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High-throughput Detection Method for Influenza Virus
Published on: February 4, 2012
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Universal label-free in-process quantification of influenza virus-like particles
Sofia B Carvalho1,2, Mafalda G Moleirinho1,2, David Wheatley3
1iBET, Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal.
Biotechnology Journal
|May 18, 2017
Summary
A new label-free Biolayer interferometry (BLI) method quantifies Influenza virus-like particles (VLPs) efficiently. This high-throughput tool improves bioprocess monitoring and development compared to traditional assays.
Area of Science:
- Biotechnology and Bioprocessing
- Vaccine Development
- Analytical Chemistry
Background:
- Virus-like particles (VLPs) are increasingly used in vaccines, particularly for influenza pandemics, driving demand for advanced manufacturing bioprocesses.
- Current analytical methods for quantifying complex VLPs, such as hemagglutination assay and Single Radial Immunodiffusion, are time-consuming and hinder efficient bioprocess development.
- There is a critical need for rapid, sensitive, and high-throughput analytical tools to support the progress of VLP bioprocessing.
Purpose of the Study:
- To develop and validate a novel, label-free analytical tool for the quantification of Influenza virus-like particles (VLPs) throughout the bioprocess.
- To assess the suitability of Biolayer interferometry (BLI) technology for real-time monitoring and optimization of VLP manufacturing.
- To compare the performance of the BLI method against traditional quantification techniques for Influenza VLPs.
Main Methods:
- Utilized Biolayer interferometry (BLI) on an Octet platform for label-free quantification of Influenza VLPs.
- Employed biotinylated human (α2,6-linked sialic acid) and avian (α2,3-linked sialic acid) receptors immobilized on streptavidin biosensors.
- Quantified hemagglutinin (HA) content in various mono- and multivalent Influenza VLPs from crude samples to final products.
Main Results:
- The BLI method successfully quantified hemagglutinin content in Influenza VLPs across all bioprocess stages.
- Demonstrated high throughput, high sensitivity, and improved detection limits compared to traditional hemagglutination assay and Single Radial Immunodiffusion.
- The method provided real-time results, enabling effective in-line monitoring crucial for downstream processing.
Conclusions:
- Biolayer interferometry (BLI) offers a valuable, rapid, and sensitive label-free tool for quantifying Influenza VLPs.
- This technology significantly enhances downstream process development, control, and optimization by providing real-time analytical data.
- The BLI method represents a substantial advancement over conventional techniques for VLP bioprocess monitoring.
