A Novel Synthetic Receptor-Based Immunoassay for Influenza Vaccine Quantification
| Autor: | Michelle Lemieux, Michel Gilbert, Kangwei Xu, Xuguang Li, Changgui Li, Wei Zou, Junzhi Wang, Runtao He, Maria Merziotis, Marie-France Goneau, Caroline Gravel, Anwar M. Hashem, Aaron Farnsworth |
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| Jazyk: | angličtina |
| Rok vydání: | 2013 |
| Předmět: |
Protein Denaturation
Anatomy and Physiology lcsh:Medicine Hemagglutinin Glycoproteins Influenza Virus avian influenza virus medicine.disease_cause chemistry.chemical_compound Influenza A Virus H1N1 Subtype sensitivity analysis Immune Physiology Influenza A virus Glycosides lcsh:Science Antigens Viral beta-D-Galactoside alpha 2-6-Sialyltransferase Radial immunodiffusion alpha 2 6 sialyltransferase Vaccines Multidisciplinary medicine.diagnostic_test quantitative analysis Vaccination n acetylneuraminic acid 2 3 lactose influenza vaccination sialic acid Influenza Vaccines protein protein interaction Medicine alpha 2 6 sialyllactoside receptor influenza temperature measurement Research Article Biotechnology Azides Immunodiffusion beta-Galactoside alpha-2 3-Sialyltransferase Influenza vaccine serum albumin Immunology Hemagglutinin (influenza) Enzyme-Linked Immunosorbent Assay Sialic acid binding Biology Microbiology virus receptor Birds sialylation Species Specificity Virology medicine alpha 2 3 sialyllactoside receptor Animals Humans linkage analysis n acetylneuraminic acid 2 6 lactose Antigens Immunoassays Protein Structure Quaternary virus hemagglutinin alpha 2 3 sialyltransferase Influenza A Virus H3N2 Subtype lcsh:R pH measurement Immunity nucleotide sequence Viral Vaccines n acetylneuraminic acid derivative N-Acetylneuraminic Acid Sialyltransferases enzyme linked immunosorbent assay chemistry sensitivity and specificity Immunoassay Influenza in Birds biology.protein Immunologic Techniques antigen specificity lcsh:Q Clinical Immunology Protein Multimerization N-Acetylneuraminic acid |
| Zdroj: | PLoS ONE PLoS ONE, Vol 8, Iss 2, p e55428 (2013) |
| ISSN: | 1932-6203 |
| Popis: | 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. © 2013 Hashem et al. |
| Databáze: | OpenAIRE |
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