Mapping Monoclonal Antibody Structure by 2D NMR at Natural Abundance
Mapping the structure of monoclonal antibodies using 2D 13C and 1H-15N correlated NMR methods at natural abundance. Research focuses on obtaining structural maps and statistical comparability of mAbs, crucial for pharmaceutical industry's therapeutic development platforms. Interlaboratory study conducted to harmonize and validate NMR measurements for assessing higher order protein structure, with NIST-Fab system suitability sample serving as benchmark. Collaboration of 30 institutions from 10 countries across industry, academia, and government sectors.
Download Presentation
Please find below an Image/Link to download the presentation.
The content on the website is provided AS IS for your information and personal use only. It may not be sold, licensed, or shared on other websites without obtaining consent from the author.If you encounter any issues during the download, it is possible that the publisher has removed the file from their server.
You are allowed to download the files provided on this website for personal or commercial use, subject to the condition that they are used lawfully. All files are the property of their respective owners.
The content on the website is provided AS IS for your information and personal use only. It may not be sold, licensed, or shared on other websites without obtaining consent from the author.
E N D
Presentation Transcript
NIST Published data NIST Published data Please cite: 1) 13C data Arbogast, L.W.; Brinson, R.G.; Marino, J.P. Mapping Monoclonal Antibody Structure by 2D 13C NMR at Natural Abundance. Analytical Chemistry, 2015, 87, 3556 3561. 2) 15N data Arbogast, L.W.; Brinson, R.G.; Marino, J.P. 2D 1HN, 15N Correlated NMR Methods at Natural Abundance for Obtaining Structural Maps and Statistical Comparability of Monoclonal Antibodies. Pharmaceutical Research, 2016, 33,462-475
900 MHz mAb Amide Spectra 900 MHz mAb Amide Spectra 64 hrs 32 hrs Arbogast, L.W.; Brinson, R.G.; Marino, J.P. 2D 1HN, 15N Correlated NMR Methods at Natural Abundance for Obtaining Structural Maps and Statistical Comparability of Monoclonal Antibodies. Pharmaceutical Research, 2016, 33,462-475
mAb Domain Amide Spectra mAb Domain Amide Spectra 32 hrs 32 hrs 64 hrs 64 hrs Arbogast, L.W.; Brinson, R.G.; Marino, J.P. 2D 1HN, 15N Correlated NMR Methods at Natural Abundance for Obtaining Structural Maps and Statistical Comparability of Monoclonal Antibodies. Pharmaceutical Research, 2016, 33,462-475
NISTmAb Methyl Spectra NISTmAb Methyl Spectra 9 hrs 4.5hrs 9 hrs 9 hrs Arbogast, L.W.; Brinson, R.G.; Marino, J.P. Mapping Monoclonal Antibody Structure by 2D 13C NMR at Natural Abundance. Analytical Chemistry, 2015, 87, 3556 3561.
Domain Cleavage does not alter HOS Domain Cleavage does not alter HOS Red= Fab/Fc (2:1) Blue=Full length Overlay spectrum of the full length NISTmAb (blue) and the Fab/Fc (2:1) digest product (red). Arbogast, L.W.; Brinson, R.G.; Marino, J.P. Mapping Monoclonal Antibody Structure by 2D 13C NMR at Natural Abundance. Analytical Chemistry, 2015, 87, 3556 3561.
Interlaboratory Interlaboratory Study: Goals Study: Goals To harmonize and to validate NMR structural fingerprinting measurements for the assessment of higher order structure of large protein biologics and/or domains from these proteins. The validation of NMR methods for the characterization of the higher order structure of mAbs is specifically targeted due to the large interest of the pharmaceutical industry in using mAbs as platforms for therapeutic development. Samples U-15N, 20%-13C NIST-Fab system suitability sample. This NIST-Fab sample was expressed in P. pastoris at Health Canada in Ottawa, Ontario, Canada. This sample is serving as the benchmark for all measurements. Unlabeled NIST-Fab. This NIST-Fab sample was derived from the NISTmAb candidate reference material #8670. The unlabeled fragment was generated at NIST-IBBR in Rockville, MD, USA. A total of 30 collaborating institutions from 10 countries, with equal participation from industry, academia, and government. 6
