Catalog #CP085
RecombiMAb anti-mouse CD25
Clone
PC-61-CP085
Isotype
Mouse IgG2a, λ
(switched from Rat IgG1, λ)
(switched from Rat IgG1, λ)
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Product Description
PC-61-CP085 is a recombinant, Fc-engineered chimeric version of the original PC-61 (PC-61.5.3) antibody. The variable domain sequences are identical but the constant region sequences have been switched from rat IgG1, λ to mouse IgG2a, λ for optimal use in murine models. This Fc-engineered antibody is reported to deplete Treg cells within the blood, lymph nodes and tumor microenvironment, whereas the original hybrioma-derived rat antibody failed to show reduction of intratumoral Treg cells. The enhanced Treg depletion within the tumor microenvironment by the Fc-engineered antibody is attributed to increased engagement with activating murine Fc gamma receptors, overcoming resistance caused by intratumoral upregulation of the inhibitory receptor FcγRIIb. Additionally, species-matched chimeric antibodies exhibit regulated effector functions—including Fc receptor binding and complement activation—and result in less immunogenicity and formation of anti-drug antibodies (ADAs) than xenogenic antibodies in animal models. The highly controlled sequence and lack of genetic drift in recombinant antibodies provide more reliable and reproducible results over hybridoma derived antibodies.
The PC-61.5.3-CP085 monoclonal antibody reacts with mouse IL-2Rα also known as CD25, Ly-43, p55, or Tac. IL-2Rα is the 55 kDa ligand-binding subunit of the interleukin 2 receptor alpha chain. IL-2Rα expression is largely restricted to tumor-infiltrating Treg cells in mice. Alone, the IL-2Rα binds IL-2 with relatively low affinity however, when IL-2Rα associates with IL-2Rβ (CD122) and the common gamma chain (CD132) the complex binds IL-2 with high affinity. The PC-61.5.3 antibody has been shown to inhibit the binding of IL-2 to both the low and high affinity IL-2 receptor forms and block CD25‐dependent IL‐2 signaling.
References:
Arce Vargas, Frederick et al. “Fc-Optimized Anti-CD25 Depletes Tumor-Infiltrating Regulatory T Cells and Synergizes with PD-1 Blockade to Eradicate Established Tumors.” Immunity vol. 46,4 (2017): 577-586. doi:10.1016/j.immuni.2017.03.013
Huss, David J et al. “Anti-CD25 monoclonal antibody Fc variants differentially impact regulatory T cells and immune homeostasis.” Immunology vol. 148,3 (2016): 276-86. doi:10.1111/imm.12609
Solomon, Isabelle et al. “CD25-Treg-depleting antibodies preserving IL-2 signaling on effector T cells enhance effector activation and antitumor immunity.” Nature cancer vol. 1,12 (2020): 1153-1166. doi:10.1038/s43018-020-00133-0
The PC-61.5.3-CP085 monoclonal antibody reacts with mouse IL-2Rα also known as CD25, Ly-43, p55, or Tac. IL-2Rα is the 55 kDa ligand-binding subunit of the interleukin 2 receptor alpha chain. IL-2Rα expression is largely restricted to tumor-infiltrating Treg cells in mice. Alone, the IL-2Rα binds IL-2 with relatively low affinity however, when IL-2Rα associates with IL-2Rβ (CD122) and the common gamma chain (CD132) the complex binds IL-2 with high affinity. The PC-61.5.3 antibody has been shown to inhibit the binding of IL-2 to both the low and high affinity IL-2 receptor forms and block CD25‐dependent IL‐2 signaling.
References:
Arce Vargas, Frederick et al. “Fc-Optimized Anti-CD25 Depletes Tumor-Infiltrating Regulatory T Cells and Synergizes with PD-1 Blockade to Eradicate Established Tumors.” Immunity vol. 46,4 (2017): 577-586. doi:10.1016/j.immuni.2017.03.013
Huss, David J et al. “Anti-CD25 monoclonal antibody Fc variants differentially impact regulatory T cells and immune homeostasis.” Immunology vol. 148,3 (2016): 276-86. doi:10.1111/imm.12609
Solomon, Isabelle et al. “CD25-Treg-depleting antibodies preserving IL-2 signaling on effector T cells enhance effector activation and antitumor immunity.” Nature cancer vol. 1,12 (2020): 1153-1166. doi:10.1038/s43018-020-00133-0
Specifications
| Isotype | Mouse IgG2a |
|---|---|
| Recommended Isotype Control(s) | RecombiMAb mouse IgG2a isotype control, anti-hen egg lysozyme |
| Recommended Dilution Buffer | InVivoPure pH 7.0 Dilution Buffer |
| Immunogen | IL-2-dependent cytolytic mouse T cell clone B6.1 |
| Reported Applications |
Western Blot* Flow Cytometry* CD25‐dependent IL‐2 signaling blockade* In-vivo regulatory T cell depletion* Intratumoral Treg depletion* *Reported for the original rat IgG1, λ antibody |
| Formulation |
PBS, pH 7.0 Contains no stabilizers or preservatives |
| Endotoxin |
≤0.5EU/mg (≤0.0005EU/μg) Determined by LAL assay |
| Purity |
≥95% Determined by SDS-PAGE |
| Sterility | 0.2 µm filtration |
| Production | Purified from mammalian cell supernatant in an animal-free facility |
| Purification | Protein G |
| Molecular Weight | 150 kDa |
| Murine Pathogen Tests |
Ectromelia/Mousepox Virus: Negative Hantavirus: Negative K Virus: Negative Lactate Dehydrogenase-Elevating Virus: Negative Lymphocytic Choriomeningitis virus: Negative Mouse Adenovirus: Negative Mouse Cytomegalovirus: Negative Mouse Hepatitis Virus: Negative Mouse Minute Virus: Negative Mouse Norovirus: Negative Mouse Parvovirus: Negative Mouse Rotavirus: Negative Mycoplasma Pulmonis: Negative Pneumonia Virus of Mice: Negative Polyoma Virus: Negative Reovirus Screen: Negative Sendai Virus: Negative Theiler’s Murine Encephalomyelitis: Negative |
| Storage | The antibody solution should be stored at the stock concentration at 4°C. Do not freeze. |
| Need a Custom Formulation? | See All Antibody Customization Options |