Oligonucleotide (OGN) therapy is increasingly recognized as an important therapeutic modality with the promise of curing diseases. However, unlike most biotherapeutics, the development of OGN drugs often requires their determination with high sensitivity and specificity in multiple, complex biologic matrices such as blood and tissues in order to measure their exposure and distribution for understating PK, PD, and safety.
At Frontage Laboratories (PA, USA), we have developed highly sensitive and specific quantitative methods for the determination of OGN in plasma and tissues using hybridization immunoassay and LC–MS techniques. In this webinar, we will present the method development, validation, and comparison of these techniques for the quantification of OGN as well as a case study with antisense OGN.
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Antisense oligonucleotides (ASOs) are short, chemically modified, single-stranded DNA/RNA oligonucleotides that specifically target genes of interest and regulate target protein biosynthesis. In this study, we quantify antisense oligonucleotides in plasma using MSD.
Fascin is an actin-bundling protein that has been linked to tumor cell migration, invasion, metastasis, disease progression and mortality, thus serving as a novel cancer biomarker. In this paper, we develop and validate an LC-MS/MS method for the quantification of fascin proteins in human serum.
XMT-1522, an antibody-drug conjugate (ADC) currently in Phase I clinical development, represents the first Dolaflexin®-based, cleavable ADC with a high drug-antibody ratio (DAR). In this work, a novel immunocapture LC–MS/MS method was successfully developed for the simultaneous quantification of both total antibody and cleavable antibody-conjugated drug auristatin F-hydroxypropylamide (AF-HPA) in human plasma. This method utilized microwave-assisted enzymatic digestion for the total antibody and chemical release of the drug from ADC on a 96-well based immunocapture sample preparation platform. The total antibody and the conjugated drug AF-HPA were separated and subsequently quantified concurrently by LC–MS/MS. The linear range of the standard curve for total antibody was from 50 to 5000 ng/mL and for AF-HPA was from 3.3 to 330 ng/mL. The linearities showed R2 ≥ 0.993 for total antibody and R2 ≥ 0.996 for AF-HPA, respectively. The intra- and inter-day precision and accuracy were well within 15%. The validated method, with the characteristics of high efficiency, great selectivity, free of carryover, short LC–MS/MS time (˜3.5 min) and low sample volume (20 μl), was successfully applied for analyzing Phase 1 cancer patient samples.