A Novel Synthetic Receptor-Based Immunoassay for Influenza Vaccine Quantification
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{"title"=>"A Novel Synthetic Receptor-Based Immunoassay for Influenza Vaccine Quantification", "type"=>"journal", "authors"=>[{"first_name"=>"Anwar M.", "last_name"=>"Hashem", "scopus_author_id"=>"57200589744"}, {"first_name"=>"Caroline", "last_name"=>"Gravel", "scopus_author_id"=>"23488699900"}, {"first_name"=>"Aaron", "last_name"=>"Farnsworth", "scopus_author_id"=>"35748475200"}, {"first_name"=>"Wei", "last_name"=>"Zou", "scopus_author_id"=>"34772127300"}, {"first_name"=>"Michelle", "last_name"=>"Lemieux", "scopus_author_id"=>"23489278200"}, {"first_name"=>"Kangwei", "last_name"=>"Xu", "scopus_author_id"=>"36646822600"}, {"first_name"=>"Changgui", "last_name"=>"Li", "scopus_author_id"=>"35285745900"}, {"first_name"=>"Junzhi", "last_name"=>"Wang", "scopus_author_id"=>"7701311258"}, {"first_name"=>"Marie France", "last_name"=>"Goneau", "scopus_author_id"=>"55325392600"}, {"first_name"=>"Maria", "last_name"=>"Merziotis", "scopus_author_id"=>"55589413100"}, {"first_name"=>"Runtao", "last_name"=>"He", "scopus_author_id"=>"7202418164"}, {"first_name"=>"Michel", "last_name"=>"Gilbert", "scopus_author_id"=>"7202762328"}, {"first_name"=>"Xuguang", "last_name"=>"Li", "scopus_author_id"=>"35330008400"}], "year"=>2013, "source"=>"PLoS ONE", "identifiers"=>{"isbn"=>"1932-6203", "pmid"=>"23424631", "doi"=>"10.1371/journal.pone.0055428", "pui"=>"368331927", "issn"=>"19326203", "sgr"=>"84873720921", "scopus"=>"2-s2.0-84873720921"}, "id"=>"5ccdc5c3-b3d8-3664-b5da-67d9e1a7b1d1", "abstract"=>"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.", "link"=>"http://www.mendeley.com/research/novel-synthetic-receptorbased-immunoassay-influenza-vaccine-quantification", "reader_count"=>27, "reader_count_by_academic_status"=>{"Librarian"=>1, "Researcher"=>12, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>2, "Student > Master"=>3, "Student > Bachelor"=>3, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_user_role"=>{"Librarian"=>1, "Researcher"=>12, "Student > Ph. D. Student"=>4, "Student > Postgraduate"=>2, "Student > Master"=>3, "Student > Bachelor"=>3, "Lecturer > Senior Lecturer"=>1, "Professor"=>1}, "reader_count_by_subject_area"=>{"Engineering"=>4, "Biochemistry, Genetics and Molecular Biology"=>6, "Materials Science"=>1, "Agricultural and Biological Sciences"=>9, "Medicine and Dentistry"=>4, "Neuroscience"=>1, "Chemistry"=>1, "Immunology and Microbiology"=>1}, "reader_count_by_subdiscipline"=>{"Engineering"=>{"Engineering"=>4}, "Materials Science"=>{"Materials Science"=>1}, "Medicine and Dentistry"=>{"Medicine and Dentistry"=>4}, "Neuroscience"=>{"Neuroscience"=>1}, "Chemistry"=>{"Chemistry"=>1}, "Immunology and Microbiology"=>{"Immunology and Microbiology"=>1}, "Agricultural and Biological Sciences"=>{"Agricultural and Biological Sciences"=>9}, "Biochemistry, Genetics and Molecular Biology"=>{"Biochemistry, Genetics and Molecular Biology"=>6}}, "reader_count_by_country"=>{"Canada"=>1, "France"=>1}, "group_count"=>2}

Scopus | Further Information

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Figshare

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  • {"files"=>["https://ndownloader.figshare.com/files/491011"], "description"=>"<p>Influenza strains included in 2010–2011 vaccine (<b>A</b>) A/California/7/2009(H1N1), (<b>B</b>) A/Victoria/210/2009(H3N2) and (<b>C</b>) B/Brisbane/60/2008(Victoria-like) were incubated at 4°C, 50°C or 100°C for 1 hour, followed by measurements with both SRID and R-ELISA using the corresponding rabbit strain-specific antibodies. Each treatment was tested in triplicate/experiment and the experiment was repeated twice. Results are shown as mean of absorbance from two experiments with error bars indicating the standard deviation. (See also <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055428#pone.0055428.s001\" target=\"_blank\">Figure S1</a>).</p>", "links"=>[], "tags"=>["binding", "synthetic"], "article_id"=>161520, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.g003", "stats"=>{"downloads"=>0, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_heat_treatment_on_vaccine_binding_to_the_synthetic_receptor_/161520", "title"=>"Effect of heat treatment on vaccine binding to the synthetic receptor.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-12 00:25:20"}
  • {"files"=>["https://ndownloader.figshare.com/files/491734"], "description"=>"<p>Each sample was tested in triplicate. Data are shown as mean +/− SD from two experiments.</p><p>HA concentrations were obtained using strain-specific rabbit antibodies.</p>", "links"=>[], "tags"=>["immunology", "Virology", "biotechnology", "physiology"], "article_id"=>162235, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.t003", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Spiking_and_recovery_/162235", "title"=>"Spiking and recovery.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-12 00:37:15"}
  • {"files"=>["https://ndownloader.figshare.com/files/491146"], "description"=>"<p>Influenza strains included in 2010–2011 vaccine (<b>A</b>) A/California/7/2009(H1N1), (<b>B</b>) A/Victoria/210/2009(H3N2) and (<b>C</b>) B/Brisbane/60/2008(Victoria-like) were incubated at the indicated pH for 1 hour and then neutralized to pH 7.2 prior to measurement by both SRID and R-ELISA using the corresponding rabbit strain-specific antibodies. Each treatment was tested in triplicate/experiment and the experiment was repeated at least thrice. Results are shown as mean with error bars indicating the standard deviation.</p>", "links"=>[], "tags"=>["ph", "captured", "synthetic"], "article_id"=>161658, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.g004", "stats"=>{"downloads"=>1, "page_views"=>2, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Effect_of_low_pH_treatment_on_vaccine_captured_by_the_synthetic_receptor_/161658", "title"=>"Effect of low pH treatment on vaccine captured by the synthetic receptor.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-12 00:27:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/491634"], "description"=>"*<p>Monovalent vaccine strain was A/Victoria/210/2009 (H3N2) and trivalent vaccine strain was A/Wisconsin/15/2009 (H3N2).</p>", "links"=>[], "tags"=>["recombinant", "proteins", "antibodies"], "article_id"=>162138, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.t001", "stats"=>{"downloads"=>3, "page_views"=>39, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Vaccines_recombinant_HA_proteins_and_antibodies_used_in_this_study_/162138", "title"=>"Vaccines, recombinant HA proteins and antibodies used in this study.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-12 00:35:38"}
  • {"files"=>["https://ndownloader.figshare.com/files/484712", "https://ndownloader.figshare.com/files/484714", "https://ndownloader.figshare.com/files/484720", "https://ndownloader.figshare.com/files/484728"], "description"=>"<div><p>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.</p> </div>", "links"=>[], "tags"=>["synthetic", "receptor-based", "immunoassay", "influenza", "quantification"], "article_id"=>156547, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>["https://dx.doi.org/10.1371/journal.pone.0055428.s001", "https://dx.doi.org/10.1371/journal.pone.0055428.s002", "https://dx.doi.org/10.1371/journal.pone.0055428.s003", "https://dx.doi.org/10.1371/journal.pone.0055428.s004"], "stats"=>{"downloads"=>0, "page_views"=>42, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/A_Novel_Synthetic_Receptor_Based_Immunoassay_for_Influenza_Vaccine_Quantification__/156547", "title"=>"A Novel Synthetic Receptor-Based Immunoassay for Influenza Vaccine Quantification", "pos_in_sequence"=>0, "defined_type"=>4, "published_date"=>"2013-02-12 01:49:07"}
  • {"files"=>["https://ndownloader.figshare.com/files/491587"], "description"=>"<p>Each sample was tested in triplicate.</p><p>Data are shown as mean (95% Confidence interval).</p>*<p>Monovalent vaccine strain was A/Victoria/210/2009(H3N2) and trivalent vaccine strain was A/Wisconsin/15/2009 (H3N2).</p>¶<p>Expected concentration is labeled values initially determined by SRID.</p>", "links"=>[], "tags"=>["immunology", "Virology", "biotechnology", "physiology"], "article_id"=>162095, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.t005", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Quantification_of_HA_in_vaccines_/162095", "title"=>"Quantification of HA in vaccines.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-12 00:34:55"}
  • {"files"=>["https://ndownloader.figshare.com/files/491267"], "description"=>"<p>Influenza strains included in 2010–2011 vaccine (A/California/7/2009(H1N1), A/Victoria/210/2009(H3N2) and B/Brisbane/60/2008(Victoria-like)) (10 µg/ml) or a trivalent sample (containing 10 µg/ml from each strain) were measured by R-ELISA using (<b>A</b>) Rabbit anti-HA from Influenza A/California/7/2009(H1N1), (<b>B</b>) Rabbit anti-HA from Influenza A/Perth/16/2009 Like(H3N2) or (<b>C</b>) Rabbit anti-HA from Influenza B/Brisbane/60/2008. Each treatment was tested in triplicate/experiment and the experiment was repeated thrice. Results are shown as mean of absorbance with error bars indicating the standard deviation.</p>", "links"=>[], "tags"=>["synthetic", "receptor"], "article_id"=>161776, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.g005", "stats"=>{"downloads"=>1, "page_views"=>3, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Specificity_of_synthetic_receptor_based_ELISA_/161776", "title"=>"Specificity of synthetic receptor based ELISA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-12 00:29:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/490671"], "description"=>"<p>Schematic representation of steps involved in the synthesis and conjugation of the avian and human receptors. A is the avian receptor (α2,3-sialyllactoside-MSA) and B is the human receptor (α2,6-sialyllactoside-MSA). For the ratio of receptors to MSA as estimated by mass spectrometry analysis (MALDI-MS) see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055428#pone-0055428-t002\" target=\"_blank\">Table 2</a>.</p>", "links"=>[], "tags"=>["avian", "influenza", "receptors", "conjugation", "serum", "albumin"], "article_id"=>161176, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.g001", "stats"=>{"downloads"=>0, "page_views"=>1, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Synthesis_of_the_avian_and_human_influenza_receptors_and_conjugation_to_mouse_serum_albumin_MSA_/161176", "title"=>"Synthesis of the avian and human influenza receptors and conjugation to mouse serum albumin (MSA).", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-12 00:19:36"}
  • {"files"=>["https://ndownloader.figshare.com/files/491423"], "description"=>"<p>Three expected ratios (80%, 100% and 120%) of the HA from each influenza reference strain in the 2010–2011 vaccine (<b>A</b>) A/California/7/2009(H1N1), (<b>B</b>) A/Victoria/210/2009(H3N2) and (<b>C</b>) B/Brisbane/60/2008(Victoria-like). Vaccine samples with known amounts of HA (pre-determined by SRID) were employed in spiking and recovery assays using strain-specific rabbit antibodies. Each sample was tested in triplicate. Data are shown as mean ± SD from two experiments (see also <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055428#pone.0055428.s003\" target=\"_blank\">Figure S3</a>). For HA concentrations obtained from each strain at each ratio see <a href=\"http://www.plosone.org/article/info:doi/10.1371/journal.pone.0055428#pone-0055428-t003\" target=\"_blank\">Table 3</a>.</p>", "links"=>[], "tags"=>["synthetic", "receptor"], "article_id"=>161934, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.g006", "stats"=>{"downloads"=>1, "page_views"=>6, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Precision_of_synthetic_receptor_based_ELISA_/161934", "title"=>"Precision of synthetic receptor based ELISA.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-12 00:32:14"}
  • {"files"=>["https://ndownloader.figshare.com/files/491677"], "description"=>"<p>The ratio of receptors to MSA was estimated by mass spectrometry analysis. MSA has a molecular weight of 65,859 Dalton by MALDI-MS, whereas the molecular weights of the α-2,3- and α-2,6-sialyllactoside conjugates are averaged around 79,646 Da and 77,332 Da, respectively, indicating 18 molecules of α-2,3-sialyllactoside are attached to MSA, and 15 of α-2,6-sialyllactoside to MSA.</p>", "links"=>[], "tags"=>["conjugates"], "article_id"=>162186, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.t002", "stats"=>{"downloads"=>1, "page_views"=>9, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Characterization_of_conjugates_by_MALDI_MS_/162186", "title"=>"Characterization of conjugates by MALDI-MS.", "pos_in_sequence"=>0, "defined_type"=>3, "published_date"=>"2013-02-12 00:36:26"}
  • {"files"=>["https://ndownloader.figshare.com/files/490861"], "description"=>"<p>ELISA plates were coated with either (<b>A</b>) the human α2,6-sialyllactoside-MSA receptor or (<b>B</b>) the avian α2,3-sialyllactoside-MSA receptor. Various types of recombinant HA proteins were tested. The binding of each recombinant HA to either receptor was detected with strain-specific antibodies and Uni-1 Abs which are universal antibodies against all HA proteins. Binding is shown as mean of absorbance from three experiments with error bars indicating the standard deviation.</p>", "links"=>[], "tags"=>["specificity", "avian", "synthetic"], "article_id"=>161368, "categories"=>["Physiology", "Biotechnology", "Virology", "Immunology"], "users"=>["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"], "doi"=>"https://dx.doi.org/10.1371/journal.pone.0055428.g002", "stats"=>{"downloads"=>0, "page_views"=>8, "likes"=>0}, "figshare_url"=>"https://figshare.com/articles/_Binding_specificity_of_human_and_avian_synthetic_receptors_/161368", "title"=>"Binding specificity of human and avian synthetic receptors.", "pos_in_sequence"=>0, "defined_type"=>1, "published_date"=>"2013-02-12 00:22:48"}

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Relative Metric

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