Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells - a potential mechanism for targeting liver anti-fibrotic therapeutics

L. J. Elrick, Valerie Hilda Marie Duncan Leel, Morgan Graeme Blaylock, Linda Duncan, Matthew Ross Drever, G. Strachan, Keith Alan Charlton, M. Koruth, Andrew Justin Radcliffe Porter, Matthew Christopher Wright

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Background/Aims: 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. (C) 2005 European Association. for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

Original languageEnglish
Pages (from-to)888-896
Number of pages8
JournalJournal of Hepatology
Volume42
Issue number6
DOIs
Publication statusPublished - Jun 2005

Keywords

  • liver
  • fibrosis
  • phage display
  • stellate
  • synaptophysin
  • PHAGE DISPLAY
  • RAT-LIVER
  • MANNOSE 6-PHOSPHATE
  • ESCHERICHIA-COLI
  • IN-VITRO
  • FIBROSIS
  • SYNAPTOPHYSIN
  • EXPRESSION
  • METYRAPONE
  • RESOLUTION

Cite this

Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells - a potential mechanism for targeting liver anti-fibrotic therapeutics. / Elrick, L. J.; Leel, Valerie Hilda Marie Duncan; Blaylock, Morgan Graeme; Duncan, Linda; Drever, Matthew Ross; Strachan, G.; Charlton, Keith Alan; Koruth, M.; Porter, Andrew Justin Radcliffe; Wright, Matthew Christopher.

In: Journal of Hepatology, Vol. 42, No. 6, 06.2005, p. 888-896.

Research output: Contribution to journalArticle

Elrick, L. J. ; Leel, Valerie Hilda Marie Duncan ; Blaylock, Morgan Graeme ; Duncan, Linda ; Drever, Matthew Ross ; Strachan, G. ; Charlton, Keith Alan ; Koruth, M. ; Porter, Andrew Justin Radcliffe ; Wright, Matthew Christopher. / Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells - a potential mechanism for targeting liver anti-fibrotic therapeutics. In: Journal of Hepatology. 2005 ; Vol. 42, No. 6. pp. 888-896.
@article{cac034b897934d1481ae3ed63d879191,
title = "Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells - a potential mechanism for targeting liver anti-fibrotic therapeutics",
abstract = "Background/Aims: 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. (C) 2005 European Association. for the Study of the Liver. Published by Elsevier B.V. All rights reserved.",
keywords = "liver, fibrosis, phage display, stellate, synaptophysin, PHAGE DISPLAY, RAT-LIVER, MANNOSE 6-PHOSPHATE, ESCHERICHIA-COLI, IN-VITRO, FIBROSIS, SYNAPTOPHYSIN, EXPRESSION, METYRAPONE, RESOLUTION",
author = "Elrick, {L. J.} and Leel, {Valerie Hilda Marie Duncan} and Blaylock, {Morgan Graeme} and Linda Duncan and Drever, {Matthew Ross} and G. Strachan and Charlton, {Keith Alan} and M. Koruth and Porter, {Andrew Justin Radcliffe} and Wright, {Matthew Christopher}",
year = "2005",
month = "6",
doi = "10.1016/j.jhep.2005.01.028",
language = "English",
volume = "42",
pages = "888--896",
journal = "Journal of Hepatology",
issn = "0168-8278",
publisher = "Elsevier",
number = "6",

}

TY - JOUR

T1 - Generation of a monoclonal human single chain antibody fragment to hepatic stellate cells - a potential mechanism for targeting liver anti-fibrotic therapeutics

AU - Elrick, L. J.

AU - Leel, Valerie Hilda Marie Duncan

AU - Blaylock, Morgan Graeme

AU - Duncan, Linda

AU - Drever, Matthew Ross

AU - Strachan, G.

AU - Charlton, Keith Alan

AU - Koruth, M.

AU - Porter, Andrew Justin Radcliffe

AU - Wright, Matthew Christopher

PY - 2005/6

Y1 - 2005/6

N2 - Background/Aims: 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. (C) 2005 European Association. for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

AB - Background/Aims: 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. (C) 2005 European Association. for the Study of the Liver. Published by Elsevier B.V. All rights reserved.

KW - liver

KW - fibrosis

KW - phage display

KW - stellate

KW - synaptophysin

KW - PHAGE DISPLAY

KW - RAT-LIVER

KW - MANNOSE 6-PHOSPHATE

KW - ESCHERICHIA-COLI

KW - IN-VITRO

KW - FIBROSIS

KW - SYNAPTOPHYSIN

KW - EXPRESSION

KW - METYRAPONE

KW - RESOLUTION

U2 - 10.1016/j.jhep.2005.01.028

DO - 10.1016/j.jhep.2005.01.028

M3 - Article

VL - 42

SP - 888

EP - 896

JO - Journal of Hepatology

JF - Journal of Hepatology

SN - 0168-8278

IS - 6

ER -