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Updated: Feb 9, 2026

Genetic Encoding of a Non-Canonical Amino Acid for the Generation of Antibody-Drug Conjugates Through a Fast Bioorthogonal Reaction
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Genetic Encoding of a Non-Canonical Amino Acid for the Generation of Antibody-Drug Conjugates Through a Fast Bioorthogonal Reaction

Published on: September 14, 2018

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FRET Reagent Reveals the Intracellular Processing of Peptide-Linked Antibody-Drug Conjugates

Byoung-Chul Lee1, Cecile Chalouni2, Sophia Doll1,2

  • 1Department of Protein Chemistry , Genentech, Inc. , 1 DNA Way , South San Francisco , California 94080 , United States.

Bioconjugate Chemistry
|June 2, 2018
PubMed

Insights

This study reveals that cellular background impacts antibody-drug conjugate (ADC) processing and payload delivery. A novel FRET assay offers robust assessment of intracellular ADC activation for improved cancer therapy development.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Cancer Therapy

Background:

  • Antibody-drug conjugates (ADCs) are successful cancer therapeutics.
  • Intracellular trafficking and metabolism of ADCs remain incompletely understood.
  • Understanding ADC activation is crucial for optimizing drug development.

Purpose of the Study:

  • To investigate the intracellular processing and trafficking of ADCs.
  • To develop a method for assessing ADC activation within cells.
  • To explore the influence of cellular background on ADC mechanisms.

Main Methods:

  • Incorporation of fluorescence resonance energy transfer (FRET) reporter groups into ADC linkers.
  • Comparative analysis of antibody-FRET drug conjugate trafficking in diverse model cell lines.
  • Assessment of intracellular payload transport and linker cleavage.

Main Results:

  • Cellular background significantly influences antigen-mediated antibody processing.
  • Limited cytosolic transport of drug payload observed in some tumor cells, despite efficient linker cleavage.
  • The FRET assay demonstrated a facile and robust method for evaluating intracellular ADC activation.

Conclusions:

  • Cellular context is a critical factor in ADC efficacy.
  • The developed FRET assay is a valuable tool for studying ADC intracellular dynamics.
  • This research provides insights for the future design and development of novel ADCs for cancer treatment.

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