Fit-for-purpose protein biomarker assay validation strategies using hybrid immunocapture-liquid chromatography-tandem-mass spectrometry platform: Quantitative analysis of total soluble cluster of differentiation 73.

摘要: Abstract In recent years, biomarkers have played more extensive roles as indicators of disease progression, safety, and drug efficacy. Targeted quantitative analysis including targets become increasingly important to drive critical decision-making in various development stages, well improve the success rates clinical trials. There are many analytical challenges when developing validating bioanalytical methods associated with measurement an endogenous protein biomarker, especially using LC-MS based analysis. Moreover, current regulatory guidelines for assay validation platform mainly focuses on regulated bioanalysis therapeutic drugs. this manuscript, we use total soluble CD73 (sCD73) example present a “fit-for-purpose” hybrid immunocapture-LC-MS/MS platform. A non-competing antibody (to drug) was used isolate enrich sCD73 from biological matrix. The enriched sample digested after immunocapture surrogate peptide monitored quantification. showed good accuracy, precision, specificity sensitivity LLOQ 1.00 ng/mL, applied study measure potential pharmacodynamic (PD) marker. Some recommendations considerations assay, assays general, is also discussed.