Using qNMR as Platform Methods of Oligonucleotides and Peptides

The content of peptides and oligonucleotides in a sample (solid or liquid) is important in many aspects. For peptides, traditional methods like Kjeldahl nitrogen determination, elemental analysis (EA), and amino acid analysis (AAA) require breaking down the protein under harsh conditions to determine its content.

For oligonucleotides, one of the most widely used quantification techniques is ultraviolet (UV) spectroscopy. This method involves time-consuming sample preparation and, due to uncertainties in the extinction coefficient, its accuracy is negatively affected.

NMR spectroscopy is inherently quantitative, and quantitative NMR (qNMR) has been successfully applied in small molecule analysis for decades. However, for larger molecules like peptides and oligonucleotides, the quantitative aspect of 1H-NMR is hindered by severe line broadening caused by higher-order structural elements like dimers or oligomers under native conditions (e.g., peptides).

This webinar will explore the application of analyzing peptides in non-native conditions (a deuterated water:acetonitrile mixture) and recording 1H-NMR data at elevated temperatures.

For oligonucleotides, the use of 1H-NMR is not a viable option, and data will be presented with 31P as the selected nuclei for qNMR. Within the two modalities (peptides and oligonucleotides), the validation of the methods was performed using the recommendations from both USP and ICHQ2(R2) with respect to validation parameters (e.g. linearity) and platform applicability.

The content results obtained for both peptides and oligonucleotides will be compared to the content results obtained by the traditional methods.

Key Topics

  • Quantitative determination of peptides and oligonucleotides using qNMR
  • Method validation according to USP and ICHQ2(R2) guidelines
  • Assessment of platform applicability for qNMR Methods

About the Speakers

Joan Malmstrøm, a Principal Scientist at Novo Nordisk A/S, earned her Ph.D. in organic chemistry from the University of Copenhagen. She brings over 25 years of expertise in structural elucidation of organic molecules, utilizing nuclear magnetic resonance (NMR) spectroscopy and mass spectrometry. At Novo Nordisk A/S, her work has encompassed a diverse range of projects, including small molecules (APIs, raw materials, and excipients), as well as peptides, proteins, oligonucleotides, and large molecular weight polymers. In recent years, she has concentrated on developing NMR analytical methods for various molecules involved in drug development. These methods are implemented throughout all stages of the development pipeline and are GMP-validated when required.

Victor has extensively studied and applied NMR techniques during his undergraduate years at the University at Buffalo under Dr. Thomas Szyperski, and throughout his Ph.D. program at Yale University with Dr. Patrick Loria. He subsequently joined Pfizer as an NMR specialist focused on the structural characterization of biotherapeutics before transitioning to project management in analytical laboratory automation. His passion lies in enhancing quality of life and operational efficiency by driving high-quality scientific innovation, particularly through state-of-the-art NMR characterization of novel therapeutic modalities. At Bruker, he collaborates with scientists to understand industry challenges and needs, aiming to broaden the applications and technology of NMR.

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