A novel synthetic receptor-based immunoassay for influenza vaccine quantification.
Vaccination is the most effective prophylactic method for preventing influenza. Quantification of influenza vaccine antigens is critically important before the vaccine is used for human immunization. Currently the vaccine antigen quantification relies on hemagglutinin content quantification, the key...
| Main Authors: | , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
Public Library of Science (PLoS)
2013-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC3570553?pdf=render |
| _version_ | 1819040438794846208 |
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| author | Anwar M Hashem Caroline Gravel Aaron Farnsworth Wei Zou Michelle Lemieux Kangwei Xu Changgui Li Junzhi Wang Marie-France Goneau Maria Merziotis Runtao He Michel Gilbert Xuguang Li |
| author_facet | Anwar M Hashem Caroline Gravel Aaron Farnsworth Wei Zou Michelle Lemieux Kangwei Xu Changgui Li Junzhi Wang Marie-France Goneau Maria Merziotis Runtao He Michel Gilbert Xuguang Li |
| author_sort | Anwar M Hashem |
| collection | DOAJ |
| description | Vaccination is the most effective prophylactic method for preventing influenza. Quantification of influenza vaccine antigens is critically important before the vaccine is used for human immunization. Currently the vaccine antigen quantification relies on hemagglutinin content quantification, the key antigenic component, by single radial immunodiffusion (SRID) assay. Due to the inherent disadvantages associated with the traditional SRID; i.e. low sensitivity, low throughput and need for annual reagents, several approaches have been proposed and investigated as alternatives. Yet, most alternative methods cannot distinguish native hemagglutinin from denatured form, making them less relevant to antigenic analyses. Here, we developed a quantitative immunoassay based on the sialic acid binding property of influenza vaccine antigens. Specifically, we chemically synthesized human and avian influenza virus receptors analogues, N-acetylneuraminic acid-2,6-lactose and N-acetylneuraminic acid-2,3-lactose derivatives with an azidopropyl aglycon, using α-2,6- and α-2,3-sialyltransferases, respectively. The azido group of the two sialyllactose-derivatives was reduced and conjugated to mouse serum albumin through a squarate linkage. We showed that the synthetic α-2,6- and α-2,3-receptors selectively bound to human and avian-derived hemagglutinins, respectively, forming the basis of a new, and robust assay for hemagglutinin quantification. Hemagglutinin treated at high temperature or low pH was measured differentially to untreated samples suggesting native conformation is dependent for optimal binding. Importantly, this receptor-based immunoassay showed excellent specificity and reproducibility, high precision, less turnaround time and significantly higher sensitivity and throughput compared with SRID in analyzing multiple influenza vaccines. |
| first_indexed | 2024-12-21T09:09:06Z |
| format | Article |
| id | doaj.art-e331e891d2c84da28b2d64a826364428 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-21T09:09:06Z |
| publishDate | 2013-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-e331e891d2c84da28b2d64a8263644282022-12-21T19:09:17ZengPublic Library of Science (PLoS)PLoS ONE1932-62032013-01-0182e5542810.1371/journal.pone.0055428A novel synthetic receptor-based immunoassay for influenza vaccine quantification.Anwar M HashemCaroline GravelAaron FarnsworthWei ZouMichelle LemieuxKangwei XuChanggui LiJunzhi WangMarie-France GoneauMaria MerziotisRuntao HeMichel GilbertXuguang LiVaccination is the most effective prophylactic method for preventing influenza. Quantification of influenza vaccine antigens is critically important before the vaccine is used for human immunization. Currently the vaccine antigen quantification relies on hemagglutinin content quantification, the key antigenic component, by single radial immunodiffusion (SRID) assay. Due to the inherent disadvantages associated with the traditional SRID; i.e. low sensitivity, low throughput and need for annual reagents, several approaches have been proposed and investigated as alternatives. Yet, most alternative methods cannot distinguish native hemagglutinin from denatured form, making them less relevant to antigenic analyses. Here, we developed a quantitative immunoassay based on the sialic acid binding property of influenza vaccine antigens. Specifically, we chemically synthesized human and avian influenza virus receptors analogues, N-acetylneuraminic acid-2,6-lactose and N-acetylneuraminic acid-2,3-lactose derivatives with an azidopropyl aglycon, using α-2,6- and α-2,3-sialyltransferases, respectively. The azido group of the two sialyllactose-derivatives was reduced and conjugated to mouse serum albumin through a squarate linkage. We showed that the synthetic α-2,6- and α-2,3-receptors selectively bound to human and avian-derived hemagglutinins, respectively, forming the basis of a new, and robust assay for hemagglutinin quantification. Hemagglutinin treated at high temperature or low pH was measured differentially to untreated samples suggesting native conformation is dependent for optimal binding. Importantly, this receptor-based immunoassay showed excellent specificity and reproducibility, high precision, less turnaround time and significantly higher sensitivity and throughput compared with SRID in analyzing multiple influenza vaccines.http://europepmc.org/articles/PMC3570553?pdf=render |
| spellingShingle | Anwar M Hashem Caroline Gravel Aaron Farnsworth Wei Zou Michelle Lemieux Kangwei Xu Changgui Li Junzhi Wang Marie-France Goneau Maria Merziotis Runtao He Michel Gilbert Xuguang Li A novel synthetic receptor-based immunoassay for influenza vaccine quantification. PLoS ONE |
| title | A novel synthetic receptor-based immunoassay for influenza vaccine quantification. |
| title_full | A novel synthetic receptor-based immunoassay for influenza vaccine quantification. |
| title_fullStr | A novel synthetic receptor-based immunoassay for influenza vaccine quantification. |
| title_full_unstemmed | A novel synthetic receptor-based immunoassay for influenza vaccine quantification. |
| title_short | A novel synthetic receptor-based immunoassay for influenza vaccine quantification. |
| title_sort | novel synthetic receptor based immunoassay for influenza vaccine quantification |
| url | http://europepmc.org/articles/PMC3570553?pdf=render |
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