Read "An immunogenicity analysis strategy to reduce the interference of soluble target proteins"
Compared with traditional small molecule drugs, therapeutic proteins carry a higher risk of immunogenicity, which can reduce exposure, decrease efficacy, or even trigger adverse events. Accurate anti-drug antibody (ADA) assays are essential for evaluating pharmacodynamics and safety.
One major challenge in ADA testing is that soluble target proteins in patient samples can interfere with the assay, producing false positives that misrepresent immunogenicity risk.
At Crystal Bio Solutions, our experts established a blocking antibody strategy to overcome this challenge and validated it on the MSD platform.
By treating acid-dissociated samples with a specific antibody that blocks affinity capture, we were able to remove soluble target interference and reduce false positive signals. This approach provides a more reliable basis for immunogenicity evaluation in therapeutic protein development.
During ADA assays, soluble target proteins circulating in serum can bind in ways that mimic ADA signals, leading to false positives. This not only complicates data interpretation but may also misguide decisions on clinical pharmacodynamics, efficacy, and safety.
To address this, our experts integrated a blocking antibody step into an ACE (affinity capture elution) ADA assay workflow:
Acid dissociation to release ADA from complexes
Blocking antibody treatment to neutralize soluble targets before capture
MSD-based detection to quantify ADA accurately without interference
This strategy effectively distinguished true ADA signals from target-related artifacts.
The validated workflow showed:
Strong drug tolerance across multiple concentrations (100–500 µg/mL) with positive ADA detection at high positive control levels
Clear reduction of false positives in samples with elevated soluble target concentrations when blocking antibody was applied
Consistent results across multiple serum samples, confirming robustness of the approach
For therapeutic proteins, immunogenicity assessment is critical to ensure patient safety and clinical efficacy. False positives from soluble target interference can lead to overestimation of immunogenicity risk, delaying development or complicating regulatory submissions. By reducing this interference, our strategy ensures ADA assays produce accurate, reliable data, helping biopharma teams move forward with confidence.
Key Takeaways
Developed a blocking antibody strategy to mitigate soluble target interference in ADA assays
Successfully applied within an ACE assay on the MSD platform
Validated with robust drug tolerance and reduced false positives across serum samples
Provides a more accurate and reliable approach for immunogenicity assessment in therapeutic protein development