Production of serotype C specific and serotype C/D generic monoclonalantibodies using recombinant HC and HN fragments from Clostridiumbotulinum neurotoxin types C1 and D

Rhonda M Curran, E Fringuelli, David Graham, Chris T Elliott

    Research output: Contribution to journalArticle

    4 Citations (Scopus)

    Abstract

    Sequence variability of Clostridium botulinum serotypes C and D is particularly complex.Some serotype C and D strains have unique gene structures that encode mosaic isoforms ofbotulinum neurotoxin (BoNT) containing components of both BoNT type C1 (BoNT/C1) andBoNT type D (BoNT/D). Such sequence variability and the potential for cross neutralisationmust be taken into consideration when developing serotype C and D detection andidentification assays. Three fusion proteins containing either a fragment from thecarboxyl-terminal domain of the heavy chain (HC) of BoNT/C1 (strain 573), a fragment fromthe HC of BoNT/D (strain BVD/-3) or a fragment from the amino-terminal domain of theheavy chain (HN) of BoNT/C1 (strain 573) were expressed in Escherichia coli, andadministered as immunogens to mice. Monoclonal antibodies (mAbs) against therecombinant BoNT fragments were prepared by three fusions. MAbs recognising nativeBoNT/C1 and BoNT/D were detected by enzyme-linked immunosorbent assay (ELISA). Ninemonoclonal antibodies (mAbs) were produced, six of which recognised a BoNT fragmentthat is highly conserved across all serotype C and D producing strains. We conclude thatthese mAbs and this approach to mAb production may facilitate the development ofimmunological diagnostic techniques that are not constrained by the existence of mosaicisoforms for the detection and identification of serotypes C and D.
    LanguageEnglish
    Pages1-10
    JournalVeterinary Immunology and Immunopathology
    Volume130
    DOIs
    Publication statusPublished - 2009

    Fingerprint

    Monoclonal Antibodies
    Clostridium botulinum type D
    Clostridium botulinum type C
    Neurotoxins
    Protein Isoforms
    Enzyme-Linked Immunosorbent Assay
    Serogroup
    cobra toxin D
    Escherichia coli
    Antibodies
    Genes
    Proteins

    Cite this

    @article{7764f18e72384bd6bc5c470442f23841,
    title = "Production of serotype C specific and serotype C/D generic monoclonalantibodies using recombinant HC and HN fragments from Clostridiumbotulinum neurotoxin types C1 and D",
    abstract = "Sequence variability of Clostridium botulinum serotypes C and D is particularly complex.Some serotype C and D strains have unique gene structures that encode mosaic isoforms ofbotulinum neurotoxin (BoNT) containing components of both BoNT type C1 (BoNT/C1) andBoNT type D (BoNT/D). Such sequence variability and the potential for cross neutralisationmust be taken into consideration when developing serotype C and D detection andidentification assays. Three fusion proteins containing either a fragment from thecarboxyl-terminal domain of the heavy chain (HC) of BoNT/C1 (strain 573), a fragment fromthe HC of BoNT/D (strain BVD/-3) or a fragment from the amino-terminal domain of theheavy chain (HN) of BoNT/C1 (strain 573) were expressed in Escherichia coli, andadministered as immunogens to mice. Monoclonal antibodies (mAbs) against therecombinant BoNT fragments were prepared by three fusions. MAbs recognising nativeBoNT/C1 and BoNT/D were detected by enzyme-linked immunosorbent assay (ELISA). Ninemonoclonal antibodies (mAbs) were produced, six of which recognised a BoNT fragmentthat is highly conserved across all serotype C and D producing strains. We conclude thatthese mAbs and this approach to mAb production may facilitate the development ofimmunological diagnostic techniques that are not constrained by the existence of mosaicisoforms for the detection and identification of serotypes C and D.",
    author = "Curran, {Rhonda M} and E Fringuelli and David Graham and Elliott, {Chris T}",
    year = "2009",
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    Production of serotype C specific and serotype C/D generic monoclonalantibodies using recombinant HC and HN fragments from Clostridiumbotulinum neurotoxin types C1 and D. / Curran, Rhonda M; Fringuelli, E; Graham, David; Elliott, Chris T.

    In: Veterinary Immunology and Immunopathology, Vol. 130, 2009, p. 1-10.

    Research output: Contribution to journalArticle

    TY - JOUR

    T1 - Production of serotype C specific and serotype C/D generic monoclonalantibodies using recombinant HC and HN fragments from Clostridiumbotulinum neurotoxin types C1 and D

    AU - Curran, Rhonda M

    AU - Fringuelli, E

    AU - Graham, David

    AU - Elliott, Chris T

    PY - 2009

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    AB - Sequence variability of Clostridium botulinum serotypes C and D is particularly complex.Some serotype C and D strains have unique gene structures that encode mosaic isoforms ofbotulinum neurotoxin (BoNT) containing components of both BoNT type C1 (BoNT/C1) andBoNT type D (BoNT/D). Such sequence variability and the potential for cross neutralisationmust be taken into consideration when developing serotype C and D detection andidentification assays. Three fusion proteins containing either a fragment from thecarboxyl-terminal domain of the heavy chain (HC) of BoNT/C1 (strain 573), a fragment fromthe HC of BoNT/D (strain BVD/-3) or a fragment from the amino-terminal domain of theheavy chain (HN) of BoNT/C1 (strain 573) were expressed in Escherichia coli, andadministered as immunogens to mice. Monoclonal antibodies (mAbs) against therecombinant BoNT fragments were prepared by three fusions. MAbs recognising nativeBoNT/C1 and BoNT/D were detected by enzyme-linked immunosorbent assay (ELISA). Ninemonoclonal antibodies (mAbs) were produced, six of which recognised a BoNT fragmentthat is highly conserved across all serotype C and D producing strains. We conclude thatthese mAbs and this approach to mAb production may facilitate the development ofimmunological diagnostic techniques that are not constrained by the existence of mosaicisoforms for the detection and identification of serotypes C and D.

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