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Development and Validation of Cell-Based Bioassay for the Detection of Neutralizing Antibodies to Ocrelizumab in
Olga Strizhakova1, Grigory Poroshin1, Andrei Pershin1
1JSC "GENERIUM", 14, Vladimirskaya Street, Volginskiy 601125, Vladimir Region, Russia.
Background/Objectives:
Ocrelizumab is a humanized monoclonal antibody targeting CD20, approved for the treatment of adult patients with relapsing multiple sclerosis (RMS) and primary progressive multiple sclerosis (PPMS). The neutralizing activity of anti-drug antibodies (ADAs), especially neutralizing ADAs (nADAs) activity, should be examined considering that it can alter pharmacokinetic (PK) and pharmacodynamic (PD) profiles, reduce drug efficacy, and lead to life-threatening adverse events.
Methods:
This article presents data on the development and validation of an assay for neutralizing anti-drug antibodies (nADA) based on ADCC reporter cells for the analysis of patient sera in the context of ocrelizumab clinical studies.
Results:
Critical steps and conditions to minimize assay variability were identified. The lower limit of detection was 549.6 ng/mL. The cutoff for nonspecific neutralization was determined as 19.7%. The presence of 0.37-3.0 μg/mL ocrelizumab in a biological sample enables the detection of 1.1-10.0 μg/mL polyclonal anti-ocrelizumab idiotype antibodies, respectively.
Conclusions:
The developed method can be used for immunogenicity studies of medicinal products containing ocrelizumab.
Insights
A new assay was developed to detect neutralizing anti-drug antibodies (nADAs) against ocrelizumab. This method is crucial for monitoring drug efficacy and safety in patients with multiple sclerosis (MS).
Area of Science:
- Immunology
- Pharmacology
- Biotechnology
Background:
- Ocrelizumab, a CD20-targeting monoclonal antibody, treats relapsing and primary progressive multiple sclerosis (MS).
- Anti-drug antibodies (ADAs), particularly neutralizing ADAs (nADAs), can impact ocrelizumab's pharmacokinetic/pharmacodynamic profiles, efficacy, and safety.
- Assessing nADA activity is vital for ocrelizumab's clinical use.
Purpose of the Study:
- To develop and validate a robust assay for detecting neutralizing anti-drug antibodies (nADAs) against ocrelizumab.
- To support immunogenicity assessments in ocrelizumab clinical studies.
Main Methods:
- Development of a neutralizing anti-drug antibody (nADA) assay utilizing ADCC reporter cells.
- Validation of the assay for analyzing patient sera in ocrelizumab clinical trials.
- Identification of critical steps to minimize assay variability.
Main Results:
- The assay demonstrated a lower limit of detection of 549.6 ng/mL.
- A cutoff for nonspecific neutralization was established at 19.7%.
- The assay can detect polyclonal anti-ocrelizumab idiotype antibodies within specific concentration ranges when ocrelizumab is present.
Conclusions:
- The developed assay is suitable for immunogenicity studies of ocrelizumab-containing medicinal products.
- This method aids in the comprehensive evaluation of ocrelizumab's therapeutic use.
