Development of an enzyme-linked immunoassay for the quantitation of influenza haemagglutinin: an alternative method to single radial immunodiffusion.

Influenza and Other Respiratory Viruses
Jesse BodleSteven Rockman

Abstract

The current method used to measure haemagglutinin (HA) content for influenza vaccine formulation, single radial immunodiffusion (SRID), is lengthy and relies on the availability of matched standardised homologous reagents. The 2009 influenza pandemic highlighted the need to develop alternate assays that are able to rapidly quantitate HA antigen for vaccine formulation. The aim of this work was to develop an enzyme-linked immunoassay (EIA) for the rapid quantitation of H1, H3, H5 and B influenza HA antigens. Monoclonal antibodies (mAbs) selected for haemagglutination inhibition (HAI) activity were conjugated with horseradish peroxidase and used to establish a capture-detection EIA for the quantitation of HA antigen. Results were compared with the appropriate reference SRID assays to investigate assay performance and utility. Quantitation of HA antigen by EIA correlated well with current reference SRID assays. EIA results showed equivalent precision and exhibited a similar capacity to detect HA antigen in virus samples that had been used in either stability or splitting studies, or subjected to physical or chemical stresses. EIA exhibited greater sensitivity than SRID and has the potential to be used in high-throughput applicatio...Continue Reading

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Citations

Sep 8, 2015·Vaccines·Philip D Minor
Aug 5, 2014·Influenza and Other Respiratory Viruses·Falko SchmeisserJerry P Weir
Mar 17, 2015·Biological Procedures Online·Lianlian Jiang, Maryna C Eichelberger
Jan 1, 2014·Human Vaccines & Immunotherapeutics·Ethan C SettembreRino Rappuoli
Mar 19, 2014·Vaccine·Hyunsuh KimSteven P Rockman
May 18, 2017·Biotechnology Journal·Sofia B CarvalhoManuel J T Carrondo
Jan 23, 2018·Influenza and Other Respiratory Viruses·John M Wood, Jerry P Weir
Dec 11, 2017·Biochemistry. Biokhimii︠a︡·N B UstinovA M Kopylov
Jun 14, 2019·Frontiers in Immunology·Marios KoutsakosKatherine Kedzierska

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