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Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin

BACKGROUND: Alternative methods are being sought to measure the potency of influenza vaccines. Label-free technologies that do not require the use of hemagglutinin (HA)-specific antisera are particularly attractive as the preparation of antiserum delays availability of potency reagents. The objectiv...

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Autores principales: Jiang, Lianlian, Eichelberger, Maryna C
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4359790/
https://www.ncbi.nlm.nih.gov/pubmed/25774096
http://dx.doi.org/10.1186/s12575-015-0019-5
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author Jiang, Lianlian
Eichelberger, Maryna C
author_facet Jiang, Lianlian
Eichelberger, Maryna C
author_sort Jiang, Lianlian
collection PubMed
description BACKGROUND: Alternative methods are being sought to measure the potency of influenza vaccines. Label-free technologies that do not require the use of hemagglutinin (HA)-specific antisera are particularly attractive as the preparation of antiserum delays availability of potency reagents. The objective of these experiments was to evaluate the use of a Corning Epic® label-free method to quantify functional influenza hemagglutinin in rHA preparations. The method was optimized to quantify recombinant HA (rHA) of B/Brisbane/60/2008 (B/BR/08). Fetuin was immobilized onto plates and the change in wavelength of refracted light measured using an Enspire (Perkin Elmer) instrument. RESULTS: The change in wavelength measured in response to addition of rHA of B/BR/08 was proportional to its concentration and was optimal in the presence of native rHA conformations. However, the assay was strain-dependent and did not correlate with HAU measured using turkey red blood cells. CONCLUSIONS: The Corning Epic® label-free method is suitable for quantifying the native forms of rHA for B/BR/08 and A/Brisbane/59/2007 (H1N1) and A/Hangxhou/3/2013 (H7N9). This method is a useful tool for research purposes but further investigation is needed to identify suitable glycoproteins to use as ligands that allow quantification of HAs from a broader range of virus strains. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12575-015-0019-5) contains supplementary material, which is available to authorized users.
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spelling pubmed-43597902015-03-16 Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin Jiang, Lianlian Eichelberger, Maryna C Biol Proced Online Research BACKGROUND: Alternative methods are being sought to measure the potency of influenza vaccines. Label-free technologies that do not require the use of hemagglutinin (HA)-specific antisera are particularly attractive as the preparation of antiserum delays availability of potency reagents. The objective of these experiments was to evaluate the use of a Corning Epic® label-free method to quantify functional influenza hemagglutinin in rHA preparations. The method was optimized to quantify recombinant HA (rHA) of B/Brisbane/60/2008 (B/BR/08). Fetuin was immobilized onto plates and the change in wavelength of refracted light measured using an Enspire (Perkin Elmer) instrument. RESULTS: The change in wavelength measured in response to addition of rHA of B/BR/08 was proportional to its concentration and was optimal in the presence of native rHA conformations. However, the assay was strain-dependent and did not correlate with HAU measured using turkey red blood cells. CONCLUSIONS: The Corning Epic® label-free method is suitable for quantifying the native forms of rHA for B/BR/08 and A/Brisbane/59/2007 (H1N1) and A/Hangxhou/3/2013 (H7N9). This method is a useful tool for research purposes but further investigation is needed to identify suitable glycoproteins to use as ligands that allow quantification of HAs from a broader range of virus strains. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12575-015-0019-5) contains supplementary material, which is available to authorized users. BioMed Central 2015-03-09 /pmc/articles/PMC4359790/ /pubmed/25774096 http://dx.doi.org/10.1186/s12575-015-0019-5 Text en © Jiang and Eichelberger; licensee BioMed Central. 2015 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Jiang, Lianlian
Eichelberger, Maryna C
Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin
title Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin
title_full Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin
title_fullStr Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin
title_full_unstemmed Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin
title_short Evaluation of Epic® label-free technology to quantify functional recombinant hemagglutinin
title_sort evaluation of epic® label-free technology to quantify functional recombinant hemagglutinin
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4359790/
https://www.ncbi.nlm.nih.gov/pubmed/25774096
http://dx.doi.org/10.1186/s12575-015-0019-5
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