Anti-Human VEGF (Bevacizumab) – Fc Muted™

Anti-Human VEGF (Bevacizumab) – Fc Muted™

Product No.: LT405

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Product No.LT405
Clone
A4.6.1
Target
VEGF
Product Type
Biosimilar Recombinant Human Monoclonal Antibody
Alternate Names
Vascular Endothelial Growth Factor; VEGF-A; VEGFA; Vascular Permeability Factor; VPF
Isotype
Human IgG1κ
Applications
B
,
ELISA
,
FC
,
IP
,
N
,
WB

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Select Product Size
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Antibody Details

Product Details

Reactive Species
Human
Host Species
Human
Expression Host
HEK-293 Cells
FC Effector Activity
Muted
Immunogen
Recombinant human VEGF.
Product Concentration
≥ 5.0 mg/ml
Endotoxin Level
< 1.0 EU/mg as determined by the LAL method
Purity
≥95% by SDS Page
≥95% monomer by analytical SEC
Formulation
This biosimilar antibody is aseptically packaged and formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.2 - 7.4 with no carrier protein, potassium, calcium or preservatives added. Due to inherent biochemical properties of antibodies, certain products may be prone to precipitation over time. Precipitation may be removed by aseptic centrifugation and/or filtration.
Product Preparation
Recombinant biosimilar antibodies are manufactured in an animal free facility using only in vitro protein free cell culture techniques and are purified by a multi-step process including the use of protein A or G to assure extremely low levels of endotoxins, leachable protein A or aggregates.
Pathogen Testing
To protect mouse colonies from infection by pathogens and to assure that experimental preclinical data is not affected by such pathogens, all of Leinco’s recombinant biosimilar antibodies are tested and guaranteed to be negative for all pathogens in the IDEXX IMPACT I Mouse Profile.
Storage and Handling
Functional grade preclinical antibodies may be stored sterile as received at 2-8°C for up to one month. For longer term storage, aseptically aliquot in working volumes without diluting and store at ≤ -70°C. Avoid Repeated Freeze Thaw Cycles.
Regulatory Status
Research Use Only (RUO). Non-Therapeutic.
Country of Origin
USA
Shipping
2-8°C Wet Ice
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for Adalimumab biosimilar antibody for staining cells in flow cytometry is ≤ 0.25 μg per 106 cells in a volume of 100 μl. Titration of the reagent is recommended for optimal performance for each application.
WB
ELISA
Additional Applications Reported In Literature ?
B
N
IP
Each investigator should determine their own optimal working dilution for specific applications. See directions on lot specific datasheets, as information may periodically change.

Description

Description

Specificity
This non-therapeutic biosimilar antibody uses the same variable region sequence as the therapeutic antibody Bevacizumab. Bevacizumab recognizes both native and reduced human VEGF (isoform 165). This product is for research use only.
Background
Bevacizumab is a monoclonal antibody that specifically recognizes vascular endothelial growth factor (VEGF). VEGF is a growth factor that participates in angiogenesis, vasculogenesis, and endothelial cell growth. It facilitates endothelial cell proliferation, cell migration, and the permeabilization of blood vessels. In addition, VEGF inhibits apoptosis. Bevacizumab neutralizes the biological activity of VEGF by preventing the interaction of VEGF with its receptors on the surface of endothelial cells, resulting in the regression of tumor vascularization, normalization of remaining tumor vasculature, and inhibition of the formation of new tumor vasculature, thus inhibiting tumor growth.1 Anti-Human VEGF (Bevacizumab) utilizes the same variable regions from the therapeutic antibody Bevacizumab making it ideal for research projects.
Antigen Distribution
VEGF is widely expressed in the thyroid, prostate, and various other tissues.
PubMed
NCBI Gene Bank ID
Research Area
Biosimilars

Leinco Antibody Advisor

Powered by AI: AI is experimental and still learning how to provide the best assistance. It may occasionally generate incorrect or incomplete responses. Please do not rely solely on its recommendations when making purchasing decisions or designing experiments.

Research-grade Bevacizumab biosimilars are used as calibration standards or reference controls in pharmacokinetic (PK) bridging ELISA assays to quantitatively measure drug concentrations in serum samples. These biosimilars serve as standardized materials to establish calibration curves or to validate assay performance, ensuring the accurate measurement of Bevacizumab—or biosimilar—levels during PK bridging studies.

Key roles and procedures:

  • Calibration Standards:
    Research-grade biosimilar Bevacizumab is serially diluted to generate a standard curve in the ELISA, typically spanning a defined range of concentrations (e.g., 39.06 ng/mL to 2500 ng/mL). Known concentrations of the biosimilar are spiked into blank matrix (such as serum) and processed identically to test samples. The resulting signal (typically absorbance) is plotted against the concentration to create the calibration curve, which allows for the interpolation of unknown sample concentrations.

  • Reference Controls:
    Biosimilars also act as positive controls to assess assay specificity, sensitivity, and reproducibility, as their binding patterns and physicochemical properties have been extensively validated to match those of the originator product within defined tolerances. This ensures result comparability and reliability.

  • Bridging ELISA format:
    In the context of PK bridging studies, ELISA plates may be coated with capture reagents (e.g., recombinant VEGF or anti-idiotype antibody) that bind Bevacizumab from serum samples. Detection reagents, often distinct for capture of biosimilar or innovator forms, further ensure assay specificity. The assay must be validated to measure both innovator and biosimilar with comparable sensitivity to support PK comparability.

  • Regulatory compliance:
    The use of biosimilar standards is in line with regulatory guidance (e.g., FDA, EMA) for bioanalytical method validation and comparability exercises in biosimilar drug development.

  • Analytical comparability:
    The biosimilar’s critical quality attributes, like binding potency and charge heterogeneity (e.g., pI value), are confirmed to be highly comparable to those of the reference product, ensuring its appropriateness as a calibrator or control.

Summary of process in a typical PK ELISA:

  • Prepare serial dilutions of biosimilar standard in assay buffer.
  • Add standards and serum samples to ELISA plate coated with anti-Bevacizumab reagent.
  • Incubate and wash, then add detection antibody.
  • Measure optical density and construct standard curve from biosimilar standards.
  • Calculate unknown sample concentrations by referencing the standard curve.

Research-grade Bevacizumab biosimilars are thus essential to PK bridging ELISA workflows, functioning as calibration standards and reference controls to ensure accurate, reproducible, and regulatory-compliant quantification of drug concentrations in serum during biosimilar development and comparative PK studies.

The primary in vivo models for administering research-grade anti-VEGF antibodies to study tumor growth inhibition and characterize tumor-infiltrating lymphocytes (TILs) are syngeneic mouse models and, to a lesser extent, humanized mouse models.

Syngeneic Models:

  • Definition: Syngeneic models use tumor cells and hosts of the same mouse strain (typically immunocompetent), enabling high-fidelity study of both tumor growth and immune cell dynamics, including TILs.
  • Applications: Anti-VEGF antibodies are administered to test tumor inhibition and assess immune responses, since the model preserves normal host immunity, permitting the isolation and characterization of TIL populations.
  • Example Models:
    • C57BL/6 mice with murine ovarian carcinoma (ID8 cells), engineered to overexpress VEGF, have been specifically created to investigate VEGF's roles in angiogenesis, tumor immunity, and therapy responses.
    • Other commonly used syngeneic models include CT26 (colon), 4T1 (breast), B16 (melanoma), RENCA (renal), and LLC (lung).
  • Advantages: These models are optimal for studying immune–tumor interactions, mechanisms of immune evasion, and for characterizing TIL subsets and functions in the context of anti-VEGF therapy.

Humanized Models:

  • Definition: Humanized models are immunodeficient mice engrafted with human immune cells and tumors, allowing for the study of human TILs and responses to anti-VEGF therapies.
  • Applications: While less commonly used due to cost and complexity, they facilitate the evaluation of human-specific TIL populations and their modulation by anti-VEGF antibodies in vivo, especially when the research focus is on human immune mechanisms.
  • Challenges: The immune system reconstitution and tumor engraftment are more complex, and antibody cross-reactivity (human vs. murine VEGF) must be considered.

Xenograft Models:

  • While widely used for studying tumor growth inhibition by anti-VEGF antibodies, these models use immunodeficient mice and human tumor cells. Their limitations include the absence of a fully functional immune system, restricting detailed study of mouse TIL biology.

Summary Table: Model Comparison for Anti-VEGF/TIL Studies

Model TypeImmune SystemTumor SourceTIL CharacterizationAnti-VEGF Use
SyngeneicMouse (fully intact)Mouse cell linesMouse TILs (robust)Routine
HumanizedHuman (reconstituted)Human tumorsHuman TILs (limited)Specialized
XenograftImmunodeficientHuman cell linesPoor (mouse TILs lacking)Common for growth, poor for TILs

Conclusion: For immunological characterization of TILs in the context of anti-VEGF therapy, syngeneic mouse models remain the gold standard due to their robust immunocompetence and suitability for in vivo antibody administration. Humanized models are an alternative when human immune–tumor interactions are required, albeit with practical and technical limitations.

Researchers are exploring the use of bevacizumab, an anti-vascular endothelial growth factor (VEGF) monoclonal antibody, in combination with immune checkpoint inhibitors to enhance synergistic effects in treating cancers. While specific studies on bevacizumab biosimilars combined with checkpoint inhibitors like anti-CTLA-4 or anti-LAG-3 biosimilars are limited, the concept revolves around leveraging the immunomodulatory effects of VEGF pathway inhibition to complement the action of checkpoint inhibitors.

Combining Bevacizumab with Checkpoint Inhibitors

  1. Rationale: Bevacizumab inhibits angiogenesis by targeting the VEGF pathway, which can modulate the tumor microenvironment, potentially enhancing the efficacy of immune checkpoint inhibitors. This combination aims to create a more favorable environment for immune cells to target cancer cells.

  2. Bevacizumab with Nivolumab: Studies have shown that combining bevacizumab with the anti-PD-1 inhibitor nivolumab can be effective in certain cancers. For example, a phase 2 trial in relapsed ovarian cancer demonstrated activity, although the combination's efficacy varies depending on the cancer type and patient population.

  3. Potential Synergies: Theoretically, combining bevacizumab biosimilars with anti-CTLA-4 or anti-LAG-3 inhibitors could amplify immune responses by addressing different aspects of tumor-immune interactions. However, specific clinical trials focusing on these combinations are necessary to confirm efficacy and safety.

Challenges and Future Directions

  • Clinical Validation: The effectiveness of such combinations needs to be validated through rigorous clinical trials. Current studies often focus on specific cancer types or combinations, and expanding these to include biosimilars and other checkpoint inhibitors will be crucial.

  • Complexity of Immune-Onocology Models: The complexity of the tumor microenvironment and differing immune responses across patients require personalized approaches. This complexity demands advanced analytical tools and larger, more diverse patient populations in clinical trials.

  • Regulatory Considerations: For biosimilars, regulatory pathways allow for extrapolation of indications based on comprehensive analytical and limited clinical data. However, extensive clinical trials may still be necessary to prove synergistic effects with other agents.

Conclusion

While the specific use of bevacizumab biosimilars in combination with anti-CTLA-4 or anti-LAG-3 biosimilars is not yet well-documented, researchers are actively exploring combinations of anti-VEGF agents with checkpoint inhibitors to enhance cancer treatment outcomes. Future studies will need to focus on these specific combinations to fully understand their potential synergistic effects.

In the context of immunogenicity testing, a Bevacizumab biosimilar can be used as either a capture or detection reagent in a bridging ADA ELISA to monitor a patient's immune response against the therapeutic drug. Here's how it could be utilized:

Bridging ADA ELISA Protocol

The bridging ADA ELISA is a versatile assay format used to detect anti-drug antibodies (ADAs) against therapeutic drugs like Bevacizumab. This approach typically involves:

  1. Capture Reagent: Initially, a biotinylated form of the Bevacizumab biosimilar is immobilized on streptavidin-coated plates. This acts as the capture reagent, allowing the assay to specifically bind anti-drug antibodies in the patient's serum or plasma.

  2. Detection Reagent: For the detection step, a second, differently labeled Bevacizumab biosimilar (e.g., HRP-labeled) is used. This labeled reagent binds to the captured anti-drug antibodies, forming a bridging complex that allows for the detection of these antibodies.

  3. Detection Signal: The enzymatic activity of the labeled reagent is then detected using a substrate like tetramethylbenzidine (TMB), producing a colorimetric signal that is proportional to the concentration of anti-drug antibodies in the sample.

Use of Bevacizumab Biosimilar

Using a Bevacizumab biosimilar as a capture or detection reagent in a bridging ADA ELISA offers several advantages:

  • Specificity: The biosimilar ensures specificity to Bevacizumab, reducing non-specific binding and improving assay accuracy.
  • Sensitivity: The bridging format allows for high sensitivity in detecting both monovalent and bivalent anti-drug antibodies.
  • Drug Tolerance: Developing assays with high drug tolerance is crucial for Bevacizumab, as it is present in high concentrations in clinical samples, which can interfere with ADA detection.

However, challenges such as interference from endogenous VEGF and the need for innovative dissociation methods to remove tightly bound drug components must be addressed to optimize the assay's performance.

Customization and Optimization

Each laboratory should customize and implement the bridging ELISA protocol according to their specific requirements, including the selection of high-quality reagents and optimal assay conditions to ensure accurate and reliable results.

By using a Bevacizumab biosimilar in this context, researchers can effectively monitor a patient's immune response to Bevacizumab and assess the drug's immunogenicity, which is critical for evaluating treatment efficacy and safety.

References & Citations

1. Pazdur, R. et al. (2018) Clin Cancer Res. 24(18):4365-70.
B
Indirect Elisa Protocol
Flow Cytometry
Immunoprecipitation Protocol
N
General Western Blot Protocol

Certificate of Analysis

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Disclaimer AlertProducts are for research use only. Not for use in diagnostic or therapeutic procedures.