Anti-Mouse CD279 (PD-1) [Clone RMP1-14] — Purified in vivo PLATINUM™ Functional Grade

Clone RMP1-14 is commonly used for in vivo blockade of PD-1 signaling in mice, particularly for research in cancer immunotherapy, immune regulation, and mechanistic studies of immune checkpoint pathways.

Key in vivo applications include:

• Cancer Immunotherapy Studies: RMP1-14 is extensively used in preclinical mouse models (such as syngeneic and genetically engineered models) to block PD-1/PD-L1 interactions, thereby enhancing anti-tumor immune responses, reducing tumor growth, and improving survival outcomes.
• Mechanistic Immunology: The antibody is used to investigate the role and regulation of the PD-1 pathway in T cell exhaustion, activation, and immune homeostasis.
• Combination Therapies and Tumor Model Benchmarking: RMP1-14 frequently serves as a benchmark or control antibody in studies comparing different immunotherapeutic strategies, including combinations with other immune checkpoint inhibitors or chemotherapies.
• Functional Immune Assays: Its application includes validating PD-1 blockade in vivo by monitoring T cell proliferation, cytokine production, and depletion studies, often via flow cytometry and immunohistochemistry.

Additional notes:

• Isotype and Mechanism: As a rat IgG2a monoclonal antibody, RMP1-14 can mediate antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), which may contribute to its efficacy in tumor models.
• Specificity: RMP1-14 is specific to mouse PD-1 and does not cross-react with human PD-1, making it unsuitable for translational studies in humanized mice where cross-reactivity is required.

In summary, RMP1-14 is a cornerstone tool for in vivo PD-1 blockade in murine studies, with principal utility in preclinical cancer immunotherapy and immune regulation research.

RMP1-14 is a commonly used monoclonal antibody against mouse PD-1, often utilized in immunotherapy research, particularly in in vivo models. Here are some other antibodies or proteins frequently used alongside RMP1-14:

1. 29F.1A12: This antibody is another anti-PD-1 monoclonal antibody often used for both in vivo and in vitro studies. It has a higher avidity than RMP1-14 and can effectively block PD-1/PD-L1 interactions.

2. J43: Similar to RMP1-14 and 29F.1A12, J43 is used for blocking PD-1 signaling. It is noted for its efficacy in tumor models and is used in both in vivo and in vitro research.

3. PD-L1 and PD-L2: These are the ligands for PD-1, and antibodies targeting them can also be used in studies involving PD-1. Commonly used antibodies include 10F.9G2 and MIH6 against PD-L1.

4. Other PD-1 Agonist Antibodies: While RMP1-14 exhibits both blocking and agonist activities, other antibodies like RMP1-30 are recognized for their agonist properties, binding to different regions of the PD-1 molecule.

Using these antibodies in combination with RMP1-14 can provide a comprehensive understanding of PD-1 mediated immune responses and their modulation in various experimental settings.

Clone RMP1-14 is a widely used anti-mouse PD-1 monoclonal antibody in immuno-oncology research, mainly known for its effectiveness in preclinical models to block the PD-1 pathway. Key findings from the scientific literature regarding RMP1-14 are as follows:

• Proven Efficacy in Tumor Models: RMP1-14 demonstrates significant anti-tumor activity in multiple mouse cancer models, including glioma, melanoma, breast cancer, and mesothelioma, especially when combined with other therapies. Tumor suppression rates range from 65% to over 80% in combination settings.

• Benchmark for PD-1 Blockade: RMP1-14 is considered a gold standard for in vivo PD-1 blockade studies due to its ability to enhance anti-tumor immune responses, reduce tumor growth, and improve survival outcomes in syngeneic and genetically engineered mouse models.

• High Specificity and Blocking Activity: It was developed for high affinity and specificity to mouse PD-1, selectively blocking the PD-1/PD-L1 and PD-1/PD-L2 interactions that suppress T cell function.

• Functional Role: RMP1-14 primarily acts as a blocking antibody in in vivo systems, efficiently blocking inhibitory signaling but exhibiting minimal agonistic (suppressive) activity, thus aligning with its therapeutic intent to reinvigorate T cell activity.

• Binding Affinity and Comparative Performance: Though effective, RMP1-14 has a lower binding avidity compared to some other anti-PD-1 clones such as 1A12; 1A12 outperforms RMP1-14 at lower concentrations due to higher avidity (IC~50~ for RMP1-14 is around 3–4 µg/mL, versus ~0.03 µg/mL for 1A12). However, RMP1-14 outperforms other clones like J43 in tumor growth inhibition and survival in certain models.

• Validated Protocols and Reproducibility: The extensive preclinical literature and well-established dosing protocols make RMP1-14 a reliable and reproducible tool for researchers.

• Antibody Characteristics: RMP1-14 is a rat IgG2a isotype, developed through hybridoma technology for functional PD-1 blockade in mice.

• Limitations and Considerations: Its lack of cross-reactivity with human PD-1 means it is not suitable for clinical or translational studies involving humanized models, where human-reactive antibodies like EH12.2H7 are preferred. Its lower avidity compared to some other clones can also affect performance in studies requiring minimal antibody concentrations.

• Mechanistic and Immunological Insights: Studies confirm that RMP1-14 increases cytokine production (e.g., IFN-γ) in T cells, restauring anti-tumor immunity by releasing PD-1-mediated inhibitory signals.

In summary, RMP1-14 is a validated and potent tool for in vivo mouse studies on PD-1 blockade, valued for its specificity, reproducibility, and well-documented anti-tumor effects, though its properties should be carefully matched to the experimental context.

Dosing regimens for the RMP1-14 clone, an anti-mouse PD-1 antibody, can vary across different mouse models primarily based on the specific experimental objectives and the characteristics of the models used. Here are some general guidelines and considerations:

Typical Dosing Range

• General Dosing Range: The typical dosing range for RMP1-14 is between 200-500 μg per mouse, administered via intraperitoneal (IP) injection every 3-4 days.

Factors Influencing Dosing Regimens

• Tumor Type and Model: The choice of tumor model, such as MC38 colon carcinoma or B16 melanoma, can influence the dosing regimen. Different tumor models may require adjustments in dosage or frequency to optimize the efficacy of PD-1 blockade.
• Immune System Competency: The competency of the mouse's immune system can affect how well the PD-1 blockade works. Models with intact immune systems may require specific dosing strategies to avoid adverse effects while maximizing anti-tumor response.
• Genetic Background: Variations in the genetic background of different mouse strains can influence their response to PD-1 blockade. Therefore, dosing may need to be adjusted based on the strain used in the study.

Troubleshooting and Adjustments

• Common Issues: Treatment responses can vary due to differences in tumor models or animal strains. Well-characterized models and consistent experimental conditions are crucial to mitigate variability.
• Adverse Effects: Monitoring for adverse effects like autoimmunity or toxicity is important. Strategies such as dose reduction or treatment discontinuation may be necessary.
• Anti-Drug Antibodies (ADAs): The development of ADAs against RMP1-14 can limit its efficacy over time. Intermittent dosing or immunomodulation may help reduce ADA formation.

Overall, while there is a standard dosing range for RMP1-14, the regimen can be tailored based on the specific needs and characteristics of individual mouse models.