Anti-Human CD257 (BAFF) (Tabalumab) – Dylight® 488

Anti-Human CD257 (BAFF) (Tabalumab) – Dylight® 488

Product No.: LT1411

- -
- -
Product No.LT1411
Clone
Tabalumab
Target
BAFF
Product Type
Biosimilar Recombinant Human Monoclonal Antibody
Alternate Names
Tabalumab, CD257, BAFF, TNFSF13b, BLYS
Isotype
Human IgG1κ

- -
- -
Select Product Size
- -
- -

Antibody Details

Product Details

Reactive Species
Cynomolgus Monkey
Human
Rabbit
Host Species
Human
Expression Host
HEK-293 Cells
FC Effector Activity
Active
Immunogen
Original antibody was raised against soluble human BAFF.
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.
State of Matter
Liquid
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
2-8°C Wet Ice
Excitation Laser
Blue Laser (493 nm)
Additional Reported Applications For Relevant Conjugates ?
N
IP
WB
ELISA
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 Tabalumab. Tabalumab neutralizes soluble human, cynomolgus monkey, and rabbit BAFF. Additionally, Tabalumab neutralizes membrane-bound BAFF. This product is for research use only.
Background
Tabalumab is a human monoclonal anti-B-cell activating factor (BAFF) antibody intended for the treatment of autoimmune diseases and B cell malignancies.1 BAFF is a membrane-bound, type II transmembrane protein that belongs to the tumor necrosis factor (TNF) ligand family and is the ligand for BR3, TACI, and BCMA. BAFF is an immunostimulant necessary for maintaining normal immunity. This cytokine has also been shown to play an important role in the proliferation and differentiation of B cells. An inadequate level of BAFF leads to immunodeficiency whilst an elevated level of BAFF causes unusually high antibody production that results in the development of autoimmune diseases such as systemic lupus erythematosus and rheumatoid arthritis. Additionally, BAFF has been found in renal transplant biopsies with acute rejection.2 Furthermore, BAFF may be a mediator of food-related inflammation, and is associated with multiple dietary ailments including celiac disease, insulin resistance, diabetes, and obesity.3 Interestingly, it is suspected that BAFF may be involved in non-IgE-mediated reactions because there is no known correlation between BAFF and IgE.4 More research is needed to unlock the enormous therapeutic potential for BAFF antagonists. This cost-effective, research-grade Anti-Human CD257 (BAFF) (Tabalumab) utilizes the same variable regions from the therapeutic antibody Tabalumab making it ideal for research projects.
Antigen Distribution
BAFF is expressed on various cell types including monocytes, dendritic cells and bone marrow stromal cells.
Ligand/Receptor
TACI, BCMA,APRIL ligand, BAFFR/BR3
PubMed
NCBI Gene Bank ID
UniProt.org
Research Area
Biosimilars
.
Cancer
.
Cell Biology
.
Costimulatory Molecules
.
Immuno-Oncology
.
Immunology
.
Signal Transduction
.
Stem Cell

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 Tabalumab biosimilars are used as calibration standards or reference controls in pharmacokinetic (PK) bridging ELISA to enable quantitative measurement of drug concentration in serum samples.

The typical approach for biosimilar PK assays is to establish a single ligand-binding ELISA method using a single analytical standard—commonly, the biosimilar itself—as the calibrator for both biosimilar and reference product samples. This standard is used to generate a calibration curve in assay validation; serum samples containing either the biosimilar or reference Tabalumab are then quantified against this curve.

Key details on their use:

  • The biosimilar is rigorously characterized and validated for use as an analytical standard, ensuring that it matches the reference product in terms of assay accuracy and precision.
  • Validation includes preparation of biosimilar standards at defined concentrations in human serum (e.g., 50–12,800 ng/mL) and comparison of biosimilar and reference product recovery across QC samples over multiple assay runs by different analysts.
  • The use of a single biosimilar standard minimizes inter-assay variability and eliminates the need for multiple parallel methods, making it possible to robustly compare PK profiles between the biosimilar and the reference monoclonal antibody.
  • Analytical equivalence is verified by statistical analysis of accuracy and precision (bioanalytical comparability), using tight equivalence intervals, with calibration curves created using spiked biosimilar preparations.
  • Reference-grade biosimilars for Tabalumab can also be used as controls to verify assay specificity, performance, and to bridge measurements between the test biosimilar and reference molecule.
  • The same principle is employed for other therapeutic biologics measured by PK ELISA, where research biosimilars can serve as reference standards for assay calibration and method bridging.

In summary: In PK ELISA bridging studies for Tabalumab, research-grade biosimilars serve as the standard for calibration curves and QC controls, enabling accurate, precise quantification of drug concentrations in serum, and supporting regulatory comparability studies between biosimilar and reference products.

Standard flow cytometry protocols for validating expression levels or binding capacity of BAFF using a conjugated Tabalumab biosimilar (e.g., PE or APC-labeled) typically involve staining target cells with the labeled antibody and measuring fluorescence intensity to assess binding or surface expression. While explicit protocols for Tabalumab-PE/APC conjugates are not widely published, protocols for Tabalumab and related antibodies follow established principles in the field.

Key protocol steps include:

  • Cell Preparation:

    • Use cells engineered to express BAFF or cells naturally expressing BAFF (such as activated monocytes).
    • Detach adherent cells using enzyme-free dissociation (for cell integrity).
    • Wash cells and resuspend in FACS buffer (e.g., D-PBS, 2% FBS, 0.1% sodium azide).
  • Blocking Steps:

    • Add excess IgG (e.g., goat IgG at 1 mg/mL) to reduce non-specific Fc binding.
  • Staining:

    • Incubate cells with the labeled Tabalumab biosimilar antibody (e.g., PE-Tabalumab) at an optimized concentration, typically on ice or at 4°C for 20–30 minutes.
    • For indirect staining, pre-incubate BAFF-expressing cells with unlabeled Tabalumab, then follow with a fluorophore-labeled secondary antibody or streptavidin conjugate if using biotinylated BAFF.
    • Include an isotype control to establish specificity of binding.
    • Wash cells to remove unbound antibody.
  • Measurement:

    • Analyze by flow cytometer using appropriate excitation/emission settings for chosen fluorophore (e.g., PE, APC).
    • Gate on viable, single cells, and analyze fluorescence intensity to quantify BAFF expression or Tabalumab binding.

Essential controls:

  • Isotype control antibodies for background staining determination.
  • Unlabeled/unstained cells to establish baseline fluorescence.
  • Cells known to lack BAFF expression as negative controls.

Example protocol based on published BAFF–Tabalumab binding studies:

  1. Resuspend 1 million HEK293-BAFF or monocytes in FACS buffer.
  2. (Optional) Pre-incubate biotinylated BAFF with excess Tabalumab for competitive binding assessment, then add to cells.
  3. Incubate with fluorochrome-labeled Tabalumab biosimilar for 20 minutes on ice.
  4. Wash cells twice in FACS buffer.
  5. Resuspend in FACS buffer and proceed to data acquisition and analysis.

Additional notes:

  • Tabalumab was shown to specifically bind both soluble and membrane-bound BAFF, confirming suitability for flow cytometry-based target validation.
  • A biosimilar antibody with identical variable regions as Tabalumab, conjugated to PE or APC, will mimic Tabalumab binding profiles for flow-based BAFF quantification.

These steps align with standard practices for antibody-based flow cytometry analysis. Direct conjugation of Tabalumab biosimilar to PE or APC follows typical labeling protocols and does not alter fundamental workflow. Adapting for Tabalumab specificity ensures reliable BAFF detection on target cells.

Biopharma companies typically perform a comprehensive range of analytical assays to confirm both the structural and functional similarity of a proposed biosimilar to the originator (reference) biologic. These studies adhere to stringent regulatory guidelines, comparing the biosimilar and the original drug across multiple critical quality attributes (CQAs), using highly sensitive and orthogonal analytical methods.

Key Analytical Assays for Biosimilarity:

  • Structural Comparisons:

    • Primary Structure: Peptide mapping, mass spectrometry (MS), and amino acid sequencing to confirm the exact sequence of amino acids.
    • Higher-Order Structure: Circular dichroism (CD), nuclear magnetic resonance (NMR), X-ray crystallography, and hydrogen-deuterium exchange (HDX) MS for assessing protein folding (secondary, tertiary, quaternary structures).
    • Post-Translational Modifications: Glycan analysis, identification of disulfide bonds, and modification profiling using MS and chromatographic methods.
    • Charge Variants and Isoforms: Isoelectric focusing (IEF) and ion-exchange chromatography.
    • Aggregate/Impurity Analysis: Size-exclusion chromatography (SEC), analytical ultracentrifugation (AUC), and other impurity and aggregate assessment techniques.
  • Functional Comparisons:

    • Binding Assays: Surface plasmon resonance (SPR), enzyme-linked immunosorbent assay (ELISA), or flow cytometry to measure target binding.
    • Cell-Based Bioassays: To assess the biological activity (e.g., cytotoxicity, proliferation, receptor activation).
    • Potency Assays: Functional confirmation that the biosimilar mediates the same biological result as the reference (e.g., receptor binding, immune effector functions).
    • Fc Function (for antibodies): Fcγ receptor binding and complement activation assays, particularly to demonstrate that differences in glycosylation do not affect immune function.

A rigorous head-to-head comparison across multiple lots of both the biosimilar and the reference product is standard, providing detailed evidence that any detected differences are not clinically meaningful.

Leinco Biosimilars in Analytical Studies:

Leinco Technologies acts as a supplier of well-characterized biosimilar reference materials. In biosimilar development, Leinco biosimilar proteins can serve as either:

  • In-study positive controls: Where the analytical assays require a biosimilar version as a benchmark alongside the originator molecule to validate the assay system.
  • Reference standards: To help calibrate or standardize assay performance.

This facilitates:

  • The assessment of assay sensitivity and specificity.
  • The establishment of comparability thresholds.
  • Cross-laboratory consistency, especially when internal controls or additional comparator standards are needed for robust analytical or functional assessment.

However, Leinco biosimilars are not typically used as direct "subjects" of regulatory comparability; instead, they support assay development, validation, or serve as auxiliary comparators when designing or troubleshooting analytical workflows.

In summary, the analytical similarity exercise in biosimilar development is multifaceted, relying on structural, physicochemical, and functional analyses, often using commercial biosimilar proteins like those from Leinco as references or assay controls as part of this rigorous process.

References & Citations

1. Manetta, J. et al. (2014) J Inflamm Res. 7: 121–131
2. Clatworthy, MR. et al. (2013) Transplantation. 96(4): 413–420.
3. Lied, GA. and Berstad, A. (2011) Scand J Immunol. 73(1):1-7.
4. Büchler, JR. and Cano, MN. (1986) Jpn Heart J. 27(1):117-22.

Formats Available

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