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Antibody System Anti-Ocrelizumab Neutralizing Antibody ELISA Kit

Antibody System Anti-Ocrelizumab Neutralizing Antibody ELISA Kit

Overview
This enzyme-linked immunosorbent assay (ELISA) kit is engineered for the precise quantitative determination of neutralizing antibodies against Ocrelizumab in serum and plasma samples. Ocrelizumab is a humanized monoclonal IgG1 antibody that binds selectively to the CD20 antigen, a hydrophobic transmembrane protein primarily expressed on the surface of pre-B and mature B lymphocytes. By targeting CD20, Ocrelizumab mediates B-cell depletion through antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC), serving as a paramount disease-modifying therapy for relapsing and primary progressive forms of multiple sclerosis (MS). While clinically highly effective, chronic administration can provoke a humoral immune response leading to the generation of anti-drug antibodies (ADAs). Neutralizing antibodies represent a critical subset of ADAs that explicitly bind to the antigen-binding domains (CDR loops) of Ocrelizumab, physically blocking it from engaging the endogenous human CD20 receptor. This specific inhibition compromises B-cell depletion kinetics, driving therapeutic non-responsiveness and disease progression. This competitive ELISA serves as a vital analytical instrument for clinical investigators, neurologists, and biomanufacturers profiling long-term immunogenicity, monitoring therapeutic resistance, and validating biosimilar discovery pipelines.
 
DATASHEET

  • PRODUCT INFO

    Principle of the Assay

    This assay utilizes a high-performance, solid-phase quantitative competitive enzyme immunoassay technique to measure neutralizing anti-drug antibody levels:

    • Target Capture: Recombinant Ocrelizumab is uniformly pre-coated onto the solid-phase surface of a 96-well microplate.

    • Competitive Binding: Calibrated reference standards or unknown biological samples are premixed with a biotin-labeled Human CD20 antigen tracer and pipetted directly into the microwells. Endogenous Anti-Ocrelizumab Neutralizing Antibodies present within the sample compete directly with the biotinylated CD20 tracer for a limited number of immobilized Ocrelizumab binding domains.

    • Enzyme Conjugation: Unbound matrix elements and non-neutralizing fractions are cleared via a rigorous wash cycle. Streptavidin conjugated to Horseradish Peroxidase (Streptavidin-HRP) is introduced into the wells, anchoring specifically to the remaining captured biotin-labeled antigen domains.

    • Signal Amplification and Readout: Following a final wash to remove excess enzyme reagent, a colorimetric substrate solution is added to initiate enzymatic signal development. Because this is a competitive assay format where neutralizing antibodies physically prevent antigen tracer binding, the resulting color intensity develops in inverse proportion to the concentration of active Anti-Ocrelizumab Neutralizing Antibodies bound in the initial step. The reaction is terminated with a stop solution, and absorbance is quantified spectrophotometrically.

    Key Features and Performance Metrics

    • Sensitive Detection Capability: Exhibits an analytical sensitivity down to 132.56 ng/mL, allowing investigators to track low-affinity or early-stage neutralizing antibody formations.

    • Extended Quantitative Range: Delivers a reliable linear tracking range from 156.25 ng/mL to 10,000 ng/mL to seamlessly accommodate diverse sample concentration profiles without immediate saturation.

    • Consistent Intra-Assay Precision: Demonstrates reliable target reproducibility with a Coefficient of Variation (CV) of less than 20 percent, validated through sixteen separate replicate evaluations across three samples of known concentration on a single test plate.

    • Outstanding Inter-Assay Uniformity: Confirms high day-to-day and batch-to-batch experimental reliability with a CV of less than 20 percent, verified across twenty-four distinct assay runs using three independent reference concentration samples.

    Technical Specifications

    • Catalog No.: KAC90704

    • Assay Configuration: Quantitative Competitive Enzyme Immunoassay

    • Targeted Biomarker: Anti-Ocrelizumab Neutralizing Antibodies

    • Validated Matrices: Serum and Plasma

    • Assay Range: 156.25 to 10,000 ng/mL

    • Analytical Sensitivity: 132.56 ng/mL

    • Solid-Phase Target: Recombinant Ocrelizumab

    • Detection System: Biotin-Human CD20 / Streptavidin-HRP Conjugate

    Applications and Workflow Summary

    This competitive ELISA platform is highly optimized for therapeutic drug immunogenicity monitoring, biosimilar validation, and downstream pharmacokinetic profiling. Investigators should observe the following guidelines:

    • Dilution and Layout: Unknown serum or plasma matrices must be screened alongside the provided reference standards. Samples exhibiting target concentrations exceeding the maximum range of 10,000 ng/mL must be diluted empirically in standard assay matrix buffer prior to running the assay.

    • Analysis Requirements: To achieve absolute quantification accuracy, investigators must generate a standard curve simultaneously during every plate run, plotting standard absorbance profiles against log-transformed concentration values.

    Handling, Stability and Storage

    • Kit Shipping: Distributed under secure, temperature-controlled conditions on blue ice or chilled gel packs to preserve protein matrix and enzyme binding configurations.

    • Operational Shelf Life: When the kit is stored continuously at the recommended temperature for 6 months, the internal signal intensity and enzymatic reactivity profile degrade by less than 20 percent.

    • Storage Guidance: Keep the complete kit stored strictly at the manufacturer-specified refrigeration temperature upon receipt. Ensure that unused microplate wells remain sealed within their original desiccant pouch to guard against moisture compromise.

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