Anti-Mouse CD309 (VEGFR2) [Clone DC101] — Purified in vivo GOLD™ Functional Grade

DC101 is a rat monoclonal antibody clone that targets vascular endothelial growth factor receptor 2 (VEGFR-2) and is extensively used in mouse studies for cancer research and immunotherapy applications.

Direct Antibody Treatment

DC101 is administered directly to tumor-bearing mice as an antiangiogenic therapy. The antibody works by binding to VEGFR-2 on endothelial cells, directly inhibiting angiogenesis and reducing tumor growth. In experimental settings, DC101 treatment results in decreased endothelial cells and microvessel density in tumors, effectively starving tumors of their blood supply.

In immune-competent FVB mice, DC101 treatment alone leads to tumor regression through a T cell-dependent mechanism. The antibody induces significant expansion of tumor-specific CD8+ T cells and can result in durable tumor regression. Notably, tumors continue to regress both during and after DC101 treatment, indicating sustained therapeutic effects.

CAR-T Cell Therapy Applications

DC101 is also used to engineer chimeric antigen receptor (CAR) T cells that target VEGFR-2-expressing cells. In these studies, T cells are genetically modified to express DC101-based CARs, creating DC101-CAR T cells. These engineered T cells can specifically recognize and respond to VEGFR-2-expressing tumor cells and endothelial cells.

The DC101-CAR T cells demonstrate antigen-specific responses, including proliferation and IFN-? secretion when exposed to VEGFR-2-expressing targets. These modified T cells have shown effectiveness against multiple types of established syngeneic tumors in mouse models.

Immune Response Modulation

The effectiveness of DC101 varies significantly depending on the immune status of the mouse model. In FVB mice (which lack immune tolerance to tumor antigens), DC101 induces robust T cell-dependent tumor regression with enhanced tumor-specific immune responses. However, in neu-N mice (which have preexisting immune tolerance), DC101 only slows tumor growth without inducing frank regression.

To overcome immune tolerance in tolerant mouse models, DC101 can be combined with other treatments such as regulatory T cell depletion using cyclophosphamide, which enhances vaccine-induced tumor regression responses. This approach demonstrates how DC101 can be integrated into combination immunotherapy strategies.

DC101 treatment enhances the presentation of heterogeneous tumor epitopes, potentially expanding the repertoire of immune recognition beyond single immunodominant epitopes. This broad immune activation contributes to its therapeutic efficacy in immunocompetent mouse models.

The correct storage temperature for sterile packaged clone DC101 (anti-mouse VEGFR-2 monoclonal antibody) is -20°C to -70°C, using a manual defrost freezer and avoiding repeated freeze-thaw cycles.

This storage condition maintains antibody stability for up to 12 months from the date of receipt. For shorter-term storage (up to 1 month), 2°C to 8°C may be acceptable, but for long-term preservation, freezing at -20°C to -70°C is specifically recommended. Always follow these precautions:

• Use a manual defrost freezer
• Avoid repeated freeze-thaw cycles, as this can degrade antibody quality

There is no indication in authoritative sources that clone DC101 should be stored at standard refrigerator (2–8°C) or room temperature for long-term use.

The most commonly used antibodies or proteins combined with DC101 (an anti-VEGFR2 antibody) in the literature include:

• Anti-VEGF-A antibody (e.g., 2G11-2A05): Frequently co-administered with DC101 to simultaneously block VEGF-A ligand and its receptor, enhancing anti-angiogenic effects in tumor models.
• Cetuximab (anti-EGFR antibody): Used in combination with DC101 to concurrently inhibit EGFR and VEGFR2 signaling, particularly in models of squamous cell carcinoma and other epithelial tumors.
• Chemotherapeutic agents (e.g., gemcitabine): DC101 is often paired with cytotoxic drugs to assess synergistic effects on inhibiting tumor growth, as noted in pancreatic cancer research.
• Other VEGFR pathway-related antibodies (e.g., anti-FLK2 (2A13), anti-FLK2 (23H7)): Used for mechanistic studies or as controls in experiments examining VEGF pathway inhibition.
• Secondary detection antibodies: Such as anti-rat IgG-biotin or rabbit polyclonal antibodies used in immunoprecipitation, immunoblotting, or detection assays alongside DC101.

Context and common combinations:

• The dual blockade of VEGF ligand (e.g., VEGF-A) and its receptor (VEGFR2) is a prominent strategy to intensify inhibition of angiogenesis in oncology research.
• EGFR inhibitors (like cetuximab) are often combined with DC101 to target both angiogenic and proliferative pathways, particularly in epithelial cancers.
• Chemotherapy plus anti-VEGFR2 therapy: Combined regimens evaluate whether anti-angiogenic therapies sensitize tumors to standard cytotoxic agents or reduce resistance.

These patterns reflect a strategy aimed at disrupting multiple complementary pathways to enhance anti-tumor efficacy. DC101 is often used not in isolation, but as part of a combinatorial approach with other targeted or conventional therapies in preclinical models.

Clone DC101, a monoclonal antibody targeting VEGFR-2 (also known as Flk-1 or KDR), is widely cited in scientific literature for its role in inhibiting angiogenesis and tumor growth. Key findings from DC101-related studies include:

• Inhibition of Tumor Angiogenesis and Growth: DC101 significantly reduces tumor vascularization and growth across diverse preclinical cancer models, including pulmonary metastases, mammary carcinoma (4T1), and melanoma (B16). Effects include substantial decreases in microvessel density (up to 90% reduction after 2 weeks), suppression of tumor cell proliferation, induction of tumor cell and endothelial cell apoptosis, and increased tumor necrosis.

• Survival Benefits: In murine models, DC101 treatment not only diminishes tumor growth, but extends survival—treated mice outlive control mice with markedly reduced tumor burdens, and some demonstrate complete absence of macroscopic metastases.

• Synergistic Effects in Combination Therapy: DC101 synergizes with other therapies, such as cetuximab (an anti-EGFR antibody), to further suppress tumor growth and metastasis, as demonstrated in squamous cell carcinoma of the tongue models. Combination therapies can significantly reduce regional lymph node metastasis rates compared to monotherapies.

• Immunomodulatory Actions: Beyond antiangiogenic effects, DC101 has been shown to induce T cell–dependent antitumor immune responses. In immunocompetent mice bearing highly immunogenic tumors, DC101 leads to T cell–mediated tumor regression, requiring both CD4+ and CD8+ T cells. The antitumor effect is lost if T cells are depleted. Notably, DC101 can potentiate responses to tumor vaccines, with combined therapy yielding stronger immune activation than either agent alone.

• Vascular and Stromal Remodeling: DC101 prompts rapid regression of preexisting blood vessels in tumors and reduces stromal protease expression, mechanisms thought to contribute to its anti-tumor action and potentially affect tumor microenvironment reorganization.

• VEGF Pathway Specificity: DC101 specifically neutralizes mouse VEGFR-2 signaling, permitting distinction between paracrine and autocrine VEGF/VEGFR-2 activity in experimental models. This has enabled deeper investigation into VEGF-dependent tumor and endothelial biology.

In summary, scientific literature identifies DC101 as a critical research tool for dissecting VEGFR-2's role in tumor growth, angiogenesis, immune activation, and therapeutic combinations, substantiating its dual capacity for direct tumor suppression and indirect enhancement of antitumor immunity.