title: Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells--a potential mechanism for targeting liver anti-fibrotic therapeutics.
abstract: OBJECTIVE
  Hepatic stellate cells are pivotal to fibrogenesis in the liver and many potential anti-fibrotic therapeutics are required to act on targets within hepatic stellate cells. The aim of this study was to generate a human antibody fragment to hepatic stellate cells.
  
  
  METHODS
  Phage display was used to generate a human monoclonal antibody fragment to a peptide sequence present on an extracellular domain of synaptophysin, a protein expressed on the surface of hepatic stellate cells.
  
  
  RESULTS
  An antibody fragment was isolated (termed C1-3), expressed in bacteria and purified. Fluorescently-labelled C1-3 antibody associated with human hepatic stellate cells but not hepatocytes in culture. Binding of fluorescently labelled C1-3 to hepatic stellate cells was blocked by the extracellular synaptophysin peptide sequence and uptake of the antibody intracellularly was inhibited by monensin. The toxin tributyl tin-when conjugated to C1-3-retained the ability to kill hepatic stellate cells confirming that C1-3 is sequestered intracellularly.
  
  
  CONCLUSIONS
  This antibody fragment may be an effective means to target therapeutics to human hepatic stellate cells.
authors.affiliation: School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
authors.forename: Lucy J
authors.initials: LJ
authors.lastname: Elrick
authors.forename: Val
authors.initials: V
authors.lastname: Leel
authors.forename: Morgan G
authors.initials: MG
authors.lastname: Blaylock
authors.forename: Linda
authors.initials: L
authors.lastname: Duncan
authors.forename: Matthew R
authors.initials: MR
authors.lastname: Drever
authors.forename: Gillian
authors.initials: G
authors.lastname: Strachan
authors.forename: Keith A
authors.initials: KA
authors.lastname: Charlton
authors.forename: Matthew
authors.initials: M
authors.lastname: Koruth
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Porter
authors.forename: Matthew C
authors.initials: MC
authors.lastname: Wright
mesh_terms: D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000235:genetics*; D001483:Base Sequence; D001665:Binding Sites / Q000276:immunology; D022781:Hepatocytes / Q000276:immunology*; D006801:Humans; D007128:Immunoglobulin Fragments / Q000737:chemistry / Q000235:genetics*; D007167:Immunotherapy / Q000379:methods*; D066298:In Vitro Techniques; D008103:Liver Cirrhosis / Q000276:immunology* / Q000628:therapy; D008969:Molecular Sequence Data; D019151:Peptide Library; D017434:Protein Structure, Tertiary; D016708:Synaptophysin / Q000235:genetics / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D016708:Synaptophysin