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Biolayer Interferometry-based FcγRIIa binding assay for a therapeutic antibody with strong effector function
Adriana Bajardi-Taccioli1, Carl Co1, Christine Bond1
1Department of Analytical Development, Biogen, Inc., 225 Binney St., Cambridge, MA, 02142, USA.
Analytical Biochemistry
|August 7, 2020
Summary
Aggregates in therapeutic antibody samples can falsely elevate FcγRIIa binding potency. A novel Biolayer Interferometry (BLI) assay was developed to accurately measure FcγRIIa receptor interactions, overcoming aggregate interference.
Area of Science:
- Biochemistry
- Immunology
- Pharmaceutical Science
Background:
- FcγRIIa receptor binding is crucial for therapeutic antibody efficacy.
- Existing assays like AlphaScreen can be confounded by sample aggregates, leading to inaccurate potency measurements.
- Aggregate interference can mask critical changes in antibody binding affinity and effector function.
Purpose of the Study:
- To develop and qualify a novel assay for accurate FcγRIIa receptor binding assessment.
- To overcome the limitations of aggregate interference in FcγRIIa binding assays.
- To provide a robust method for therapeutic antibody characterization and quality control.
Main Methods:
- Development of a novel Biolayer Interferometry (BLI)-based FcγRIIa binding assay.
- Utilizing association binding responses and Parallel Line Analysis for potency calculation.
- Qualification of the BLI assay for reliability and accuracy.
Main Results:
- The novel BLI assay accurately measures FcγRIIa binding potency, unaffected by sample aggregates.
- The assay successfully distinguished between different Fc glycosylation patterns.
- The developed method proved to be stability-indicating.
Conclusions:
- A qualified BLI-based FcγRIIa binding assay effectively resolves aggregate interference issues.
- This assay is suitable for sample characterization, drug product release, and stability testing.
- The method enhances the reliability of therapeutic antibody assessment.
