GEN-MKT-18-7897-A
May 22, 2017 | Biopharma, Blogs | 0 comments
From small ions like phosphate, herbicide degradation to metabolites, oligosaccharides, peptides, and proteins. How is your lab analyzing polar molecules? The reason I ask is there is a saying, if you have a charged or polar molecule, look to capillary electrophoresis (CE) first. While liquid chromatography (LC) is an ideal front-end separation tool for many types of molecules, as the following poster points out, “From Small to Very Large, Orthogonal, Sensitive Polar Molecule Analysis by CESI-MS,” there are some situations that call for CE over LC analysis. For those of you that are not familiar with CESI-MS, it is the combining of CE separation with electrospray ionization, into one dynamic process, within the same device. View the Poster >
Complex Polar MoleculesLet me explain. Although effective, analysis by traditional LC-MS methods can present challenges that are addressed by the aqueous nature of capillary zone electrophoresis (CZE)-based separations. Therefore, if you have a choice of using LC-MS or CESI-MS when it comes to complex or polar molecules, the latter is ideal for your testing purposes since you can see and get high-resolution separation of PTMs missed by LC-MS methods. Such is the case for large molecules like intact monoclonal antibodies, and for small molecules, CESI can separate geometric isomers and isobaric compounds prior to mass spec analysis.
Want to see how CESI-MS plays out in the lab? View the Poster and get to know how molecules can be separated to differentiate their subtle structural changes. Collectively you’ll discover how the examples illustrate the simplicity of buffer systems that allow for the analysis of a wide range of highly relevant polar molecules by CESI-MS like phosphonate/phosphate for herbicide analysis to metabolites, oligosaccharides, and intact proteins.
What is more, you will also discover how you can improve the sensitivity, speed, and comprehensiveness of polar biomolecule analysis such as the separation of polar herbicide glyphosate and fungicide fosetyl aluminum. It’s not just polar ions the researchers put to the test either. Read about the results of anionic metabolites, glycans, and intact proteins too.
Find out how you can save time and resources using CESI-MS technology and read more about CE technology >
Finding the right information shouldn’t slow you down. Whether you’re troubleshooting your mass spec, learning something new, or optimizing performance, access to the right resources at the right moment makes all the difference.
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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