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Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery

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Mitigating target interference in bridging immunogenicity assay with target-blocking reagents and mild basic pH.

Jihua Chen¹, John Garlits¹, Gangadhar Dhulipala¹

¹Regeneron Pharmaceuticals, Inc., 777 Old Saw Mill River Road, Tarrytown, NY 10591, USA.

|June 19, 2019

Summary

False positive results in anti-drug antibody (ADA) assays were reduced using novel blocking strategies. These methods improve assay accuracy by mitigating target interference, ensuring reliable detection of therapeutic antibodies.

Keywords:

anti-drug antibody immunogenicity mild basic pH monoclonal antibody therapeutic target interference

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Area of Science:

Biochemistry
Immunology
Assay Development

Background:

Soluble drug targets in clinical samples cause false positives in anti-drug antibody (ADA) bridging assays via target-mediated bridging.
This interference complicates the accurate assessment of drug immunogenicity.

Purpose of the Study:

To develop and validate strategies to mitigate target interference in ADA bridging assays.
To improve the reliability and accuracy of ADA detection in clinical samples.

Main Methods:

Utilized a combination of two target-blocking reagents.
Optimized assay conditions by adjusting to a mild basic pH.
Tested assay performance with known ADA-positive and negative samples, including those with specific pharmacokinetic profiles.

Main Results:

The combined strategy demonstrated high tolerance to recombinant target protein.
Reduced false positive rates were observed in clinical study samples.
The assay successfully detected ADA-positive samples from nonclinical studies with minimal impact from the modified conditions.

Conclusions:

The developed strategies offer effective alternatives for mitigating target interference in ADA assays.
These methods are particularly useful when standard target-blocking antibodies are insufficient.
Improved assay robustness enhances the assessment of drug immunogenicity.