Antibody System Anti-Vosoritide Neutralizing Antibody ELISA Kit
Overview
This enzyme-linked immunosorbent assay (ELISA) kit is engineered for the precise quantitative determination of neutralizing antibodies against Vosoritide in serum and plasma samples. Vosoritide is a modified recombinant human C-type natriuretic peptide (CNP) analog comprised of 39 amino acids, featuring an extended N-terminus that prolongs its half-life by resisting neutral endopeptidase degradation. Administered primarily to treat achondroplasia—the most common form of human dwarfism—Vosoritide acts as a targeted therapeutic agent by binding specifically to Natriuretic Peptide Receptor B (NPR-B). This binding cascade inhibits the overactive Fibroblast Growth Factor Receptor 3 (FGFR3) signaling pathway, which downstream promotes endochondral bone growth and chondrocyte proliferation. However, chronic administration can prompt an immune response leading to the generation of anti-drug antibodies (ADAs). Neutralizing antibodies represent a critical subset of ADAs that explicitly bind to the receptor-interaction domains of Vosoritide, physically blocking it from engaging the endogenous NPR-B receptor. This competitive inhibition compromises the drug's therapeutic efficacy. This competitive ELISA serves as a vital analytical instrument for clinical investigators, geneticists, and biomanufacturers profiling long-term immunogenicity, monitoring therapeutic resistance, and validating biosimilar discovery pipelines.
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:
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Target Capture: Recombinant Vosoritide is uniformly pre-coated onto the solid-phase surface of a 96-well microplate.
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Competitive Binding: Calibrated reference standards or unknown biological samples are premixed with a biotin-labeled NPR-B receptor tracer and pipetted directly into the microwells. Endogenous Anti-Vosoritide Neutralizing Antibodies present within the sample compete directly with the biotinylated receptor tracer for a limited number of immobilized Vosoritide binding domains.
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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 receptor domains.
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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 receptor tracer binding, the resulting color intensity develops in inverse proportion to the concentration of active Anti-Vosoritide 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
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Sensitive Detection Capability: Exhibits an analytical sensitivity down to 134.11 ng/mL, allowing investigators to track low-affinity or early-stage neutralizing antibody formations.
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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.
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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.
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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
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Catalog No.: KAD59502
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Assay Configuration: Quantitative Competitive Enzyme Immunoassay
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Targeted Biomarker: Anti-Vosoritide Neutralizing Antibodies
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Validated Matrices: Serum and Plasma
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Assay Range: 156.25 to 10,000 ng/mL
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Analytical Sensitivity: 134.11 ng/mL
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Solid-Phase Target: Recombinant Vosoritide
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Detection System: Biotin-NPR-B / 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:
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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.
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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
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Kit Shipping: Distributed under secure, temperature-controlled conditions on blue ice or chilled gel packs to preserve protein matrix and enzyme binding configurations.
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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.
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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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