GEN-MKT-18-7897-A
Oct 10, 2016 | Biopharma, Blogs | 0 comments
By the End of 2024, the peptide therapeutics market value is expected to reach US$46.6 billion1. However, peptide therapeutics present some of the toughest analytical challenges for bioanalytical quantification. Large molecule therapeutics often may not fragment well for MS/MS analysis and often the case upon digestion in a complex biologic matrix this leads to an even higher background and potential for interferences.
To keep up with the pace of developmental therapies, Novartis Pharmaceuticals, along with SCIEX application specialists, recently developed a validated method to quantify pasireotide, a therapeutic peptide used to treat Cushing’s disease, in human plasma samples. The complete article was published in Analytical Chemistry and highlights how SelexION Technology on a QTRAP system can be used to increase selectivity for the accurate and sensitive quantitation of the difficult peptide therapeutic. To summarize the article, pasireotide, a therapeutic cyclic peptide exhibits poor fragmentation efficiency for multiple reaction monitoring (MRM) detections. Therefore, to increase the overall sensitivity of the assay, a DMS multiple ion monitoring (MIM) approach was explored. This method, utilizing the SelexION DMS cell on a QTRAP 6500, overcomes the sensitivity challenge in the typical MRM method due to the poor fragmentation of the analyte. What is great about this is the Pasireotide Method was developed and validated for bioanalytical analysis in human plasma down to 0.01 ng/mL, approximately 5-fold better than an MRM based method without using DMS separation – talk about improvements!
Over the past few years, researchers have needed to overcome the challenges associated with peptide bioanalysis in order to keep up with market demands. One way to do this according to Evgueni Fedorov, Executive Director, R&D, BIOTRIAL Bioanalytical ServicesOne, is to increase separation power. This was demonstrated in his webinar with SCIEX, where he showed how SelexION DMS technology on a QTRAP system can be used to enhance the selectivity of another challenging large therapeutic peptide, exenatide.
The Take-Away
References
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As an analytical strategy, middle-down mass spectrometry (MS) workflows characterize biotherapeutic proteins by analyzing large, digested protein fragments or defined subunits, rather than fully intact proteins (top-down) or digested peptides (bottom-up). A middle-down strategy combines the strengths of top-down and bottom-up approaches by delivering high sequence coverage and structural specificity while maintaining relatively simple sample preparation. In practice, middle-down analysis enables accurate mass measurement, rapid sequence confirmation, and localization of key post-translational modifications (PTMs) on protein subunits that are directly relevant to product quality.
In biopharmaceutical development, sequence variants (SV) are considered an inherent risk of producing complex proteins in living systems. Sequence variants are unintended changes to the amino acid sequence of a biotherapeutic and can be caused by errors in transcription or translation in the host cell, or cell culture and process conditions. Detailed analysis of SVs is important in process and product development to ensure the drug’s safety and efficacy. Even low‑level sequence variants can have significant implications for product quality, safety, and efficacy, making their accurate detection and characterization a critical requirement across development, process optimization, and regulatory submission.
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