Removal of N-Linked Glycosylation Enhances PD-L1 Detection and Predicts Anti-PD-1/PD-L1 Therapeutic Efficacy

Cancer Cell. 2019 Aug 12;36(2):168-178.e4. doi: 10.1016/j.ccell.2019.06.008. Epub 2019 Jul 18.

Abstract

Reactivation of T cell immunity by PD-1/PD-L1 immune checkpoint blockade has been shown to be a promising cancer therapeutic strategy. However, PD-L1 immunohistochemical readout is inconsistent with patient response, which presents a clinical challenge to stratify patients. Because PD-L1 is heavily glycosylated, we developed a method to resolve this by removing the glycan moieties from cell surface antigens via enzymatic digestion, a process termed sample deglycosylation. Notably, deglycosylation significantly improves anti-PD-L1 antibody binding affinity and signal intensity, resulting in more accurate PD-L1 quantification and prediction of clinical outcome. This proposed method of PD-L1 antigen retrieval may provide a practical and timely approach to reduce false-negative patient stratification for guiding anti-PD-1/PD-L1 therapy.

Keywords: PD-1; PD-L1; antibody-based detection; biomarker; glycosylation/deglycosylation; heterogeneity/homogeneity; immune checkpoint; immunohistochemistry; immunotherapy.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • A549 Cells
  • Antibodies / immunology*
  • Antibody Specificity
  • Antineoplastic Agents, Immunological / therapeutic use
  • B7-H1 Antigen / antagonists & inhibitors
  • B7-H1 Antigen / immunology
  • B7-H1 Antigen / metabolism*
  • Clinical Decision-Making
  • False Negative Reactions
  • Glycosylation
  • Humans
  • Immunohistochemistry*
  • Jurkat Cells
  • MCF-7 Cells
  • Neoplasms / drug therapy
  • Neoplasms / immunology
  • Neoplasms / metabolism*
  • Patient Selection
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / metabolism*
  • Predictive Value of Tests
  • Protein Processing, Post-Translational*
  • Reproducibility of Results
  • Specimen Handling / methods*
  • THP-1 Cells

Substances

  • Antibodies
  • Antineoplastic Agents, Immunological
  • B7-H1 Antigen
  • CD274 protein, human
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase