Forced degradation analysis of recombinant adeno-associated virus serotype 8 based on analytical anion exchange chromatography coupled to orthogonal characterization

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NIST26 introduces integrated chromatogram processing, deconvolution, and library searching for EI GC-MS and LC-MS/MS data, but the workflow begins with one essential requirement: chromatographic data must be available in the mzML format. In his presentation <a target="_blank" rel="noopener noreferrer" href="https://labrulez.com/pdf/Conversion_to_mz_ML_63e134bb34.pdf">Conversion to mzML Format for NIST26 Chromatogram Processing</a>, James Little of Mass Spec Interpretation Services explains how vendor data can be converted to mzML using either instrument-vendor software or the freely available MSConvert tool from ProteoWizard, and highlights several settings that are critical for successful processing in NIST26.<h4>NIST26 Chromatogram processing requires mzML</h4>For the chromatogram-processing workflow described in the presentation, mzML is the accepted input format. Proprietary LC-MS or GC-MS data therefore need to be converted before they can be loaded into the NIST26 chromatogram-processing environment.There are two principal ways to obtain an mzML file:<ul><li>conversion using software supplied by the instrument manufacturer,</li><li>conversion using MSConvert, which is included in the free ProteoWizard software package.</li></ul>Using an open format such as mzML also provides a practical way to move mass spectrometry data between different software environments without relying exclusively on proprietary vendor formats.<h4>Using ProteoWizard and MSConvert</h4>MSConvert is installed as part of the complete ProteoWizard package rather than as a separate application. After installing ProteoWizard, users can access either the graphical MSConvert interface or the command-line version.The presentation recommends the stable release for routine work, while noting that newer or nightly builds may sometimes provide improved support for recently introduced vendor formats or additional control over data-processing filters. This can be particularly relevant when handling vendor files in which centroiding behavior or peak picking requires closer attention.For most users, the graphical interface provides a straightforward route from proprietary raw data to an mzML file suitable for further NIST26 processing.<h4>Start by selecting mzML as the output format</h4>The first step in MSConvert is to select the source file or dataset using the Browse function.The software supports a broad range of mass spectrometry data formats, and the file-selection window can either be restricted to a particular vendor format or set to display any supported spectral format.After selecting the source data, the output format must be set to mzML. The screenshots in the presentation show this setting directly in the MSConvert options panel before the conversion is started.The user can also select the destination folder and other file-output options before adding the selected dataset to the conversion queue.<h4>Add the data and start the conversion</h4>Once the source file and output settings have been selected, the file is added to the conversion list using the Add command.The conversion itself is then launched with Start.For simple datasets that are already stored in the correct spectral representation, this may be all that is required. However, James Little emphasizes an additional requirement that is particularly important for NIST26: the data must be centroided.<h4>Centroided data are required</h4>According to the presentation, profile-mode data are not acceptable for this NIST26 chromatogram-processing workflow. The spectra must instead be in centroided, or peak-picked, form.Profile spectra contain the full measured shape of each mass spectral signal across the m/z axis. Centroided data reduce these profiles to discrete peaks represented by their characteristic m/z and intensity values.If the original vendor data are still in profile mode, centroiding can be performed during conversion by adding the Peak Picking filter in MSConvert.This means that the conversion step can perform two operations at once:vendor raw data → centroiding / peak picking → mzMLThe resulting mzML file can then be used as input for NIST26 chromatogram processing.<h4>Peak Picking must be the first filter</h4>One of the most important practical details in the presentation is the position of the Peak Picking operation in the MSConvert filter sequence.If centroiding is required, Peak Picking must be the first algorithm or filter in the list. Placing another processing function before it can interfere with the intended conversion workflow.MSConvert allows the user to select the Peak Picking filter, define the relevant MS levels, and add it to the processing sequence before starting conversion. The example shown in the presentation applies vendor-based peak picking to selected MS levels.If there is uncertainty about whether the source data are already centroided, the Peak Picking filter can still be included. According to the presentation, applying it to data that are already centroided does not alter the file unnecessarily.This provides a useful safeguard when the acquisition mode of an older dataset is uncertain.<h4>Selecting the appropriate MS levels</h4>Peak picking does not necessarily have to be applied identically to every spectrum in a dataset. MSConvert allows users to specify the MS levels to which the filter should be applied.This is especially relevant for LC-MS/MS data, where a file can contain both precursor-level MS spectra and tandem MS spectra.The conversion settings should therefore reflect the way the original data were acquired and how those data will subsequently be processed in NIST26.<h4>Be aware of changes in file size</h4>The presentation also notes that converted files may become substantially different in size from the original vendor files.This is worth considering when working with large LC-MS/MS datasets or processing batches of chromatograms. The size of the resulting mzML file can depend on the original acquisition mode, centroiding, compression settings, the number of MS levels, and other output parameters.Storage requirements should therefore be considered before converting large collections of raw data.<h4>A simple workflow for preparing data for NIST26</h4>The overall procedure presented by James Little can be summarized as a short sequence:<ol><li>Obtain the original vendor LC-MS or GC-MS data.</li><li>Open the data in MSConvert or use the vendor's own conversion software.</li><li>Select mzML as the output format.</li><li>Check whether the spectra are centroided.</li><li>If necessary, add Peak Picking as the first processing filter.</li><li>Select the appropriate MS levels.</li><li>Add the file to the conversion list.</li><li>Start the conversion.</li><li>Use the resulting mzML file for NIST26 chromatogram processing.</li></ol>The procedure itself is straightforward, but the centroiding requirement is important. A file can have the correct <code>.mzML</code> extension and still be unsuitable for the workflow if its spectra remain in profile mode.<h4>From vendor data to integrated NIST26 processing</h4>File conversion may appear to be a relatively minor step compared with deconvolution, spectral interpretation, and library searching, but it establishes the foundation for the entire downstream workflow.By using mzML as the common input format, NIST26 can process data originating from different instrument platforms through a more consistent workflow. ProteoWizard and MSConvert provide a practical bridge between proprietary mass spectrometry files and this standardized format.The key message from the presentation is therefore simple: convert to mzML, ensure the spectra are centroided, and place Peak Picking first whenever centroiding is required. Once those conditions are met, the data are ready for the integrated chromatogram-processing and library-searching capabilities available in NIST26.

Conversion to mzML Format for NIST26 Chromatogram Processing

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<div>Downstream process development usually requires numerous iterations until optimization is achieved. Offline analytics are often a bottleneck for timely development. The antidote to lagging offline analytics is inline or online process analytical technology (PAT).Real-time multi-angle light scattering (RT-MALS) is a PAT that monitors fundamental biophysical product attributes like molar mass, size and titer, every second during downstream operations such as FPLC or UF/DF. For AAVs, RT-MALS also monitors the full capsid ratio. This webinar describes the technology of RT-MALS and its implementation with a variety of bioprocess unit operations and modalities, from proteins to viral vectors, LNPs and pneumococcal vaccines.<h4>In this webinar, you will learn:</h4><ul><li>What are the CQAs and other attributes determined by RT-MALS for different modalities</li><li>How RT-MALS accelerates bioprocess DSP development</li><li>How process control with RT-MALS can enhance yield and lot consistency</li></ul><h4>Who Should Attend:</h4><ul><li>PAT experts for biologics, vaccines and gene therapies</li><li>Biopharm process development scientists</li><li>Downstream bioprocess engineers</li><li>Biopharm MSAT leaders</li></ul><blockquote>Presenter: Dan Some, Ph.D. (Sr. Principal Scientist, Waters | Wyatt Technology)Dr. Daniel Some is a Sr. Principal Scientist at Waters | Wyatt Technology, where he contributes to product and application development as well as scientific and technical marketing. Currently he leads Wyatt’s program for commercializing real-time MALS PAT products.</blockquote></div>

Waters Corporation: Accelerating Downstream Process Development for Biologics, Gene Vectors and Vaccines

Accelerating Downstream Process Development for Biologics, Gene Vectors and Vaccines

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Accelerating Downstream Process Development for Biologics

Gel Permeation Chromatography/Size Exclusion Chromatography (GPC/SEC) is a non-interactive chromatographic technique that separates polymer samples based on their size in solution, rather than their chemistry. &nbsp; Developing a GPC/SEC method involves several key steps to ensure accurate and reproducible results.In this e-Seminar, we will explore the key parameters of GPC/SEC method development:<ul><li>Identify the Polymer Sample: Understanding the type of sample.</li><li>Solvent Choice: Deciding between organic and aqueous solvents.</li><li>Column Selection:&nbsp; Considerations for packing type, particle size, and pore size. &nbsp;Learn how to maximize column performance through selection.</li><li>Calibration: Choosing appropriate standards and understanding their significance.</li><li>Optimize the System: &nbsp;Considerations for flow rate, temperature, and injection volume</li><li>Detection: Utilizing refractive index (RI) and advanced detection methods.</li></ul>We will delve into each of these areas in detail, offering technical insights, application examples, and share valuable tips to help you develop a GPC/SEC method that is both robust and reproducible. &nbsp;<blockquote>Presenter: Jean Lane (Application Engineer, Agilent Technologies, Inc.)Jean has been employed with Agilent since Nov 2010 in technical and applications support and has a primary role supporting Agilent’s full line of LC columns and consumables, which also includes support for Agilent’s GPC/SEC product line. She came to Agilent thru acquisition of Varian and the former Polymer Laboratories. Jean has 20 plus years of technical and applications support for both HPLC and GPC consumable products. In addition to practical chromatography experience in the laboratory, she has had many, many years in her technical role involving customer support providing assistance and troubleshooting for customer chromatography issues.</blockquote>

Agilent Technologies: A Systematic Approach to GPC/SEC Method Development

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In Part 1 of this workshop, we discuss the capabilities of the Praying Mantis accessory from Specac/Harrick in combination with the Cary 5000 UV-VIS/NIR System. &nbsp;It is a critical accessory for assessing the diffuse reflectance of small volumes of powders. &nbsp;Additional sampling capabilities through specialized reaction chambers allow for the control of sample pressure, temperature, and environment for catalyst studies.<blockquote>Presenter: Scott Melis, PhD (Application Scientist, Agilent Technologies, Inc.)Scott Melis has a PHD in Physics from Georgetown University in Washington, DC and started with Agilent in 2021 as a Molecular Spectroscopy Application Scientist. In his graduate work, Scott studied nanoparticle formation and growth for a variety of applications. Projects that he worked with include developing processes to deposit and characterize semiconducting molecular nano-cocrystals for use in optoelectronic devices as well as controlling polymer nanoparticle size through rapid mixing conditions.</blockquote>

Agilent Technologies: Optimize your FTIR, UV-Vis-NIR, Raman, Imaging, and Fluorescence Workflows

Part 1: Characterizing catalysts using an UV-VIS/NIR system with the Praying Mantis diffuse reflectance accessory.

Part 1 discusses the capabilities of the Praying Mantis accessory (Specac/Harrick) in combination with Cary 5000 UV-VIS/NIR System.

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<h4>Preparing for next generation LC-MS based proteomics</h4>Presenter: Nicolai Bache (VP Proteomics research at Evosep)

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Get a sneak peek of what’s coming at ASMS 2025.

<h4>Expand Your Expertise with Our Chromatography Webinar Series 2025</h4>A comprehensive journey through fundamental principles, HPLC optimization, and cutting-edge mixed-mode techniques. Led by industry experts, these free sessions provide practical insights and real-world applications to enhance your analytical skills.<h5>Session 3: Introduction to Mixed-Mode Chromatography in HPLC</h5><ul><li>What is mixed-mode chromatography?</li><li>Advantages over single-mode techniques (RP, IEX, HILIC, etc.)</li><li>Mechanisms of multiple interactions: hydrophobic, ionic, hydrogen bonding, and π-π interactions</li><li>How mixed-mode selectivity can improve complex separations</li></ul><blockquote>Presenter: Jovana Mitrovic (R&amp;D Chemist at SIELC Technologies)Jovana holds a Master of Science in Molecular Engineering from the Pritzker School of Molecular Engineering at the University of Chicago and a Bachelor of Science in Biomedical Sciences from St. John’s University. Her background in molecular and biomedical sciences supports a multidisciplinary approach to addressing complex analytical challenges.</blockquote>

SIELC Technologies: HPLC Educational Webinar Series

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Join us for a webinar and learn about what is mixed-mode chromatography, advantages over single-mode techniques (RP, IEX, HILIC) and other useful things.

<h4>An introduction to accurate mass and Quadrupole-Time-of-Flight technology with SCIEX</h4>Until recently, nominal mass triple quadrupole technology was considered the gold standard for quantitative assays. In contrast, high-resolution mass spectrometry was primarily used for qualitative, semi-quantitative, and non-targeted workflows, such as metabolite identification, screening, and X-Omics. However, advancements in technology, including faster scan speeds and improved ion optics, have expanded the capabilities of high-resolution spectrometry. Now, it can be effectively applied to both qualitative and quantitative workflows.&nbsp;The ZenoTOF 7600 system, with its high scan rates and enhanced duty cycle at the MS2 level, bridges the gap between these approaches, offering a greater return on investment by reducing data acquisition times and maintaining sensitivity without compromising data quality. This webinar series will focus on how to transition from the Triple Quad to a high-resolution environment for different applications.This webinar series will contain 3 parts. Each part will be 20 minutes with a 10 minute Q&amp;A.<h4>Part 3: Alternative Fragmentation Modes – EAD, CID and more</h4><ul><li>What are EAD &amp; other alternative fragmentation modes and how to do they work?</li><li>What do EAD spectra look like for different molecule types?</li><li>Benefits of EAD and when to use it with or instead of CID</li></ul><blockquote>Presenter: Tom Ruane (Accurate Mass Workflows Specialist, SCIEX)Tom has been working for SCIEX since 2018, having worked at Phenomenex since leaving the University of Manchester in 2015. He currently helps SCIEXs customers across Nordic countries, UK and Ireland working in life science research and drug development. In particular he has an interest in accurate mass technologies.</blockquote>

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Get ready for a game-changing way to efficiently extract even more information from your molecules. Become a mass detection pro and drive higher productivity with less effort. &nbsp;Join us for this must-see virtual launch event and be the first to learn about the newest innovative solution from Agilent.Discover how you can:<ul><li>Break through the limits of traditional methods</li><li>Deliver results confidently</li><li>Radically simplify user training and daily operation</li></ul>Whether you are a part of a research, development, or QA/QC analysis group, Agilent is ready to take your lab to the next level.Don’t miss your chance to ride the wave of innovation.

Agilent Technologies: Ride the Wave of Innovation

Ride the Wave of Innovation

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<div><div container-id="cq-colctrl-lt0" comp-id="A1"><div><h4>Lessons learned in developing an ion pair free, compliant workflow for ASO RNA impurity analysis</h4>Join this live webinar with Dr. Ken Cook and discover innovative techniques for oligonucleotide impurity quantitation, automation, and data handling to enhance your laboratory's efficiency and compliance.<h5>Topics:</h5><ul><li>Ion pair free separation mechanisms<ul><li>Reasons for choosing the best path for quantitative methods.</li></ul></li><li>Removing problems of adducts and in-source induced impurities<ul><li>Preventing accurate quantitation of real impurities.</li></ul></li><li>Choice of the best charge states to use for quantitation</li><li>Deconvolution and XIC Component Tables in Thermo Scientific Chromeleon Software<ul><li>Compliant data handling and complementary techniques.</li></ul></li><li>Automation of Annotation, Integration, and Compliant Reporting<ul><li>UV and MS data handling to remove bottlenecks in developing and running new quantitative methods for oligonucleotide impurity analysis</li></ul></li></ul>After the presentation, we planned time to answer your questions.<h5>Who should attend?</h5><ul><li>Laboratory managers or analysts<ul><li>developing methods for new synthetic therapeutic oligonucleotides</li><li>implementing methods for existing modalities</li></ul></li><li>Laboratories<ul><li>involved in clinical trial analysis</li><li>aiming to avoid ion pair contamination in their LC-MS systems</li></ul></li><li>Oligonucleotide analysts in R&amp;D or QC departments</li><li>Analysts looking for green oligonucleotide chemistry</li></ul>You cannot attend the live broadcast on May 22? Please register to automatically receive an email with the link to the recording.<blockquote>Presenter: Dr. Ken Cook (Applications Scientists Manager, Thermo Fisher Scientific)Moved to industry from a University lectureship in Biochemistry, University of Newcastle upon Tyne, following 7 years’ experience in academia. Now with over 35 years’ experience with Thermo Fisher Scientific using a wide range of analytical equipment and software. Publications range from metabolomics, proteomics to large biopharmaceutical drug analysis. Current job title is the European manager for the BioPharma expert support group. This involves the support of Bio-Pharmaceutical applications to characterise protein and oligonucleotide based bio-therapeutics. This involves ongoing collaborations with the Biopharmaceutical industry and academia. Characterization methods of Bio-therapeutics include, monoclonal antibodies, peptides, AAV analytical development, small and large RNA analysis including sequence mapping of mRNA by LC/HRMS.</blockquote></div></div></div>

Thermo Fisher Scientific: ASO RNA impurity analysis

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Discover innovative techniques for oligonucleotide impurity quantitation, automation, and data handling to enhance your laboratory's efficiency and compliance.

<div><div container-id="cq-colctrl-lt0" comp-id="A1"><div>Halogens like fluorine, chlorine, bromine, and iodine, along with sulfur, are corrosive and hazardous elements that pose significant environmental and industrial challenges. Accurate monitoring of these elements is crucial to protecting human health and ensuring efficient manufacturing processes. Traditional methods often struggle to detect these analytes in complex samples such as petrochemicals and personal care products, complicating quality control and assurance efforts.Join us for an exclusive product launch webinar where three experts from Thermo Fisher Scientific will introduce an innovative Combustion-Ion Chromatography (C-IC) system. This cutting-edge technology is designed to enhance your halogen analysis, streamline your workflow, improve data accuracy, and deliver exceptional precision and efficiency.Gain valuable insights from Dr. Carl Fisher, Pavan Sandhu, and Dr. Neil Rumachik as they discuss the advanced features, operational benefits, and application performance of this new C-IC system.<h5>Key learning objectives</h5><ul><li>Gain a comprehensive understanding of the features and benefits of the new C-IC system</li><li>Explore the enhanced capabilities and diverse applications of the system across various industries</li><li>Discover how this system can optimize your workflow and significantly improve data accuracy</li></ul><h5>Who should attend?</h5><ul><li>Laboratory managers, directors, quality control and assurance professionals</li><li>Environmental scientists, analytical chemists, technicians and researchers</li><li>Professionals in the consumer product manufacturing, power generation, food, and chemical industries</li></ul><h4>In a different time zone?</h4>This webinar is one of three sessions that are taking place over different time zones. This webinar has been specifically scheduled for the European time zone. Please use the links provided to access the session that best suits your region:<ul><li><a target="_blank" rel="noopener noreferrer" href="https://www.selectscience.net/webinar/spark-new-insights-boost-your-lab-s-power-with-combustion-ion-chromatography">USA</a></li><li><a target="_blank" rel="noopener noreferrer" href="https://www.selectscience.net/webinar/spark-new-insights-boost-your-lab-s-power-with-combustion-ion-chromatography-asia">Asia</a></li></ul><h5>Certificate of attendance</h5><ul><li>All webinar participants can request a certificate of attendance, including a learning outcomes summary, for continuing education purposes.</li><li>If you view the on-demand webinar, you can request a certificate of attendance by emailing editor@selectscience.net.</li></ul><blockquote>Presenter: Dr. Carl Fisher (Product Marketing Manager, Thermo Fisher Scientific)Dr. Carl Fisher is a Product Marketing Manager at Thermo Fisher Scientific. He earned his PhD in Biochemistry from the University of Alberta in Edmonton, AB, which was followed by a post-doctoral fellowship at Brigham and Women’s Hospital, Harvard Medical School in Boston, MA. After joining Thermo Fisher Scientific, he spent several years in the Applications Development team as an analytical chemist, producing technical and application notes. In his current role, he drives the delivery of Ion Chromatography and sample preparation solutions for the environmental and industrial markets.Presenter:&nbsp;Pavan Sandhu (Product Management Specialist, Thermo Fisher Scientific)Pavan Sandhu completed her MS in Biomedical Engineering from University of Texas at Dallas and has several years of experience in clinical diagnostics and analytical instrumentation. At present, she is the Product Management Specialist in the Ion Chromatography &amp; Sample preparation (IC/SP) business unit at Thermo Fisher Scientific, managing Combustion IC &amp; Discrete Analyzer products. Prior to joining IC/SP, she was part of an R&amp;D team at Thermo Fisher’s Clinical Diagnostic Division (CDD) leading product development and design transfer of multiple diagnostic products.Presenter:&nbsp;Dr. Neil Rumachik (Applications Development Manager, Thermo Fisher Scientific)Dr. Neil Rumachik completed his graduate studies in Chemistry at the University of California, Berkeley (MS), and Stanford University (PhD) under the mentorship of Professor Carolyn Bertozzi before joining Thermo Fisher Scientific. He currently works as the Manager of Applications Development in the Ion Chromatography and Sample Preparation business unit at Thermo Fisher Scientific. Before moving to the apps team, he worked as a staff scientist in R&amp;D at Thermo Fisher, where he developed IC-MS products and workflows for carbohydrate analysis. With over a decade of experience at the intersection of chemistry and biology, he is focused on developing cross-disciplinary methods and utilizing multi-omics platforms to advance biological studies.Moderator: Cameron Smith-Craig (Pharma and Applied Sciences Editor, SelectScience)</blockquote></div></div></div>

Thermo Fisher Scientific: Spark new insights - Boost your lab's power with combustion-ion chromatography

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Experts from Thermo Fisher Scientific will introduce an innovative Combustion-Ion Chromatography (C-IC) system. 

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This month, our seminar series features cutting-edge proteomics research from leading scientists in Canada and the U.S. We are pleased to welcome Dr. Jennifer Geddes-McAlister (University of Guelph), who will discuss a proteomics-driven approach to fungal disease, from prevention to treatment, and Dr. Ben Garcia (Washington University in St. Louis), who will present on using quantitative proteomics to investigate epigenetic mechanisms in human health. Don’t miss this exciting seminar — register for free today!<blockquote>Presenter: Dr. Jennifer Geddes-McAlister (University of Guelph)<ul><li>Talk: From prevention to treatment: a proteomics-driven approach to fungal disease</li></ul>Presenter: Dr. Ben Garcia (Washington University)<ul><li>Talk: Quantitative proteomics for understanding epigenetic mechanisms in human disease</li></ul></blockquote>

US HUPO: US HUPO-CNPN Proteomics Spotlight

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