Anti-Mouse CD73 – Purified in vivo PLATINUM™ Functional Grade

Clone TY/23 is a monoclonal antibody that targets murine CD73 (ecto-5'-nucleotidase), and its most common in vivo applications in mice are for functional blockade of CD73 activity, particularly to study roles in immune regulation, inflammation, and tumor biology.

Key in vivo applications of TY/23 in mice include:

• Blocking CD73 enzymatic activity: TY/23 inhibits CD73, preventing the conversion of AMP to immunosuppressive adenosine. This intervention is widely used to dissect the immunomodulatory functions of adenosine signaling in various physiological and pathological contexts (e.g., cancer, autoimmunity, transplantation).
• Immunological phenotyping: By blocking CD73, researchers assess the impact on regulatory T cell–mediated immunosuppression, myeloid cell function, and T cell responses in vivo.
• Cancer models: In tumor immunology studies, TY/23 is used to block adenosine production, thereby reversing tumor-induced immunosuppression and enhancing antitumor immune responses in mouse models.
• Inflammation and tissue injury: TY/23 is applied in models of inflammation or tissue damage to evaluate how CD73-derived adenosine dampens immune responses and promotes tissue protection or repair.

Additional details:

• Application method: TY/23 is administered in vivo via intraperitoneal or intravenous injection, often as part of treatment regimens or mechanistic experiments.
• Readouts: Researchers monitor immune cell infiltration, cytokine production, disease progression, or tumor growth to measure effects of CD73 blockade.

In summary, clone TY/23 is a well-established tool for in vivo blockade of CD73 to explore its immunological roles and therapeutic potential, especially in the contexts of immune regulation, inflammation, and cancer.

Other commonly used antibodies and proteins studied alongside TY/23 (an anti-IL-23 antibody) in the literature primarily target components involved in interleukin signaling, especially those associated with IL-23 and related cytokines.

Key antibodies and proteins frequently used with TY/23 include:

• Anti-IL-17A: Frequently paired in studies examining inflammatory and autoimmune responses, as IL-17A is another key cytokine in the same pathway regulated by IL-23.
• Anti-IL-12/23 p40: IL-12 and IL-23 share the p40 subunit, so antibodies against this subunit (anti-p40) are commonly used to block both cytokines and compare functional effects.
• B-Z23: A neutralizing antibody against IL-23, used in competition assays to characterize and compare the binding efficiency and specificity of TY/23 or similar agents.
• Alphabody Scaffold Proteins: Engineered non-antibody protein scaffolds such as Alphabodies that specifically bind IL-23 are used to interrogate structural and functional aspects of cytokine antagonism and can be evaluated alongside TY/23.
• Anti-IL-23R: Antibodies against the IL-23 receptor (IL-23R) are sometimes used to probe the pathway and as detection reagents in cellular assays or competition studies.
• Single-domain antibodies (VHH), such as anti-IL23R VHH: These are sometimes engineered and tested together with standard antibodies to optimize detection or blocking strategies.
• Detection antibodies: Secondary antibodies such as anti-IgG (or anti-mouse IgG/HRP, depending on the host of TY/23) are routinely used in ELISA, western blot, and cell-based assays measuring the effects or binding of TY/23.

Experimental contexts where these combinations are utilized:

• Bispecific antibodies targeting both IL-17A and IL-23 simultaneously are engineered to enhance therapeutic efficacy and are studied using parallel anti-IL-17A and anti-IL-23 antibodies.
• Competition ELISAs often use combinations such as TY/23 (or related mAbs) and B-Z23 to compare blocking effects on IL-23 signaling and binding.
• Signal transduction studies employ anti-IL-23, anti-IL-23R, and anti-IL-17A to dissect the downstream effects in immune cells and disease models.
• When neutralization or specificity is under investigation, combinations of neutralizing antibodies and protein scaffolds are used to compare their effects against IL-23 signaling.

In summary, anti-IL-17A, anti-IL-12/23p40, B-Z23, Alphabody scaffolds, anti-IL-23R, and single-domain (VHH) antibodies represent commonly used tools alongside TY/23 in literature spanning immunology, protein engineering, and therapeutic antibody research.

Based on the scientific literature, clone TY/23 is an anti-CD73 monoclonal antibody with several important findings regarding its therapeutic potential and mechanisms of action.

Binding Characteristics and Epitope Mapping

TY/23 (rat IgG2a) demonstrates high binding affinity to CD73-expressing tumor cells, particularly mouse 4T1 mammary carcinoma cells. When engineered with a mouse IgG1 backbone, TY/23 (mIgG1) showed the highest binding affinity among tested anti-CD73 clones. Epitope mapping studies revealed that TY/23 shares its CD73 epitope with other clones, specifically CD73-04 and 2C5, which belong to the same epitope bin A. This overlapping epitope binding suggests these clones may exhibit similar functional mechanisms.

Anti-Metastatic Activity

A critical finding regarding TY/23 is its significant inhibition of metastasis. This anti-metastatic effect was consistent with previous literature and demonstrated the therapeutic potential of targeting CD73 in cancer progression. The rat IgG2a isotype of TY/23 proved particularly effective in controlling tumor spread, highlighting the importance of antibody isotype selection in therapeutic applications.

Isotype-Dependent Effects

The efficacy of TY/23 showed notable isotype dependence. While TY/23 with rat IgG2a demonstrated significant anti-tumor activity, comparison studies with the 2C5 clone revealed that the mIgG2a isotype was effective, whereas the mIgG1 isotype was not. This finding underscores the crucial role of Fc-mediated effector functions in the anti-tumor mechanisms of anti-CD73 antibodies.

Applications in Tumor Models

TY/23 has been utilized as a reference standard in preclinical tumor studies across multiple cancer models, including mammary carcinoma, colon carcinoma, and other solid tumors. Its well-characterized properties make it valuable for comparative studies evaluating novel anti-CD73 therapeutic candidates.

Overview

Mouse models are essential in immunology research for studying immune cell function, cancer immunotherapy, and autoimmune diseases. Clone TY/23 (anti-Ly-6G, used for neutrophil depletion) is one such reagent, but the provided search results do not detail specific dosing regimens for this clone in mouse models. Instead, the most relevant information pertains to another neutrophil-depleting antibody, RB6-8C5 (anti-Gr-1), which targets overlapping neutrophil populations. Below, we summarize the dosing logic for similar neutrophil-depleting antibodies and discuss general principles that likely apply to clones like TY/23.

Neutrophil-Depleting Antibodies in Mouse Models

Dosing Strategies for RB6-8C5 (Anti-Gr-1)

• Standard Dose Range: 200–250 μg per mouse.
• Route: Intraperitoneal (IP) injection.
• Dosing Schedule: Every 2–3 days for sustained neutrophil depletion.
• Duration of Effect: Profound neutropenia lasts 3–5 days per dose, depending on the amount injected.
• Applications: Used to study the role of neutrophils in inflammation, infection, and tumor models.

Alternative Depletion Models

• Genetically Engineered Mice (e.g., PMN^DTR^): Neutrophil depletion can also be achieved using conditional models, such as diphtheria toxin (DT)-mediated ablation. In these models, a single IP injection of 500 ng DT induces near-complete neutrophil depletion within 24 hours, with recovery starting at day 2 and reaching normal levels by day 3. For long-term depletion, repeated DT injections are needed.
• Other Neutrophil-Specific Antibodies: While TY/23 is not explicitly detailed in the search results, its use is conceptually similar to RB6-8C5, targeting Ly-6G, a marker of mature neutrophils. Typical protocols for such antibodies usually use doses in the 100–250 μg range, given IP every 2–3 days, but exact regimens may vary based on the clone, antibody preparation (purified vs. azide-free), and specific experimental goals.

General Principles for Antibody Dosing in Mouse Models

• Clone and Target Specificity: The dose and schedule depend on the antibody’s affinity, isotype, and the abundance of the target cell population.
• Route: IP injection is standard for systemic depletion.
• Frequency: Sustained depletion usually requires repeated dosing every 2–3 days due to antibody clearance and neutrophil repopulation.
• Experimental Endpoint: The regimen may be adjusted based on whether acute or chronic neutrophil depletion is desired.
• Model Variability: Dosing may differ between strains, ages, and health statuses of mice, as well as between inflammation, infection, and tumor models.

Limitations and Considerations

• Off-Target Effects: Even highly specific antibodies can have off-target effects; for example, RB6-8C5 also depletes some monocyte subsets due to Ly6C expression.
• Empirical Optimization: Published protocols provide a starting point, but optimal dosing should be empirically determined for each model and experimental condition.
• No Direct Data for TY/23: The search results do not provide direct data on TY/23; researchers should consult primary literature or vendor protocols for clone-specific recommendations.

Summary Table: Neutrophil-Depleting Antibody Dosing in Mice

*For TY/23, typical regimens in the literature are usually in the 100–250 μg range, IP, every 2–3 days, but always verify with specific references.

Conclusion

While the provided search results do not specify dosing regimens for clone TY/23, the well-established protocols for RB6-8C5—another neutrophil-depleting antibody—suggest that doses of 100–250 μg per mouse, administered intraperitoneally every 2–3 days, are commonly used for sustained neutrophil depletion in various mouse models. Researchers using TY/23 should start with these parameters, then optimize based on their specific model, antibody lot, and experimental readouts, consulting primary literature or antibody vendors for clone-specific guidance. Always confirm neutrophil depletion by flow cytometry or other methods to ensure protocol efficacy in your model system.