Related Experiment Video
Updated: Jan 19, 2026
Identifying PD-1/PD-L1 Inhibitors with Surface Plasmon Resonance Technology
Published on: May 2, 2025
Blocking the PD-1-PD-L1 axis by a novel PD-1 specific nanobody expressed in yeast as a potential therapeutic for
Zongshu Xian1, Linlin Ma2, Min Zhu3
1School of Pharmacy, Fudan University, Shanghai, China.
Abstract:
PD-1/PD-L1 pathway blocking with antibodies offers a vital and efficient therapeutic strategy to restore T cell-associated antitumor immunity and treats a variety of cancers in clinic. Nanobodies (Nbs) give several advantages over conventional monoclonal antibodies such as size, solubility, stability and costs. Additionally, P. pastoris is a suitable host for Nb production. Herein, we aim to produce and evaluate anti-PD-1 Nb derived from the P. pastoris. Our findings indicated that we successfully established the Nbs phage-displayed library against PD-1 with qualified library capacity and insert ratio. Anti-PD-1 Nb Nb97 was screened through PE-ELISA and flow cytometry. To extend half-life of Nb97, we contracted pPICZɑA-Nb97-Nb97-HSA recombination vector, which was then transformed into the system of P. pastoris X-33. The yield of purified Nb97-Nb97-Human serum albumin (HSA) fused protein (MY2935) reached to 2.3 g/L after 147 h of fermentation. Meanwhile, the blocking effect of MY2935 is similar to that of MY2626 (humanized Nb97-Fc), and MY2935 showed better performance on stimulating the immune function through PD-1 reporter assay. Hence, P. pastoris X-33 expressing and secreting functional anti-PD-1 Nb-HSA fusion protein might be a system of high yield and low cost.
Insights
This study developed anti-PD-1 nanobodies (Nbs) using P. pastoris, a yeast host. The resulting fusion protein, MY2935, effectively blocks the PD-1 pathway and stimulates anti-tumor immunity.
Area of Science:
- Immunology
- Biotechnology
- Molecular Biology
Background:
- The PD-1/PD-L1 pathway is a crucial target for cancer immunotherapy.
- Nanobodies (Nbs) offer advantages over traditional monoclonal antibodies for therapeutic applications.
- Pichia pastoris is a well-established host for recombinant protein production.
Purpose of the Study:
- To produce and characterize anti-PD-1 nanobodies (Nbs) derived from P. pastoris.
- To develop a fusion protein with an extended half-life for enhanced therapeutic potential.
- To evaluate the efficacy of the anti-PD-1 Nb-HSA fusion protein in blocking PD-1 and stimulating anti-tumor immunity.
Main Methods:
- Construction and screening of a phage-displayed nanobody library against PD-1.
- Engineering and expression of an anti-PD-1 Nb-Human Serum Albumin (HSA) fusion protein (MY2935) in P. pastoris X-33.
- Assessing the binding affinity, blocking capability, and immune-stimulating function of MY2935.
Main Results:
- A high-quality phage-displayed nanobody library against PD-1 was successfully constructed.
- The anti-PD-1 Nb-HSA fusion protein MY2935 was produced with a yield of 2.3 g/L in P. pastoris.
- MY2935 demonstrated comparable PD-1 blocking activity to a humanized antibody and enhanced immune stimulation in reporter assays.
Conclusions:
- P. pastoris X-33 is a suitable and cost-effective host for high-yield production of functional anti-PD-1 Nb-HSA fusion proteins.
- The developed anti-PD-1 nanobody fusion protein shows promise as a therapeutic agent for cancer immunotherapy.
- This study highlights the potential of nanobodies and P. pastoris expression systems in advancing cancer treatment strategies.
More Related Videos
10:18Author Spotlight: Magnetic Fluorescent Bead-Based Dual-Reporter Flow Analysis of PDL1-Vaxx Peptide Vaccine-Induced Antibody Blockade of the PD-1/PD-L1 Interaction
Published on: July 7, 2023
06:07Monitoring PD-1-Blocking Antibodies Bound to T Cells Derived from a Drop of Peripheral Blood
Published on: February 5, 2020
Related Concept Videos
07:04Identifying PD-1/PD-L1 Inhibitors with Surface Plasmon Resonance Technology
10:18Magnetic Fluorescent Bead-Based Dual-Reporter Flow Analysis of PDL1-Vaxx Peptide Vaccine-Induced Antibody Blockade of the PD-1/PD-L1 Interaction
06:07Monitoring PD-1-Blocking Antibodies Bound to T Cells Derived from a Drop of Peripheral Blood
09:11Y-90 Radioembolization and PD-1 Inhibitor as Neoadjuvant Treatment in Hepatocellular Carcinoma
06:03Orthotopic Transplantation of Syngeneic Lung Adenocarcinoma Cells to Study PD-L1 Expression
07:43Using 22C3 Anti-PD-L1 Antibody Concentrate on Biopsy and Cytology Samples from Non-small Cell Lung Cancer Patients