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Related Experiment Video

Updated: Apr 1, 2026

Identifying PD-1/PD-L1 Inhibitors with Surface Plasmon Resonance Technology

Identifying PD-1/PD-L1 Inhibitors with Surface Plasmon Resonance Technology

Published on: May 2, 2025

A Bead-Based Screening Platform for Identifying Monoclonal Antibodies That Disrupt PD-1/PD-L1 Interactions.

Veridiane M Pscheidt¹, Rodrigo B Gassen¹, José E Sacconi Nunes¹

¹Immunotherapy Laboratory, Federal University of Health Sciences of Porto Alegre, Porto Alegre, Rio Grande do Sul, Brazil, ufcspa.edu.br.

Journal of Immunology Research

|March 31, 2026

Summary

A new bead-based flow cytometry assay quantifies programmed cell death protein 1 (PD-1)/PD-L1 blockade by antibodies. This reliable method supports immunotherapy development by screening PD-1/PD-L1 blocking antibodies effectively.

Keywords:

PD-L1 antibodies flow cytometry immune checkpoints inhibitors immunotherapy

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Author Spotlight: Magnetic Fluorescent Bead-Based Dual-Reporter Flow Analysis of PDL1-Vaxx Peptide Vaccine-Induced Antibody Blockade of the PD-1/PD-L1 Interaction

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Published on: July 7, 2023

Monitoring PD-1-Blocking Antibodies Bound to T Cells Derived from a Drop of Peripheral Blood

Monitoring PD-1-Blocking Antibodies Bound to T Cells Derived from a Drop of Peripheral Blood

Published on: February 5, 2020

Area of Science:

Immunology
Biotechnology
Cancer Research

Background:

Monoclonal antibodies (mAbs) targeting immune checkpoints like PD-1/PD-L1 are crucial in immunotherapy.
Scalable methods for assessing the functional blockade of PD-1/PD-L1 interactions are limited.

Purpose of the Study:

To develop and validate a bead-based flow cytometry assay for quantifying the inhibition of PD-1/PD-L1 interaction by antibodies.
To establish a reliable tool for screening PD-1/PD-L1 blocking antibodies.

Main Methods:

Conjugating recombinant human PD-1 to beads and measuring its interaction with fluorochrome-labeled recombinant human PD-L1.
Quantifying the inhibitory activity of anti-PD-L1 mAbs by their ability to disrupt the PD-1/PD-L1 interaction.
Validating the assay's precision, reproducibility, and functional correlation with T cell activation markers (CD69).

Main Results:

The assay demonstrated dose-dependent inhibition of PD-1/PD-L1 interaction by an anti-PD-L1 mAb, with a mean IC50 of 3.122 µg/mL.
The method showed high reproducibility (RSD < 20%) and successfully distinguished antibodies with different affinities.
The antibody concentration near the IC50 significantly restored T cell CD69 expression in a coculture assay.

Conclusions:

The developed bead-based flow cytometry assay is a reproducible and reliable tool for quantifying PD-1/PD-L1 antibody blocking activity.
This high-throughput compatible platform supports the discovery and development of novel immunotherapy agents targeting the PD-1/PD-L1 pathway.