Anti-Human IL-2Rα (CD25) (Basiliximab) [Clone Hu107] — Dylight® 488

Anti-Human IL-2Rα (CD25) (Basiliximab) [Clone Hu107] — Dylight® 488

Product No.: LT311

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Product No.LT311
Clone
Hu107
Target
CD25
Product Type
Biosimilar Recombinant Human Monoclonal Antibody
Alternate Names
IL-2-RA; IL2-RA; TAC antigen; p55; CD25
Isotype
Human IgG1κ
Applications
FC
,
IF
,
IHC

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Select Product Size
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Antibody Details

Product Details

Reactive Species
Cynomolgus Monkey
Rhesus Monkey
Human
Host Species
Human
Expression Host
HEK-293 Cells
FC Effector Activity
Active
Immunogen
Human CD25
Product Concentration
0.2 mg/ml
Formulation
This DyLight® 488 conjugate is formulated in 0.01 M phosphate buffered saline (150 mM NaCl) PBS pH 7.4, 1% BSA and 0.09% sodium azide as a preservative.
Storage and Handling
This DyLight® 488 conjugate is stable when stored at 2-8°C. Do not freeze.
Regulatory Status
Research Use Only (RUO). Non-Therapeutic.
Country of Origin
USA
Shipping
Next Day 2-8°C
Excitation Laser
Blue Laser (493 nm)
Applications and Recommended Usage?
Quality Tested by Leinco
FC The suggested concentration for Basiliximab biosimilar antibody for staining cells in flow cytometry is ≤ 1.0 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 ?
IF
IHC
Additional Reported Applications For Relevant Conjugates ?
CyTOF®
WB
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 Basiliximab. Basiliximab recognizes human CD25. This product is for research use only.
Background
CD25, a 55 kD type I transmembrane glycoprotein, has been shown to play roles in lymphocyte differentiation, activation, and proliferation. Many resting memory T cells constitutively express IL2Rα. It functions as the receptor for HTLV-1, resulting in its expression on neoplastic cells in adult T cell lymphoma/leukemia. CD25 (sIL-2R) has been used to track disease progression. Some additional clinical applications include Chagas disease, a disease characterized by a decline of CD25 expression on immune cells, and Multiple sclerosis, in which treatments with mAbs target CD25. Anti-Human IL-2R alpha (CD25) (Basiliximab) utilizes the same variable regions from the therapeutic antibody Basiliximab making it ideal for research projects.
Antigen Distribution
IL-2Rα is expressed on activated mature T and B lymphocytes, during early stages of thymocytes development, pre-B cells, and in activated CD4+ memory T-lymphocytes.
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 Basiliximab biosimilars are used as analytical standards or controls in pharmacokinetic (PK) bridging ELISAs by serving as the reference material for constructing standard curves, calibrating assay responses, and enabling quantitative measurement of Basiliximab concentrations in serum samples.

Context and Supporting Details:

  • In PK bridging ELISA assays, a calibration standard is needed to accurately quantify drug concentration in samples.
  • Research-grade Basiliximab biosimilar is used to make a standard curve: the biosimilar is serially diluted to prepare standards at known concentrations, typically spanning the expected range of drug concentrations in serum. Biosimilar standards are often prepared in the same matrix (human serum) as study samples for matrix equivalency.
  • During assay development and validation, precision and accuracy of quantification using the biosimilar standard are established by measuring both the biosimilar and the originator reference product across the assay's working range. If the ELISA shows that the biosimilar and reference product yield analytically equivalent results, the biosimilar can be confidently used as the sole calibrator for ongoing measurements.
  • This single calibrator approach (using the biosimilar standard for both biosimilar and reference product quantification) is favored because it reduces inter-assay variation and eliminates the need for separate calibration curves for each product.
  • Research biosimilars, such as Basiliximab, are cost-effective and are validated for equivalent binding/activity relative to the originator drug, making them suitable for routine PK assay use in preclinical or clinical research.
  • Biosimilars can also be used as reference controls—for instance, as low, medium, and high QC samples—to monitor the performance of the ELISA through each run.

Key Steps for Use in PK Bridging ELISA:

  • The biosimilar is reconstituted (if lyophilized) and diluted to prepare a series of calibration standards.
  • Serum samples, standards, and controls are assayed in the same ELISA plate to interpolate sample concentrations from the standard curve.
  • Validation includes assessment of parallelism, accuracy, and reproducibility between the biosimilar and the originator reference product to justify exclusive use of one as the assay standard.

Summary Table: Basiliximab Biosimilar Use in PK ELISA

UseExplanation
Calibration StandardPrepares standard curves for quantitation; reconstituted and diluted to known concentrations in human serum.
Reference ControlUsed as QC samples at various concentrations to ensure reliability and consistency of assay performance.
Bridging ApproachAnalytical equivalence with the reference product is demonstrated; if met, the biosimilar standard is used exclusively for all quantifications.

In summary, research-grade Basiliximab biosimilars serve as accessible, validated calibration and reference controls in PK bridging ELISAs, ensuring consistent, reproducible, and regulatory-aligned quantification of Basiliximab drug levels in serum samples.

Standard flow cytometry protocols for validating CD25 (IL-2R alpha) expression or Basiliximab biosimilar binding capacity commonly use the conjugated antibody (e.g., PE or APC-labeled Basiliximab biosimilar) to stain cells expressing CD25, followed by quantitative analysis of cell surface expression levels.

Typical Protocol Steps:

  • Cell Preparation:

    • Harvest cells (e.g., PBMCs, transfected HEK293, C8166 cells) and wash twice with buffer such as PBS/1% BSA to minimize nonspecific binding.
  • Antibody Incubation:

    • Incubate cells (1 × 10^6 cells/mL is a common concentration) with the conjugated Basiliximab biosimilar (PE- or APC-labeled) for 30–60 minutes at 4°C.
    • Use isotype control antibodies labeled with the same fluorochrome for appropriate negative controls.
  • Washing:

    • Wash cells 2–3 times with buffer to remove unbound antibody.
  • Flow Cytometric Analysis:

    • Analyze stained cells on a flow cytometer, capturing fluorescence intensity associated with the conjugate, which directly corresponds to CD25 surface expression or antibody binding capacity.

Validation/Experimental Controls:

  • Positive Control: Use a cell line with known high CD25 expression (e.g., IL-2-activated PBMCs, transfected HEK293 cells).
  • Negative Control: Use unstained cells or cells stained with isotype control.
  • Blocking Studies: For binding capacity, pre-incubate with excess unconjugated Basiliximab or recombinant IL-2 to confirm specificity by competition/inhibition of fluorescent antibody binding.

Alternative Protocols for Non-Direct Conjugates:

  • If the Basiliximab biosimilar is unconjugated, stain first with the primary antibody, then use a fluorochrome-conjugated secondary antibody (e.g., APC-anti-human IgG).
  • For ligand (IL-2) binding assays, cells can be incubated with biotinylated IL-2, followed by competing Basiliximab biosimilar and then detected via APC-Streptavidin conjugate.

Sample Protocol for Directly Conjugated Basiliximab Biosimilar:

1. Resuspend 1 × 10^6 cells in 100 µL of PBS/1% BSA.2. Add 1–10 µg/mL PE-labeled Basiliximab biosimilar.3. Incubate for 30–60 minutes at 4°C in the dark.4. Wash cells twice with PBS/1% BSA.5. Resuspend cells and acquire data on flow cytometer (e.g., BD FACSCalibur).6. Analyze expression levels using FlowJo or similar software.

Key Parameters and Notes:

  • Antibody concentrations and incubation times should be titrated for optimal signal-to-noise ratio—manufacturer protocols usually offer starting recommendations, but optimization is recommended for each application.

  • Isotype controls are essential for background determination.

  • Quantitative analysis is based on mean fluorescence intensity (MFI) shifts between control and test samples.

  • Competition/Blocking assays can further characterize binding capacity and specificity.

References:

  • These protocols are adapted from published flow cytometry validation studies using Basiliximab and biosimilar antibodies targeting CD25, including direct staining and blocking experiments in both cell lines and primary human cells.

Analytical Assays for Biosimilarity

Biopharmaceutical companies typically use a comprehensive set of analytical assays to confirm the structural and functional similarity of a proposed biosimilar to the originator drug. These assays include:

  • Physicochemical Characterization: This involves assessing the primary, secondary, and higher-order structure of the biosimilar using techniques like mass spectrometry and chromatography. It also includes evaluating post-translational modifications.

  • Biological Assays: These are crucial for demonstrating the biological function of the biosimilar. They include binding assays, enzyme kinetics, cell-based assays, and potency assays.

  • Purity and Impurity Analysis: Rigorous comparison of purity and impurity profiles between the biosimilar and the reference product is essential. This includes analyzing aggregates and other product-related variants.

  • Orthogonal Methods: Multiple complementary techniques are used to better characterize the properties of each product, supporting the demonstration of high similarity or sensitively assessing potential differences.

Leinco Biosimilar in Studies

There is no specific information available about the "Leinco biosimilar" being used in these studies within the provided search results. However, the general principles of biosimilar development and characterization may still apply. Typically, biosimilars are compared to their reference products using the analytical methods mentioned above, regardless of the specific biosimilar manufacturer or product name.

If "Leinco" refers to a specific manufacturer or product line, it would follow these standard analytical approaches to establish biosimilarity with an originator drug. However, without specific details, it's difficult to provide further insight into how a "Leinco biosimilar" is used in these studies.

References & Citations

Flow Cytometry
IF
IHC

Certificate of Analysis

Formats Available

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