Antibody System Anti-Alirocumab ELISA Kit

Principle of the Assay

This assay utilizes a high-performance, solid-phase quantitative sandwich enzyme immunoassay technique (bridging format) to measure specific anti-drug antibody levels:

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

• Specific Binding: Calibrated standards or unknown biological samples are pipetted directly into the microwells. Endogenous Anti-Alirocumab antibodies present in the sample are bound and captured by the immobilized Alirocumab proteins.

• Bridging Conjugation: Unbound matrix elements are removed via a rigorous wash cycle. A biotin-labeled Alirocumab tracer solution is introduced into the wells, binding to the open antigen-binding sites of the captured Anti-Alirocumab target, effectively forming an immune bridging complex.

• Enzyme Complexing: Following another wash step to clear unbound tracer, Streptavidin conjugated to Horseradish Peroxidase (Streptavidin-HRP) is added to bind specifically to the biotinylated domains.

• Signal Amplification and Readout: After a final wash to remove excess enzyme reagent, a colorimetric substrate solution is added to initiate enzymatic signal development. Because this is a bridging sandwich assembly, the resulting color intensity develops in direct proportion to the concentration of Anti-Alirocumab 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 101.12 ng/mL, facilitating the early tracking of low-level anti-drug antibody formations.

• Extended Quantitative Range: Delivers a wide dynamic tracking range from 156.25 ng/mL to 10,000 ng/mL to accommodate diverse sample concentration tiers without immediate saturation.

• Exceptional Assay Precision: Demonstrates excellent target reproducibility with a Coefficient of Variation (CV) of less than 15 percent, ensuring robust experimental uniformity.

• Reliable Recovery Profile: Validated with a target recovery range between 80 percent and 120 percent, indicating low matrix interference in both human plasma and serum testing setups.

Technical Specifications

• Catalog No.: KAJ24001

• Assay Configuration: Quantitative Bridging Sandwich Enzyme Immunoassay

• Targeted Biomarker: Anti-Alirocumab (Anti-Drug Antibodies / ADA)

• Validated Matrices: Plasma and Serum

• Assay Range: 156.25 to 10,000 ng/mL

• Analytical Sensitivity: 101.12 ng/mL

• Detection Method: Colorimetric

• Solid-Phase Target: Recombinant Alirocumab

• Detection Enzyme Layer: Biotin-Alirocumab / Streptavidin-HRP Conjugate

Alternative Names: Anti-REGN727, Anti-SAR-236553, Anti-SAR236553, Anti-SAR 236553, Anti-CAS 1245916-14-6.

Applications and Workflow Summary

This bridging sandwich ELISA platform is highly optimized for therapeutic drug immunogenicity monitoring 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 between 2 and 8 degrees Celsius to safeguard protein matrix and enzyme binding configurations.

• Operational Shelf Life: Built with high-grade components to guarantee consistent, low-decay signal retention, demonstrating an activity loss rate of less than 20 percent prior to the expiration date when kept under appropriate storage conditions.

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