Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells--a potential mechanism for targeting liver anti-fibrotic therapeutics.
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.
Elrick
Lucy J
LJ
School of Medical Sciences, Institute of Medical Sciences, University of Aberdeen, Foresterhill, Aberdeen AB25 2ZD, UK.
Leel
Val
V
Blaylock
Morgan G
MG
Duncan
Linda
L
Drever
Matthew R
MR
Strachan
Gillian
G
Charlton
Keith A
KA
Koruth
Matthew
M
Porter
Andrew J
AJ
Wright
Matthew C
MC
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
D000911:Antibodies, Monoclonal; D007128:Immunoglobulin Fragments; D019151:Peptide Library; D016708:Synaptophysin