A Holistic Inert LC/MS Workflow for Enhanced Characterization of Peptides and Oligonucleotides
Date: Wednesday, 12 November
Time: 11:50-12:20
Dive into the latest breakthroughs in oligonucleotides, peptides, mRNA, drug delivery, and strategic partnerships—from early innovation to commercial success.
Connect with our team to explore innovative technologies and streamlined workflow solutions designed to support and accelerate your biopharma journey.
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What to expect:
The presentation introduces a holistic LC solution combining the Infinity III BioLC system and inert column technology to overcome metal sensitivity challenges in biotherapeutics, enhancing sensitivity and peak shape with next-gen Pro iQ instrumentation.
Join us at booth #317 and experience innovation in action!
Connect with the Agilent team and explore transformative technologies that are shaping the future of science. And don’t miss the poster sessions, where you’ll get an exclusive look at the latest advancements in the Agilent Oligo Analysis Accelerator software, and the new Agilent Altura HPLC Columns.
This poster presents an automated LC/MS workflow using the Agilent 6170A Pro iQ Plus system and Oligo Analysis Accelerator software to streamline ASO impurity profiling and purity assessment, reducing manual effort and enhancing QC reliability.
Metal-induced analyte loss is a key challenge in LC/MS, especially for peptides and oligonucleotides. This study highlights Altura, an inert LC column with coated hardware, delivering up to 25x stronger signals and improved quantitation.
Discover the Agilent Pro iQ - the next generation in LC-Mass Detection.
With the Pro iQ Series, Agilent empowers your lab to achieve more—faster, smarter, and with greater confidence.
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Presented by: Dr. Holger Stalz, LCMS Product Specialist and Dr. Anne Blackwell, Biocolumns & Consumables Product Manager
Emerging biotherapeutic modalities increase analytical complexity, posing additional challenges to robust and sensitive methods. These molecules are often metal-sensitive, resulting in sub-optimal chromatography and limited analytical sensitivity. We will present a holistic approach, introducing the Infinity III BioLC system paired with novel inert column technology that mitigates metal interactions to improve peak shape and increase sensitivity, driven by the advanced intelligence capabilities of next-generation Pro iQ single quad instrumentation.
Anti-sense oligonucleotides (ASOs) are a rapidly growing class of therapeutic agents, requiring precise and reliable analytical methods for quality control (QC). Liquid chromatography-mass spectrometry (LC/MS), particularly single quadrupole detection, has emerged as a robust technique for assessing ASO purity. However, traditional chromatography data systems often demand extensive manual intervention, especially for tasks such as extracted ion chromatogram (EIC) integration and impurity profiling.
This study demonstrates an automated LC/MS workflow using the Agilent 6170A Pro iQ Plus LC/MS system in combination with the Oligo Analysis Accelerator (OAA) software for OpenLab CDS. The system enables high-sensitivity detection across an extended mass range (2–3000 m/z), allowing for accurate identification and quantitation of co-eluting impurities. The OAA software streamlines complex analytical steps, including ion classification, EIC integration, and UV peak alignment, through a guided interface and API-driven automation. Using a 20mer ASO sample, the workflow achieved MS purity of 86.9% and UV purity of 98.0%, highlighting the importance of mass detection in identifying closely related impurities.
This integrated approach significantly reduces manual processing, enhances data integrity, and facilitates method transfer for routine QC in compliant environments. The results underscore the value of combining purpose-built software with advanced instrumentation to modernize oligonucleotide analysis.
Metal-induced analyte loss remains a critical barrier in LC and LC/MS workflows for biologically relevant molecules, particularly peptides and oligonucleotides. These analytes exhibit strong affinity for metal surfaces, leading to signal suppression, poor peak shapes, and compromised quantitation.
This study presents a comprehensive evaluation of Altura, a newly developed inert LC column platform, comprised of coated stainless steel hardware, with a focus on phosphopeptides. Inert column hardware was confirmed to be robust under extreme operating conditions, as well as reproducible across batches. In combination with an inert LC, columns can improve peak area of metal-sensitive analytes by up to 25x versus a traditional LC system with a stainless steel column. Even on a traditional LC system, peak area increased up to 14x for metal sensitive columns simply by switching from a SS column to an inert column.