With 3 decades of experience in world-class healthcare expertise, Intas is one of the leading global pharmaceutical formulation development, manufacturing and marketing companies headquartered in India. Intas strives to learn and lead with a steadfast commitment to provide efficient and affordable health to all.
NMR spectroscopy has a unique capability to probe the primary and higher order molecular structure and also the structural dynamics of biomolecules at an atomic resolution. ICH Topic Q6B includes the use of NMR as a tool for understanding the higher order structure of biopharmaceuticals.
Due to improvements in instrumentation and methodologies, increased structural information can be obtained from smaller amounts of sample than previously required and NMR is now used more widely and frequently in the characterization of biopharmaceuticals and development of biosimilars.
Intas required the characterization of several samples of a pegylated GCSF molecule using 1-Dimensional (1D) 1H-NMR and 2-Dimensional (2D) 1H/13C-NMR at 800MHz. In addition, Intas requested that BioPharmaSpec purify the PEGylated N-terminal peptide from a representative sample in sufficient quantities to allow 1D 1H-NMR and 2D 1H/13C-NMR analysis to provide an assessment of the linkage between PEG/Linker and the N-terminal Methionine residue.
Technical considerations- NMR Spectroscopy
BioPharmaSpec recommends NMR analysis of biopharmaceuticals in order to provide a high-end assessment of higher order structure, as well as probe certain key aspects of the primary sequence. The US FDA has also stated in numerous presentations that NMR is the technique used in their laboratories to provide a detailed assessment of HOS.
BioPharmaSpec provides 1D 1H-NMR data for an initial assessment (and for analysis of products at too low a concentration for 2D-NMR) followed by 2D-NMR (normally 1H – 13C 2D NMR analysis). BioPharmaSpec recommends 1H – 13C 2D-NMR over 1H – 15N 2D-NMR because of the higher natural abundance of the 13C isotope which provides a more significant response, particularly for products larger than 50kDa.
During product development, BioPharmaSpec recommends a combination of CD near and far-UV, FT-IR, NMR and Intrinsic or Extrinsic Fluorescence to assess the higher order structure of a biopharmaceutical. The combination of these tests provides the breadth and orthogonality required by the EMA and US FDA.
“At Intas, we are committed to characterizing our products to the highest standard and to producing quality products. BioPharmaSpec were able to offer a high-end technique and excellent technical support and guidance around the data interpretation. This adds significant value to our development product and helps us achieve our overall aim of providing efficient and affordable healthcare” – Senior General Manager, Intas Pharmaceuticals Ltd.