process; In-process HA quantification; Multivalent VLPs; Octet
PMID: 28514082 DOI: 10.1002/biot.201700031