Reverse proteomic antibody screening identifies anti adhesive VHH targeting VLA-3

Mol Immunol. 2009 Jun;46(10):2022-8. doi: 10.1016/j.molimm.2009.03.002. Epub 2009 Apr 8.

Abstract

Therapeutic approaches aimed at targeting tumor surface markers using monoclonal antibodies provide a powerful strategy in cancer treatment. Here we report selection of single variable domains (VHH) of llama heavy chain antibodies, using a VHH-phage-display library. A reverse proteomic approach was used to identify the cognate proteins recognized by enriched VHH on HeLa cells. One of these VHH bound the integrin alpha 3 beta 1 (VLA-3) and was further characterized. Most interestingly, this VHH could inhibit VLA-3 mediated cell-matrix adhesion. Our approach provides a fast and efficient method to screen for novel cell surface markers on normal and tumor cells that may find diagnostic or therapeutic application in disease management or treatment.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adhesiveness
  • Amino Acid Sequence
  • Animals
  • Antibodies / analysis*
  • Camelids, New World
  • Cell Adhesion
  • Cell Membrane / immunology
  • Enzyme-Linked Immunosorbent Assay
  • Extracellular Matrix / immunology
  • Fluorescent Antibody Technique
  • HeLa Cells
  • Humans
  • Immunoglobulin Heavy Chains / chemistry
  • Immunoglobulin Heavy Chains / immunology*
  • Immunoglobulin Variable Region / chemistry
  • Immunoglobulin Variable Region / immunology*
  • Immunoprecipitation
  • Integrin alpha3beta1 / chemistry
  • Integrin alpha3beta1 / immunology*
  • Keratinocytes / cytology
  • Keratinocytes / immunology
  • Microscopy, Fluorescence
  • Molecular Sequence Data
  • Proteomics / methods*
  • Reproducibility of Results

Substances

  • Antibodies
  • Immunoglobulin Heavy Chains
  • Immunoglobulin Variable Region
  • Integrin alpha3beta1