(T1130-01-04) Validation of an Anti-drug Antibody (ADA) Assay Using the Mitra VAMS Microsampling Technology in Support of a Phase I Clinical Study

Purpose: Detection of immune responses against a biological therapeutic is necessary and required by regulatory authorities in the context of their impact in safety and efficacy. Historically, liquid serum samples were used for anti-drug antibody (ADA) detection. Microsampling technologies allow collection and testing of low volume blood samples (especially important for pediatric population) and enable at-home sample collection. This study explored use of Mitra VAMS microsampling technology in an ADA assay to support a monoclonal therapeutic phase I clinical trial.

Methods: Bridging ADA assays with acid dissociation were validated for human serum and dried human blood sample matrices. Mitra VAMS tips were extracted in PBST prior to analysis. Anti-drug antibodies extracted from dried whole blood maintained their fidelity and capacity for binding drug molecules. Tecan automation was implemented to prepare large numbers of VAMS negative and positive controls to support the clinical program.

Results: Both assays have comparable sensitivity of below 10 ng/mL, and drug tolerance above the expected C_trough level and can detect 100 ng/mL of ADAs in the presence of over 400 µg/mL of the therapeutic drug. 1,137 human serum samples and 908 dried blood samples from phase I study were analyzed. 18 (1.6%) human serum samples and 10 (1.1%) dried blood samples were confirmed positive for ADA. Of the 15 samples from the same subjects/time points from serum and dried blood, 9 samples were confirmed positive in both matrices, 5 confirmed positive serum samples had dried blood counterparts that were negative, 1 confirmed positive dried blood sample had serum counterpart that was negative

Conclusion: Our results suggest that the Mitra VAMS microsampling technology is technically compatible with traditional serum sampling in bridging ADA assays following acid disassociation. Implementation of such technologies has the potential to reduce sample volume requirement and enable at-home sampling for long term immunogenicity monitoring.


Validation Summary Table


Clinical Results Graph 1


Clinical Results Table 1

Acknowledgements: Thomas Steinmetz M.S. for providing statistical analysis and support.