Anti-Human OX40L (Oxelumab) [Clone R4930]

Anti-Human OX40L (Oxelumab) [Clone R4930]

Product No.: LT1300

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Product No.LT1300
Clone
R4930
Target
TNFSF4
Product Type
Biosimilar Recombinant Human Monoclonal Antibody
Alternate Names
TNFSF4; GP34; CD252; TXGP1; CD134L; TNLG2B
Isotype
Human IgG1κ
Applications
B
,
FA
,
FC

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Antibody Details

Product Details

Reactive Species
Human
Host Species
Human
Expression Host
HEK-293 Cells
FC Effector Activity
Active
Immunogen
Original antibody raised against Human OX40L
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 Oxelumab 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.
Additional Applications Reported In Literature ?
FA
B
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 Oxelumab. Clone R4930 binds to Human OX40L (TNFSF4). This product is for research use only.
Background
Oxelumab is a human monoclonal antibody designed for the treatment of autoimmune diseases.3 Oxelumab recognizes human OX40L (TNFSF4). OX40L is a member of the tumor necrosis family and is the ligand for OX40 . The OX40/OX40L interaction generates an optimal T cell response and plays a significant role in determining the amount of memory T-cells remaining after the immune response.1 Therapeutic treatments with antibodies against TNFSF can sometimes result in serious side effects.2 More research is needed to understand the precise molecular mechanism of TNF inhibition. This cost-effective, research-grade Anti-Human OX40L (Oxelumab) utilizes the same variable regions from the therapeutic antibody Oxelumab making it ideal for research projects.
Antigen Distribution
OX40L is expressed in various cell types including antigen presenting cells, T-cells, vascular endothelial cells, mast cells, and natural killer cells.
PubMed
NCBI Gene Bank ID
Research Area
Biosimilars
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Costimulatory Molecules
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Immuno-Oncology
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Immunology

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.

In a pharmacokinetic (PK) bridging ELISA for measuring drug concentration in serum samples, research-grade Oxelumab biosimilars can be used as calibration standards or reference controls by employing a well-established bioanalytical strategy. Here's a step-by-step explanation:

Use of Biosimilars as Calibration Standards

  1. Preparation of Standards: Biosimilar standards are prepared in human serum to serve as calibration curves. These standards are typically prepared across a range of concentrations to ensure linearity and accuracy in the assay.

  2. Assay Calibration: The biosimilar is used as the analytical standard to calibrate the PK assay. This involves constructing a standard curve with the biosimilar at known concentrations, allowing for the quantification of both the biosimilar and reference products in test samples.

  3. Bioanalytical Equivalence: The biosimilar is selected as the analytical standard after demonstrating bioanalytical equivalence with the reference product. This involves rigorous validation to ensure that the method can accurately measure both products without significant variability.

Incorporation into a PK Bridging ELISA

  1. ELISA Setup: In a PK bridging ELISA, plates are coated with a specific antigen or protein to which the drug (e.g., Oxelumab) binds. The biosimilar standards are premixed with biotin-labeled antibodies and pipetted into the wells.

  2. Competitive Binding: Oxelumab in the serum samples competitively binds with the pre-coated protein in the wells. The amount of Oxelumab bound is inversely proportional to the color developed after adding the substrate solution, which is measured to determine the concentration.

  3. Validation and Precision: The assay's precision and accuracy are validated through intra- and inter-assay comparisons to ensure that the results are reliable and consistent across different experiments.

By using biosimilars as calibration standards, researchers can ensure that the PK bridging ELISA provides accurate and reliable data for pharmacokinetic studies, facilitating the comparison of biosimilar and reference products in clinical trials.

Regulatory Considerations

The bioanalytical strategy for using biosimilars as calibration standards must adhere to regulatory guidelines, such as those provided by the FDA, which emphasize the importance of method validation and bioanalytical equivalence in PK studies.

When studying tumor growth inhibition and characterizing tumor-infiltrating lymphocytes (TILs) using anti-TNFSF4 antibodies, researchers typically utilize both syngeneic and humanized models. However, there is no specific mention of anti-TNFSF4 antibodies in the provided search results. Generally, these models are essential for understanding the immune interactions and efficacy of various immunotherapies.

Syngeneic Models

Description: Syngeneic models involve transplanting tumor cells from the same genetic background into mice, allowing for a fully functional immune system. These models are extensively used for evaluating the efficacy of immunotherapies, including checkpoint inhibitors and costimulatory therapies like OX40 or 4-1BB.

Use in Research: Syngeneic models such as MC38, CT26, and RENCA are widely used for studying immune responses and tumor interactions. They offer a platform to assess how different immunotherapies interact with the immune system to target cancer cells, providing insights into potential clinical outcomes.

Humanized Models

Description: Humanized models involve mice that have been engrafted with human cells, tissues, or organs. They are used to study human-specific interactions, such as the effects of human monoclonal antibodies on human tumors in a mouse setting.

Use in Research: Humanized models are particularly useful for studying antibodies like anti-TNFRSF family members, as they can mimic the human immune environment more closely than syngeneic models. Bispecific antibodies, for example, have been tested in humanized mice models to inhibit tumor growth by modulating immune responses.

For anti-TNFSF4 antibodies specifically, humanized models would be more relevant for studying the effects on human-specific immune interactions, while syngeneic models can provide insights into broader immune modulation effects relevant to TNFSF4 signaling pathways. However, the specific use of anti-TNFSF4 antibodies in these models is not detailed in the provided search results.

Based on the available research, there appears to be some confusion in the query regarding oxelumab's role in immune-oncology. Oxelumab is actually designed to block the OX40-OX40L pathway rather than activate it, and it's primarily being developed for inflammatory conditions like asthma rather than cancer treatment.

Understanding Oxelumab's Mechanism

Oxelumab is a human monoclonal antibody that inhibits the interaction between OX40L (CD252) and its receptor OX40 (CD134) on T cells. This blocking action dampens immune responses by preventing T-cell activation and proliferation, which makes it suitable for treating autoimmune diseases and conditions with inappropriate immune activation. This is fundamentally different from the agonistic OX40 approaches used in cancer immunotherapy.

OX40 Agonists in Cancer Research

In contrast to oxelumab's blocking approach, cancer immunotherapy research focuses on OX40 agonists that activate this pathway to enhance anti-tumor immunity. For example, ivuxolimab (PF-04518600) is a fully human IgG2 monoclonal antibody OX40 agonist that has shown promise in clinical trials. In a phase I study, ivuxolimab demonstrated anti-tumor activity with partial responses observed in melanoma and hepatocellular carcinoma patients, including those who had previously received checkpoint inhibitor therapy.

Combination Strategies in Immune-Oncology

Current research on combining checkpoint inhibitors focuses on targeting multiple pathways simultaneously to overcome individual therapy limitations. The rationale behind these combinations lies in their different mechanisms of action:

Anti-CTLA-4 and Anti-PD-1/PD-L1 Combinations: These combinations work through complementary mechanisms, with anti-CTLA-4 primarily acting in lymph nodes to restore T-cell induction and proliferation, while anti-PD-1 acts at tumor sites to prevent cytotoxic T-cell neutralization. The CheckMate 067 trial in advanced melanoma showed that patients with PD-L1-negative tumors particularly benefited from ipilimumab plus nivolumab combination therapy.

Anti-PD-1 and Anti-LAG-3 Combinations: Research has revealed distinct mechanisms between different combination approaches. Anti-PD-1/LAG-3 combinations require CD4 T-cells for their anticancer effects and work by decreasing regulatory T-cell activity while increasing CD4 helper T-cell activity, leading to CD8 T-cell activation. This differs from anti-PD-1/CTLA-4 combinations, which result in direct accumulation and activation of cytotoxic CD8 T-cells without requiring CD4 T-cell presence.

Current Research Limitations

The available research does not describe specific studies using oxelumab biosimilars in combination with other checkpoint inhibitors for cancer treatment. This is likely because oxelumab's immune-suppressive mechanism would be counterproductive to the immune-activating goals of cancer immunotherapy. Instead, researchers are exploring OX40 agonists in combination with established checkpoint inhibitors to enhance anti-tumor immune responses.

The field continues to evolve toward understanding how different checkpoint pathways can be strategically combined to maximize therapeutic benefit while managing the increased toxicity risks associated with combination immunotherapy approaches.

In the context of immunogenicity testing, an Oxelumab biosimilar could theoretically be used in a bridging ADA ELISA as either a capture or detection reagent to monitor a patient's immune response against the therapeutic drug. However, specific details about Oxelumab's use in such a context are not provided in the available search results. Generally, here's how biosimilars like Oxelumab might be used in bridging ADA ELISAs:

Principle of Bridging ADA ELISA

The bridging ELISA is a sensitive method for detecting anti-drug antibodies (ADAs). It involves capturing ADAs on a plate using biotinylated drug molecules (e.g., Oxelumab biosimilar) and then detecting these bound ADAs using labeled versions of the same drug (e.g., HRP-labeled Oxelumab biosimilar).

Steps to Use Oxelumab Biosimilar in Bridging ELISA

  1. Capture Reagent:

    • Biotinylation: Oxelumab biosimilar is biotinylated to allow it to bind to streptavidin-coated plates.
    • Plate Preparation: The biotinylated Oxelumab biosimilar is added to the wells of a streptavidin-coated microtiter plate, which captures it effectively.
  2. Sample Addition: Serum samples from patients are added to the plate, allowing any ADAs present in the sample to bind to the captured Oxelumab biosimilar.

  3. Detection Reagent:

    • Labeling: An HRP-labeled version of the Oxelumab biosimilar is prepared.
    • Detection: The HRP-labeled Oxelumab biosimilar is added to the plate. This labeled drug binds to any ADAs already bound to the captured drug, forming a "bridge" between the captured and labeled drugs.
  4. Detection and Analysis:

    • Chromogenic Substrate Addition: A chromogenic substrate (e.g., TMB) is added to the wells. The enzyme (HRP) converts the substrate into a colored product, which is directly proportional to the amount of ADAs present in the sample.
    • Quantification: The color intensity is measured using a spectrophotometer, and the ADA levels are quantified based on a standard curve.

This approach allows for the sensitive detection of ADAs against therapeutic drugs like Oxelumab, helping in monitoring the patient's immune response. However, the specific use of Oxelumab or its biosimilar in this context would need to be tailored according to the drug's properties and the clinical requirements.

References & Citations

1. Mahmood,T. and Yang, P. (2012) N Am J Med Sci. 4(11): 533–536
2. Spicer, P. and Runkel, L. (2019) Expert Opin Investig Drugs. 28(2):99-106
3. Heo, YS. et al. (2014) Bio Design 2(2):55-61
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Flow Cytometry

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