Challenges with development of a pharmacokinetics assay to measure a variably glycosylated fusion protein

G Sperinde, L Liu, K Xu et al. | Bioanalysis, 14(1), 7 - 18 (2021) Keywords: • assay • PK • sialic acid • variants Aim: Development of recombinant fusion proteins as drugs poses unique challenges for bioanalysis. This paper describes a case study of a glycosylated fusion protein, where variable glycosylation, matrix interference and high sensitivity needs posed unique challenges. Results: Six different assay configurations, across four different platforms were evaluated for measurement of drug concentrations. Two platforms that achieved the assay requirements were Simoa HD-1 and immune-capture LC–MS/MS-based assay. Conclusion: Both, Simoa HD-1 and the mass spectrometry-based methods...