Using NMR Spectroscopy to Obtain the Higher Order Structure of Biopharmaceutical Products - Simple methods can characterize polysaccharide vaccines and recombinant cytokines at high resolution. - BioP


Using NMR Spectroscopy to Obtain the Higher Order Structure of Biopharmaceutical Products
Simple methods can characterize polysaccharide vaccines and recombinant cytokines at high resolution.

BioPharm International Supplements


Although NMR instruments have a relatively high initial capital outlay, all major manufacturers of polysaccharide and conjugate vaccines now use NMR spectroscopy to assess the identity and purity of the saccharide component, and NMR is a pharmacopoeial identity test for heparin. It also is proposed as an identity test for peptide hormones. In addition, companies specialized in providing analytical services to small biotech companies may see opportunities by investing in NMR hardware or purchasing spectrometer time at NMR laboratories on university campuses that already offer NMR services to local industries. The increasing availability of these instruments in biopharmaceutical manufacturing environments will allow them to be further exploited for tests that would otherwise not justify the resources. The growth in the use of NMR spectroscopy in biopharmaceutical quality control is influencing the acceptance and even expectation of these data by regulatory authorities. The analytical challenges of biosimilars have increased the need for analytical methods able to distinguish small structural differences between molecules. NMR spectroscopy meets this criterion, and provides one of a limited number of methods able to provide detailed information on higher order structure.


The authors thank Cangene for generously providing samples of Leucotropin and Fabian Jameison, PhD, of the US Pharmacopeia for providing goserelin acetate and goserelin-related compound A.

YVES AUBIN leads the NMR laboratory at the Protein Structure and Analysis Laboratories at BGTD, Health Canada, Ottowa, Canada, 613.941.6155,
CHRISTOPHER JONES is the division head at the Laboratory for Molecular Structure at the National Institute for Biological Standards and Control, Herts, UK, and DARÓN I. FREEDBERG leads the NMR laboratory at the Center for Biologics Evaluation and Research, US Food and Drug Administration, Rockville, MD.


1. Abeygunawardana C, Williams TC, Sumner JS, Hennessey JP. Development and validation of an NMR-based identity assay for bacterial polysaccharides. Anal Biochem. 2000;279:226–40.

2. Cavanagh J, Fairbrother W, Palmer A, Rance M, Skelton N. Protein NMR Spectroscopy, 2nd Ed. Academic Press: San Diego, 2007.

3. Frasch CE, Concepcion N. Induction of group 17 specifi antibodies by pneumococcal type 17F and 17A polysaccharide vaccines. Biologicals. 2001;29(1):11–16.

4. C. Jones. Vaccines based on the cell-surface carbohydrates of pathogenic bacteria. An Bras Acad Cienc. 2005;77:293–324.

5. Mawas F, Niggemann J, Jones C, Corbel MJ, Kamerling JP, Vliegenthart JFP. A conjugate vaccine made with a synthetic single repeating unit of Type 14 pneumococcal polysaccharide coupled to CRM197 is immunogenic in a mouse model. Infect Immun. 2002;70:5107–114.

6. Kellenbach E, Sanders K, Overbeeke PLA. The use of proton NMR as an alternative for the amino acid analysis as identity test for peptides. In: Holzgrabe U, Wawer I, Diehls B, editors. NMR spectroscopy in Pharmaceutical analysis. Oxford, UK: Elsevier; 2008. p 429–36

7. Aubin Y, Gingras G, Sauvé S. Assessment of the three-dimensional structure of recombinant protein therapeutics by NMR fingerprinting: demonstration on rhGM-CSF. Anal Chem. 2008;80:2623–7.

8. Klaus W, Gsell B, Labhardt AM, Wipf B, Senn H. The three-dimensional high resolution structure of human interferon alpha-2a determined by heteronuclear NMR spectroscopy in solution. J Mol Biol. 1997;274:661–75.

9. Sauvé S, Gingras G, Aubin Y. NMR assignment of backbone and side chain resonannces for human granulocyte-macrophage colony-stimulating factor. Biomol NMR Assign. 2008;2:5–7.

10 . Panjwani N, Hodgson DJ, Sauvé S, Gingras G, Aubin Y. Assessment of the Effects of pH, Formulation, and Deformulation on the Conformation of Interferon alpha-2 by NMR. J Pharm Sci, Published Online: 23 Feb 2010, DOI 10.1002/jps.22105.

blog comments powered by Disqus



New Tax Rules May Deter Future Pharma M&A
October 1, 2014
NIH Seeks to Improve Vaccine Response with New Adjuvants
September 30, 2014
New Report Details Players and Pipelines in the Biosimilar Space
September 30, 2014
Baxter International Plans to Open R&D Center for Baxalta
September 30, 2014
FDA Releases First-Ever Purple Book for Biosimilar Characterization
September 26, 2014
Author Guidelines
Source: BioPharm International Supplements,
Click here