GEN-MKT-18-7897-A
Jul 14, 2023 | Blogs, LC, Pharma, SCIEX OS software, ZenoTOF 7600 system | 0 comments
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So, you need a new liquid chromatography-mass spectrometry (LC-MS) system for your metabolite identification (metID) studies, and you are not sure which option is right for you. This blog provides an overview of the metID solutions offered by SCIEX, so you can make the best decision for your organization.
This is the first in a series of blogs intended to help you choose a system that meets your pharmaceutical research and development needs. We hope you find their guidance useful.
Let’s start with a simple question: Which of the following would help you sleep well at night?
We have great news for you: SCIEX has solutions for enabling both A and B. Let’s take a closer look.
Scenario A Let’s say you are looking for a system that will run larger batches of samples, and your analytes typically fragment well using collision-induced dissociation (CID), allowing confident identification of the site of metabolism without the need for additional confirmatory testing.
For a complex analysis, the requirements are relatively straightforward, and the priorities are probably ease of use and a robust system that does the job day in and day out. For this scenario, SCIEX recommends using the ExionLC 2.0+ system coupled to the X500R QTOF system, SCIEX OS software and maintenance with SCIEX service and support. The workflow described in the technical note Targeted high-resolution metabolite screening workflows demonstrates the performance you can expect from this solution.
Scenario B Suppose you have several samples where clear identification of the site of metabolism is not possible using CID. In this case, the use of a secondary analytical technique, such as nuclear magnetic resonance (NMR), is often required to meet regulatory requirements. Since this approach can be both time-consuming and expensive, an LC-MS system with more analytical power is ideal. Potent therapeutics is another common challenge in this type of scenario, which makes method sensitivity a key requirement for detecting low-abundant metabolites.
In these circumstances, SCIEX recommends using the ExionLC 2.0+ system coupled to the ZenoTOF 7600 system, SCIEX OS software and maintenance with SCIEX service and support.
The ZenoTOF 7600 system features the Zeno trap for enhanced sensitivity and electron activated dissociation (EAD) as a complementary fragmentation option. EAD typically fragments the metabolite in different locations, which can provide the additional information required for clear identification of the site of metabolism from a single analytical technique.
The workflow described in the technical note Confident characterization and identification of glucuronide metabolites using diagnostic fragments from electron activated dissociation (EAD) is a useful example of this solution.
Learn more For more information on the options SCIEX offers for metID, please speak to your account manager or visit our web page on comprehensive metID.
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.
CE‑SDS remains a cornerstone assay for characterizing fragmentation, aggregation, and product‑related impurities in therapeutic proteins. UV detection has been the long‑standing standard. However, it frequently struggles with baseline noise, limited sensitivity for minor fragments, and subjective integration.
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