Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form
As of April 1st VIVO Scientific Profiles will no longer updated for faculty, and the link to VIVO will be removed from the library website. Faculty profile pages will continue to be updated via Interfolio. VIVO will continue being used behind the scenes to update graduate student profiles. Please contact helplib@scripps.edu if you have questions.
How to download citations from VIVO | Alternative profile options

Intradiabodies, bispecific, tetravalent antibodies for the simultaneous functional knockout of two cell surface receptors

Academic Article
uri icon
  • Overview
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Jendreyko, N.
  • Popkov, M.
  • Beerli, R. R.
  • Chung, J. H.
  • McGavern, Dorian
  • Rader, Christoph
  • Barbas III, Carlos

publication date

  • November 2003

journal

  • Journal of Biological Chemistry  Journal

abstract

  • The specific and high affinity binding properties of intracellular antibodies (intrabodies), combined with their ability to be stably expressed in defined organelles, provides powerful tools with a wide range of applications in the field of functional genomics and gene therapy. Intrabodies have been used to specifically target intracellular proteins, manipulate biological processes, and contribute to the understanding of their functions as well as for the generation of phenotypic knockouts in vivo by surface depletion of extracellular or transmembrane proteins. In order to study the biological consequences of knocking down two receptor-tyrosine kinases, we developed a novel intrabody-based strategy. Here we describe the design, engineering, and characterization of a bispecific, tetravalent endoplasmic reticulum (ER)-targeted intradiabody for simultaneous surface depletion of two endothelial transmembrane receptors, Tie-2 and vascular endothelial growth factor receptor 2 (VEGF-R2). Comparison of the ER-targeted intradiabody with the corresponding conventional ER-targeted single-chain antibody fragment (scFv) intrabodies demonstrated that the intradiabody is significantly more efficient with respect to efficiency and duration of surface depletion of Tie-2 and VEGF-R2. In vitro endothelial cell tube formation assays suggest that the bispecific intradiabody exhibits strong antiangiogenic activity, whereas the effect of the monospecific scFv intrabodies was weaker. These findings suggest that simultaneous interference with the VEGF and the Tie-2 receptor pathways results in at least additive antiangiogenic effects, which may have implications for future drug developments. In conclusion, we have identified a highly effective ER-targeted intrabody format for the simultaneous functional knockout of two cell surface receptors.

subject areas

  • Animals
  • Antibodies
  • Cell Division
  • Cell Line
  • Cell Membrane
  • Cell Survival
  • Cells, Cultured
  • Endoplasmic Reticulum
  • Endothelium, Vascular
  • Flow Cytometry
  • Gene Library
  • Humans
  • Immunohistochemistry
  • Kinetics
  • Microscopy, Fluorescence
  • Models, Molecular
  • Phenotype
  • Plasmids
  • Protein Binding
  • Rabbits
  • Receptor Protein-Tyrosine Kinases
  • Receptor, TIE-2
  • Recombination, Genetic
  • Surface Plasmon Resonance
  • Time Factors
  • Umbilical Veins
  • Vascular Endothelial Growth Factor Receptor-2
scroll to property group menus

Identity

International Standard Serial Number (ISSN)

  • 0021-9258

Digital Object Identifier (DOI)

  • 10.1074/jbc.M307002200

PubMed ID

  • 12947084
scroll to property group menus

Additional Document Info

start page

  • 47812

end page

  • 47819

volume

  • 278

issue

  • 48

©2022 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support