Antibody-Linked Fluorogen-Activating Proteins for Antigen Detection and Cell Ablation

Bioconjug Chem. 2019 Jan 16;30(1):63-69. doi: 10.1021/acs.bioconjchem.8b00720. Epub 2018 Dec 26.

Abstract

We demonstrate selective labeling of cell surface proteins using fluorogen-activating proteins (FAPs) conjugated to standard immunoglobulins (IgGs). Conjugation was achieved with a polypeptide reagent comprised of an N-terminal photoactivatable Fc-binding domain and a C-terminal FAP domain. The resulting FAP-antibody conjugates were effective agents for protein detection and cell ablation in cultured mammalian cells and for visualizing cell-cell contacts using a tethered fluorogen assay. Because our approach allows FAP-antibody conjugates to be generated for most currently available IgGs, it should have broad utility for experimental and therapeutic applications.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Cell Adhesion
  • Cell Line
  • Cetuximab / metabolism*
  • ErbB Receptors / metabolism
  • Fluorescent Dyes / chemistry*
  • Humans
  • Membrane Proteins / metabolism*
  • Photosensitizing Agents / chemistry
  • Receptor, ErbB-2 / metabolism
  • Recombinant Fusion Proteins / metabolism
  • Trastuzumab / metabolism*

Substances

  • Fluorescent Dyes
  • Membrane Proteins
  • Photosensitizing Agents
  • Recombinant Fusion Proteins
  • EGFR protein, human
  • ERBB2 protein, human
  • ErbB Receptors
  • Receptor, ErbB-2
  • Trastuzumab
  • Cetuximab