Anti-Human CD20 [Clone 2H7] — Purified in vivo PLATINUM™ Functional Grade

The clone 2H7 is a monoclonal antibody that specifically targets the human CD20 antigen, which is expressed on the surface of B cells. In studies involving humanized mice, this antibody is commonly used for:

• Depleting or Tracking B Cells: By binding to CD20 on human B cells in humanized mouse models, the 2H7 antibody can be used to study B-cell behavior or to selectively deplete B cells in specific experimental conditions.

• Flow Cytometric Analysis and Immunohistochemistry: The 2H7 antibody is also applied in flow cytometry for analyzing CD20 expression on human peripheral blood lymphocytes and in immunohistochemical staining of frozen tissue sections to detect CD20-positive cells.

• Therapeutic Models: While not directly used for therapeutic purposes in humans, studies in mice can help understand how targeting CD20 might be effective in treating B-cell lymphomas or leukemias by demonstrating the potential of anti-CD20 therapies.

Commonly Used Antibodies and Proteins Alongside 2H7

The monoclonal antibody 2H7 is specified as an anti-human CD20 reagent and is used extensively to identify and study B cells in both research and therapeutic contexts. It is also employed as a positive control in antibody panels.

In Immunological Research

Rituximab is one of the most frequently cited antibodies used in conjunction with 2H7 in the literature, particularly because both target the CD20 antigen on B cells. They are often compared for their binding affinities and epitope recognition, with studies detailing the subtle structural differences in how each interacts with CD20.

Isotype control antibodies are commonly used alongside 2H7 as negative controls to ensure specificity in experimental assays. These controls help distinguish specific binding (due to 2H7) from nonspecific interactions.

F8 is another antibody mentioned in contexts involving 2H7, such as the construction of chelating recombinant antibodies (CRAbs) for targeted therapy applications. However, F8 targets a different protein (fibronectin extra domain A, not CD20), so its co-usage with 2H7 is a technical (e.g., bioconjugation, targeting, or imaging) rather than a functional or analytical overlap.

In Microbiology and Infectious Disease

Interestingly, 2H7 is also the name of a monoclonal antibody targeting Staphylococcus aureus surface antigen A (SasA) in some microbiology literature. In these studies, isotype control antibodies are again used as negative controls to validate the specificity of 2H7 binding and its functional effects (e.g., opsonophagocytic killing).

Summary Table

Key Points

• Rituximab is used for comparative and analytical studies with 2H7, especially in B cell research and CD20 targeting therapies.
• Isotype control antibodies are standard negative controls in experiments using 2H7 to ensure specificity.
• F8 is co-used in specialized applications such as targeted therapy or imaging, but it targets a different antigen.
• In microbiology contexts, isotype control antibodies are again the primary companion to 2H7 for specificity controls.

There is no evidence in the provided literature that other B7 family proteins or unrelated antibodies/proteins are routinely used alongside 2H7 in mainstream immunological research. The most common companions are rituximab (for CD20 targeting), isotype controls (for specificity), and, in niche applications, F8 (for advanced targeting strategies).

Clone 2H7 is a widely cited mouse monoclonal antibody that specifically recognizes the human CD20 cell surface antigen, a 33–37 kDa non-glycosylated phosphoprotein expressed on most mature B cells and many B-cell malignancies. Key findings from scientific citations of clone 2H7 include:

• Epitope and Specificity:

  • Clone 2H7 recognizes an epitope mapped to the sequence YNCEPANPSEKNSPST, situated in the large extracellular loop of human CD20, and only binds CD20 in its native oligomeric form.
  • The specificity is restricted to human CD20 and closely related primates, and some reports mention weak expression on certain T cell subsets and follicular dendritic cells.

• Expression and Biological Function of CD20:

  • CD20 is expressed during pre-B cell development, is present on resting and activated B cells, and is lost prior to terminal differentiation into plasma cells.
  • CD20 appears to regulate B cell activation, calcium conductance, cell cycle progression, and B cell proliferation.
  • CD20 does not shed or become internalized after antibody binding, making it a stable therapeutic and diagnostic target.

• Technical Applications:

  • Clone 2H7 is extensively used for flow cytometry, immunoprecipitation, and immunohistochemistry. It produces strong surface staining and does not require cell permeabilization or fixation for standard flow cytometric analysis.
  • The antibody is validated for use in research on humans and some non-human primates.

• Therapeutic and Diagnostic Relevance:

  • CD20 serves as a major therapeutic target in B cell lymphoma and other B cell disorders. Damaged B cells targeted by anti-CD20 therapies (such as engineered chimeric or humanized forms of 2H7) can be replaced by progenitor cells that do not express CD20.
  • A chimeric 2H7 antibody (mouse variable domains + human constant regions) was found to mediate potent antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) with human effector cells, overcoming limitations of the original mouse 2H7.

• Research Insights:

  • Studies involving clone 2H7 contributed to understanding the molecular structure and signaling functions of CD20, its role in lipid raft association, and dynamic colocalization with the B cell antigen receptor during immune activation.
  • Clone 2H7, alongside other anti-CD20 antibodies such as rituximab, has been used to elucidate mechanisms of action, binding epitopes, and therapeutic potential.

In summary, citations of clone 2H7 in scientific literature consistently demonstrate its reliability as a tool for detecting and depleting human CD20+ cells, its foundational role in B cell research, and its relevance to therapeutic antibody design and cancer immunotherapy.

Overview

The dosing regimen of the anti-CD20 monoclonal antibody clone 2H7 (mouse IgG2b) in mouse models is not strongly standardized and can vary considerably depending on the specific mouse model, experimental objectives, and the balance between efficacy and toxicity.

General Dosing Guidance

• Typical Range: Dosing regimens for clone 2H7 typically span 100–250 µg per mouse, administered by intraperitoneal injection every 3–4 days.
• Rationale for Variation: The exact dose and schedule may be adjusted for different mouse models—such as humanized, immunodeficient, or disease-specific strains—to optimize the balance of efficacy and safety.
• Humanized Mice: While the search results did not specify unique dosing for 2H7 in humanized mice, humanized mouse models themselves vary widely in their construction and immune reconstitution (e.g., THX, Hu-NSG). The type and extent of human immune engraftment, as well as the presence of human target cells (e.g., CD20+ B cells), could influence both the required dose and the treatment outcome. However, no unique 2H7 dosing regimen for these models was detailed in the available literature.

Factors Influencing Dosing

• Mouse Strain: Immunodeficient mice, such as NOD-scid or NSG, may tolerate different dosing due to altered immune function. Their lower immune cell diversity might reduce the risk of immune-mediated toxicities but could also require higher doses for efficacy against xenografted human cells.
• Disease Model: In cancer or autoimmune models, disease progression and target cell load may necessitate dose adjustments.
• Objective: Prophylactic vs. therapeutic use, depletion kinetics, and desired degree of target cell clearance can all affect regimen selection.
• Safety: Overly aggressive depletion regimens may cause morbidity or mortality, especially in highly immunodeficient or humanized mice.

Considerations for Humanized Mouse Models

Humanized mice (e.g., THX or Hu-NSG) are increasingly used to model human immune responses, but the literature does not provide a consensus on 2H7 dosing for these specific strains. Researchers must empirically determine the optimal dose based on the degree of human immune cell engraftment, target antigen (human vs. mouse CD20), and the experimental endpoint.

Summary Table

Conclusion

Dosing of clone 2H7 in mouse models typically ranges from 100–250 µg per mouse every 3–4 days by intraperitoneal injection, but must be tailored to the specific mouse strain, disease model, and experimental objectives. For humanized or immunodeficient mice, regimens should be empirically optimized, as these models present unique challenges and opportunities for immunotherapy research. No standardized protocol for 2H7 in humanized mice was found in the current literature; thus, pilot studies are recommended to establish optimal dosing in these advanced models.