A validated approach for accurate PK measurement when antibodies fall short.
When traditional ligand-binding assays couldn’t deliver the specificity required for insulin aspart, Inotiv’s LC-MS Centers of Excellence developed a hybrid immunocapture/LC-MS/MS method that overcame antibody cross-reactivity and enabled confident PK decision-making.
Using a biotinylated anti-human insulin antibody bound to streptavidin-coated magnetic beads, the team isolated insulin aspart and a stable-isotope-labeled internal standard from human serum. The analytes were eluted and quantified using a Sciex 6500 LC-MS/MS system with a BEH C8 column and a rapid 4.12 minute cycle time.
This validated method demonstrated strong performance across accuracy, precision, recovery, matrix effects, and long-term stability—supporting 3,430 project samples across 60 runs in just four weeks. The same strategy successfully extended to insulin glargine and its M1/M2 metabolites, confirming broad applicability for insulin analog programs.
If your biosimilar, PK/PD, or biomarker program is facing antibody specificity challenges, our LC-MS Centers of Excellence can help you design, validate, and execute a fit-for-purpose hybrid assay.
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