Anti-Human CD257 (BAFF) (Tabalumab) – Biotin

Anti-Human CD257 (BAFF) (Tabalumab) – Biotin

Product No.: LT1401

- -
- -
Product No.LT1401
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
0.5 mg/ml
Formulation
This Biotinylated antibody 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 biotinylated antibody 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
Additional Reported Applications For Relevant Conjugates ?
N
IP
WB
IF
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 assays primarily to ensure quantitative and consistent measurement of Tabalumab concentrations in serum samples.

Key roles and procedures:

  • Calibration Standard Selection: The biosimilar is prepared as an analytical standard, meaning it serves as the reference against which unknown concentrations in serum samples are measured. The selection is based on validated bioanalytical comparability between the biosimilar and the reference product.

  • Standard Curve Generation: Multiple concentrations of the biosimilar standard are spiked into human serum to generate a standard curve, usually spanning the relevant quantification range for the assay (e.g., 50–12800 ng/mL in validated Tabalumab studies). This enables precise interpolation of drug levels in patient samples based on their ELISA signal.

  • Reference Controls: Quality control (QC) samples are prepared both with the biosimilar and, if available, reference product to monitor accuracy and precision throughout the assay runs.

  • Assay Validation: The method is validated for parameters such as accuracy, precision, selectivity, and sensitivity, using samples prepared with the biosimilar standard. Statistical analysis confirms that responses for biosimilar and originator Tabalumab are equivalent (bioanalytical comparability) within the established method.

  • PK Bridging Studies Use:

    • Serum samples from subjects administered Tabalumab are assayed using the biosimilar standard curve.
    • The measured concentrations are used for PK analysis and comparison across products.
    • Establishes equivalence and supports regulatory submissions for biosimilar approval.
  • Advantages of Single Standard Systems:

    • Reduces inter-assay variability.
    • Avoids the need for multiple methods or crossover calibration when comparing products in blinded clinical trials.

Summary Table: Use of Tabalumab Biosimilars in PK Bridging ELISA

ApplicationFunctionalityRationale
Calibration StandardGenerate standard curve for quantificationEnsures consistent, accurate measurement
Reference (Quality Control)Monitor assay accuracy/precision across runsValidates assay and supports equivalence
Assay validation and qualificationConfirm bioanalytical comparability, method suitabilitySatisfies regulatory/industry standards

Supporting Details:

  • Regulatory and industry guidelines favor a single PK assay using a single analytical standard for biosimilar and reference product quantification, provided rigorous comparability has been established.
  • Commercial suppliers offer biosimilars specifically for use as ELISA reference standards or as capture/detection reagents in bridging assays.

In summary, research-grade Tabalumab biosimilars are integral for reliable and standardized quantitative measurement in PK bridging ELISA assays, serving as calibration standards and reference controls once their equivalence with the originator is validated.

Standard flow cytometry protocols for validating the expression or binding capacity of the BAFF (B-cell Activating Factor) target with a conjugated Tabalumab biosimilar (e.g., PE or APC-labeled) generally involve staining BAFF-expressing cells with the labeled antibody and quantifying binding via fluorescence analysis. Below is a synthesis of standard approaches tailored to your scenario, referencing Tabalumab and best practices for antibody biosimilar validation:

Key Steps in Flow Cytometry Protocol:

  • Cell Preparation:

    • Use either cell lines transfected to express BAFF (e.g., HEK293-BAFF, BR3, TACI, BCMA) or primary human monocytes activated to upregulate BAFF.
    • Harvest cells gently (e.g., enzyme-free dissociation), resuspend at ~10^6 cells/mL in FACS buffer (DPBS + 2% FBS + 0.1% sodium azide), and block Fc receptors if necessary (by adding species-matched IgG).
  • Antibody Staining:

    • Incubate cells with the conjugated Tabalumab biosimilar (directly labeled with PE, APC, etc.) at a recommended concentration (typically empirically titrated), for 15–30 minutes at 4°C or on ice to minimize internalization and nonspecific binding.
    • If using an indirect method (biotinylated Tabalumab plus labeled streptavidin), incubate first with biotinylated antibody, wash, then incubate with fluorescent streptavidin.
  • Controls:

    • Include appropriate negative and positive controls, such as:
      • Cells with no antibody (autofluorescence)
      • Cells with isotype control antibody-fluorochrome conjugate
      • Known negative BAFF-expressing cells
      • For competitive binding assays, pre-incubate with excess unlabeled Tabalumab or BAFF
  • Washing:

    • Wash cells thoroughly with FACS buffer after each staining or incubation step to remove unbound antibody and reduce background.
  • Data Acquisition:

    • Analyze stained cells on a flow cytometer compatible with the chosen fluorochrome (e.g., PE or APC).
    • Collect sufficient events per sample (e.g., at least 2,000 cells as a minimum for rare populations as described in Tabalumab characterization studies).
  • Data Analysis:

    • Quantify fluorescence intensity to determine BAFF expression or binding capacity.
    • Overlay histograms or dot plots to compare staining between samples and controls.

Key Additional Notes:

  • Fc-Mutated Biosimilars: Many biosimilar antibodies used in flow cytometry have Fc mutations (e.g., N297A) to reduce Fc receptor–mediated background, increasing specificity and assay clarity.
  • Panel Compatibility: Choose fluorochromes that avoid spectral overlap with other panel components if multiplexing.
  • Assay Validation: Ensure validation for specificity, repeatability, and precision per standard bioassay guidelines (see general principles from validation studies on biosimilar flow cytometry assays).
  • Application Example: In the original characterization of Tabalumab, binding to membrane-bound BAFF was validated by flow cytometry using either direct or indirect fluorescence labeling, allowing assessment of both binding capacity and expression levels in engineered and primary human cells.

This protocol framework supports rigorous validation of the binding and specificity of a PE/APC-labeled Tabalumab biosimilar to its BAFF target in either overexpression cell lines or primary cells, and can be adapted to suit your specific research or development setting.

Biopharma companies typically perform a comprehensive set of analytical assays to establish both structural and functional similarity between a proposed biosimilar and its reference (originator) drug, focusing on critical quality attributes (CQAs) and using orthogonal approaches for rigorous comparison.

Structural assays commonly performed include:

  • Primary structure analysis: Peptide mapping by mass spectrometry for confirming the amino acid sequence and detecting variations like deamidation or oxidation.
  • Protein heterogeneity: Assessing charge variants and aggregates using UHPLC or capillary electrophoresis.
  • Glycosylation analysis: Advanced workflows to characterize glycoforms, released glycans, glycopeptides, and monosaccharides, often including sialic acid content.
  • Secondary, tertiary, and quaternary structure: Circular dichroism (CD), differential scanning calorimetry (DSC), and analytical ultracentrifugation.
  • Impurity profile and purity: Comparing aggregates, fragments, and other product-related variants.

Functional assays typically include:

  • Biological activity assays: Cell-based assays tailored to the drug’s mechanism of action.
  • Binding assays: ELISA or surface plasmon resonance (SPR) to compare antigen or receptor binding affinity.
  • Potency assays: Measuring functional response such as receptor activation, effector function, or enzyme activity.
  • Fc effector function (for antibodies): ADCC, CDC, or FcγR binding assays, especially if small glycosylation differences are observed.

These assays are usually performed on multiple lots of both the biosimilar and reference product, with data compared to ensure all attributes fall within a predefined similarity range.

Regarding Leinco biosimilars:
Leinco Technologies specializes in supplying highly characterized biosimilar antibodies and proteins for research, analytical assay development, and product comparability studies. Leinco biosimilars can serve as:

  • Analytical controls or comparators to validate assay accuracy and specificity, especially when developing head-to-head comparison workflows.
  • Reference materials for benchmarking in structural fingerprinting and biological activity testing, ensuring that analytical methods appropriately distinguish and quantify critical quality attributes.

Leinco biosimilars are thus used in these studies predominantly as trusted reagents for assay development, method validation, and sometimes as surrogate standards when the branded reference product is unavailable or in limited supply. Specific details of use depend on the study protocol and regulatory requirements at each stage of biosimilar development.

In summary, the confirmation of biosimilarity relies on an array of structural and functional assays, many of which utilize biosimilar reagents—such as those from Leinco—for robust, reproducible, and regulatory-compliant analyses.

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.