Overcome Oligonucleotide Analysis Challenges with 2D-LC/MS

With the enormous potential of oligonucleotide therapeutics, there is more interest than ever in choosing the best tools for oligonucleotide synthesis, analysis, and purification. Oligonucleotides synthesized using phosphoramidite chemistry are typically analyzed and purified using ion-pair reversed-phase liquid chromatography (IP-RPLC) and anion-exchange chromatography. Since highly salty mobile phases are not compatible with mass spectrometry, and samples need to be desalted to participate in ion-pairing prior to IP-RPLC analysis, investigators can either manually prepare samples or consider using 2D-LC/MS to obtain mass measurement from salty first dimension separations. 2D-LC increases the workflow speed and avoids manual sample preparation. Given the complex impurity profiles, varying sequence modifications and lengths, thoughtful optimization of HPLC methodologies for characterization and purification is essential to obtain pure oligonucleotides for research, diagnostics, and therapeutics.

This webinar will discuss the common challenges associated with oligonucleotide analysis and purification and how to overcome them through 2D-LC/MS and method optimization.

Key learning objectives

• Understand how to couple multiple LC methods for oligonucleotide detection into one seamless application
• Simplify transitions from 1D salty buffers to a 2nd dimension LC-MS separation
• Technical challenges and impurities associated with oligonucleotide synthesis
• Oligonucleotide analysis with varying oligonucleotide structures (size, modifications)
• Method optimization considerations for ion pair reverse phase purification and for MS analysis

Who should attend?

• Analytical lab managers
• Lab directors
• Anaylysts
• Oligonucleotide synthesists
• Academic researchers

Presenter: Patrick Cronan (LC applications scientist, Agilent Technologies)

Patrick Cronan is an LC applications scientist with Agilent Technologies. He is an accomplished analytical chemist with over 20 years of experience in the pharmaceutical and biopharma industries and has facilitated method transfer from HPLC to UHPLC while using LC-MS to characterize large and small molecule drug-like compounds. Patrick has also worked extensively with Agilent’s 2D chromatography system, developing methods to increase throughput of biologic separations by coupling multiple techniques like SEC x IEX.

Presenter: Matthew L. Turner, Ph.D. (Biocolumns product manager, Agilent Technologies)

Matthew Turner is a product manager of Biocolumns at Agilent Technologies, Inc., supporting the cell and gene therapy market. Previously Matthew worked as a field applications scientist for four years supporting cell and gene therapy customers in their genomic and proteomic analysis and lab scale purification workflows. Matthew received his Ph.D in biochemistry and structural biology from the University of California, Davis in 2016, with a focus on structural neuronal proteins and their effect on synaptic plasticity, before pursuing his career supporting biopharma customers.

Presenter: Moderator: Sarah Thomas (Editorial Team, SelectScience)

Sarah studied biology at the University of Bath, UK, and has worked in the Pharmaceutical Sciences Department of St. Jude Children’s Research Hospital, Memphis. As a member of the Editorial team, Sarah plays an integral role in shaping the content on SelectScience.