Detection and typing of herpes simplex viruses by using recombinant immunoglobulin fragments produced in bacteria

Paola Cattani, Rosaria Santangelo, Giovanni Fadda, Gian Maria Rossolini, Stefania Cresti, Dennis R. Burton, R. Anthony Williamson, Pietro P. Sanna

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11 Citazioni (Scopus)

Abstract

Thirty-seven bacterial clones producing human recombinant monoclonal antibody Fab fragments (rFabs) reactive to herpes simplex virus (HSV) antigens were selected from a human combinatorial antibody library constructed in a phage-display vector by a panning procedure against an HSV lysate. Thirty-four of the HSV-specific rFabs were able to specifically recognize HSV-infected cells in indirect immunofluorescence (IF) assays; of these, 25 recognized cells infected by either HSV type 1 (HSV-1) or HSV-2, while 9 recognized only HSV-1-infected cells. One HSV type-common rFab (rFab H37) and one HSV-1-specific rFab (rFab H85) were further evaluated as reagents for viral detection and typing by IF staining in 134 HSV-positive (72 HSV-1 and 62 HSV-2) viral cultures from clinical specimens. The results obtained with these two rFabs were fully consistent with those obtained with a commercial preparation of fluorescein-labeled anti-HSV type-specific murine monoclonal antibodies. The detection sensitivity with the type-common rFab in indirect IF assays was higher overall than that provided by the type-specific murine monoclonal antibodies. Preparations of rFabs suitable for IF staining can be easily and inexpensively obtained in a clinical microbiology laboratory from Escherichia coli cultures. Similar HSV-specific rFabs, therefore, could be advantageous for in vitro diagnostic purposes.
Lingua originaleEnglish
pagine (da-a)1504-1509
Numero di pagine6
RivistaJournal of Clinical Microbiology
Volume35
Stato di pubblicazionePubblicato - 1997

Keywords

  • Animals
  • Antibodies, Monoclonal
  • Antibodies, Viral
  • Antigens, Viral
  • Base Sequence
  • Cercopithecus aethiops
  • Escherichia coli
  • Fluorescent Antibody Technique, Indirect
  • Genetic Vectors
  • Herpesvirus 1, Human
  • Herpesvirus 2, Human
  • Humans
  • Immunoglobulin Fab Fragments
  • Molecular Sequence Data
  • Peptide Library
  • Recombinant Fusion Proteins
  • Sequence Analysis, DNA
  • Simplexvirus
  • Vero Cells

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