Epitope structure of the carbohydrate recognition domain of asiaglycoprotein receptor to a monoclonal antibody revealed by high resolution proteolytic excision mass spectrometry

Lade...
Vorschaubild
Dateien
Przybylski.pdf
Przybylski.pdfGröße: 4.88 MBDownloads: 1237
Datum
2011
Autor:innen
Stefanescu, Raluca
Born, Rita
Ernst, Beat
Herausgeber:innen
Kontakt
ISSN der Zeitschrift
Electronic ISSN
ISBN
Bibliografische Daten
Verlag
Schriftenreihe
Auflagebezeichnung
ArXiv-ID
Internationale Patentnummer
Angaben zur Forschungsförderung
Projekt
Open Access-Veröffentlichung
Open Access Green
Sammlungen
Core Facility der Universität Konstanz
Gesperrt bis
Titel in einer weiteren Sprache
Forschungsvorhaben
Organisationseinheiten
Zeitschriftenheft
Publikationstyp
Zeitschriftenartikel
Publikationsstatus
Published
Erschienen in
Zusammenfassung

Recent studies suggest that the H1 subunit of the carbohydrate recognition domain (H1CRD) of the asialoglycoprotein receptor is used as entry site into hepatocytes by hepatitis A and B virus, and Marburg virus. Thus, molecules binding specifically to the CRD might exert inhibition towards these diseases by blocking the virus entry site. We report here the identification of the epitope structure of H1CRD to a monoclonal antibody by proteolytic epitope excision of the immune complex and high resolution MALDI-FTICR mass spectrometry. As a prerequisite of the epitope determination, the primary structure of the H1CRD antigen was characterised by ESI-FTICR-MS of the intact protein and by LCMS/MS of tryptic digest mixtures. Molecular mass determination and proteolytic fragments provided the identification of 2 intra-molecular disulfide bridges (7 Cys residues), and a Cysmercaptoethanol adduct formed by treatment with ß-mercaptoethanol during protein extraction. The H1CRD antigen binds to the monoclonal antibody in both native and Cysalkylated form. For identification of the epitope, the antibody was immobilized on N-hydroxysuccinimide activated Sepharose. Epitope- excision and - extraction with trypsin and FTICR-MS of affinity-bound peptides provided the identification of two specific epitope peptides, (5-16) and (17-23) which showed high affinity to the antibody. Affinity studies of the synthetic epitope peptides revealed independent binding of each peptide to the antibody.

Zusammenfassung in einer weiteren Sprache
Fachgebiet (DDC)
540 Chemie
Schlagwörter
Asialoglycoprotein receptor, H1CRD protein, epitope structure, proteolytic epitope excision, high resolution mass spectrometry
Konferenz
Rezension
undefined / . - undefined, undefined
Zitieren
ISO 690STEFANESCU, Raluca, Rita BORN, Adrian MOISE, Beat ERNST, Michael PRZYBYLSKI, 2011. Epitope structure of the carbohydrate recognition domain of asiaglycoprotein receptor to a monoclonal antibody revealed by high resolution proteolytic excision mass spectrometry. In: Journal of The American Society for Mass Spectrometry. 2011, 22(1), pp. 148-157. ISSN 1044-0305. eISSN 1879-1123. Available under: doi: 10.1007/s13361-010-0010-y
BibTex
@article{Stefanescu2011-01Epito-1012,
  year={2011},
  doi={10.1007/s13361-010-0010-y},
  title={Epitope structure of the carbohydrate recognition domain of asiaglycoprotein receptor to a monoclonal antibody revealed by high resolution proteolytic excision mass spectrometry},
  number={1},
  volume={22},
  issn={1044-0305},
  journal={Journal of The American Society for Mass Spectrometry},
  pages={148--157},
  author={Stefanescu, Raluca and Born, Rita and Moise, Adrian and Ernst, Beat and Przybylski, Michael}
}
RDF
<rdf:RDF
    xmlns:dcterms="http://purl.org/dc/terms/"
    xmlns:dc="http://purl.org/dc/elements/1.1/"
    xmlns:rdf="http://www.w3.org/1999/02/22-rdf-syntax-ns#"
    xmlns:bibo="http://purl.org/ontology/bibo/"
    xmlns:dspace="http://digital-repositories.org/ontologies/dspace/0.1.0#"
    xmlns:foaf="http://xmlns.com/foaf/0.1/"
    xmlns:void="http://rdfs.org/ns/void#"
    xmlns:xsd="http://www.w3.org/2001/XMLSchema#" > 
  <rdf:Description rdf:about="https://kops.uni-konstanz.de/server/rdf/resource/123456789/1012">
    <dc:creator>Ernst, Beat</dc:creator>
    <dcterms:abstract xml:lang="eng">Recent studies suggest that the H1 subunit of the carbohydrate recognition domain (H1CRD) of the asialoglycoprotein receptor is used as entry site into hepatocytes by hepatitis A and B virus, and Marburg virus. Thus, molecules binding specifically to the CRD might exert inhibition towards these diseases by blocking the virus entry site. We report here the identification of the epitope structure of H1CRD to a monoclonal antibody by proteolytic epitope excision of the immune complex and high resolution MALDI-FTICR mass spectrometry. As a prerequisite of the epitope determination, the primary structure of the H1CRD antigen was characterised by ESI-FTICR-MS of the intact protein and by LCMS/MS of tryptic digest mixtures. Molecular mass determination and proteolytic fragments provided the identification of 2 intra-molecular disulfide bridges (7 Cys residues), and a Cysmercaptoethanol adduct formed by treatment with ß-mercaptoethanol during protein extraction. The H1CRD antigen binds to the monoclonal antibody in both native and Cysalkylated form. For identification of the epitope, the antibody was immobilized on N-hydroxysuccinimide activated Sepharose. Epitope- excision and - extraction with trypsin and FTICR-MS of affinity-bound peptides provided the identification of two specific epitope peptides, (5-16) and (17-23) which showed high affinity to the antibody. Affinity studies of the synthetic epitope peptides revealed independent binding of each peptide to the antibody.</dcterms:abstract>
    <dc:contributor>Moise, Adrian</dc:contributor>
    <dc:contributor>Przybylski, Michael</dc:contributor>
    <dc:contributor>Born, Rita</dc:contributor>
    <dcterms:available rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-22T17:54:43Z</dcterms:available>
    <dcterms:bibliographicCitation>First publ. in: Journal of the American Society for Mass Spectrometry ; 22 (2011), 1. - pp. 148-157</dcterms:bibliographicCitation>
    <dcterms:hasPart rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/1012/1/Przybylski.pdf"/>
    <dcterms:isPartOf rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dc:contributor>Ernst, Beat</dc:contributor>
    <bibo:uri rdf:resource="http://kops.uni-konstanz.de/handle/123456789/1012"/>
    <dc:creator>Przybylski, Michael</dc:creator>
    <dc:creator>Moise, Adrian</dc:creator>
    <dcterms:rights rdf:resource="https://rightsstatements.org/page/InC/1.0/"/>
    <void:sparqlEndpoint rdf:resource="http://localhost/fuseki/dspace/sparql"/>
    <dc:creator>Stefanescu, Raluca</dc:creator>
    <dc:date rdf:datatype="http://www.w3.org/2001/XMLSchema#dateTime">2011-03-22T17:54:43Z</dc:date>
    <dc:contributor>Stefanescu, Raluca</dc:contributor>
    <dc:creator>Born, Rita</dc:creator>
    <dc:rights>terms-of-use</dc:rights>
    <dcterms:title>Epitope structure of the carbohydrate recognition domain of asiaglycoprotein receptor to a monoclonal antibody revealed by high resolution proteolytic excision mass spectrometry</dcterms:title>
    <foaf:homepage rdf:resource="http://localhost:8080/"/>
    <dcterms:issued>2011-01</dcterms:issued>
    <dspace:isPartOfCollection rdf:resource="https://kops.uni-konstanz.de/server/rdf/resource/123456789/29"/>
    <dspace:hasBitstream rdf:resource="https://kops.uni-konstanz.de/bitstream/123456789/1012/1/Przybylski.pdf"/>
    <dc:language>eng</dc:language>
  </rdf:Description>
</rdf:RDF>
Interner Vermerk
xmlui.Submission.submit.DescribeStep.inputForms.label.kops_note_fromSubmitter
Kontakt
URL der Originalveröffentl.
Prüfdatum der URL
Prüfungsdatum der Dissertation
Finanzierungsart
Kommentar zur Publikation
Allianzlizenz
Corresponding Authors der Uni Konstanz vorhanden
Internationale Co-Autor:innen
Universitätsbibliographie
Ja
Begutachtet
Diese Publikation teilen