NIBSC have a wide variety of techniques for protein and glycoprotein characterisation which have been developed to support examination of biotherapeutic and vaccine products which are either licensed or for which licensed are being sought, and to provide support for the establishment of reference standards.

The available methods include:-

• HPLC and capillary electrophoresis separations of intact proteins
• One- and two dimensional- gel electrophoresis with a variety of staining approaches, including DIGE
• Peptide mapping by HPLC and capillary electrophoresis, and coupled nanoflow HPLC-ESI-MS-MS for identification and quantification.
• MALDI-TOF mass spectrometry and MALDI TOF-TOF mass spectrometry
• Fluorescence-detected HPLC and mass spectrometic analysis of glycoprotein glycans
• Gel permeation chromatography and GPC-MALLS for molecular weight determination and aggregation status
• Circular dichroism and fluorescence spectroscopy, for analysis of secondary structure, protein stability and protein interactions.
• Highfield NMR for peptide identification and quantification.

The goals of this work include:-

• Protein identification or verification of protein identity (purified proteins)
• Identification of protein components in complex mixtures
• Quantification of pure proteins and specific proteins in complex mixtures
• Protein purity (other proteins and product-related impurities), and comparison of samples
• Characterisation of glycosylation patterns, and variation between samples
• Integrity of protein folding, sample-to-sample variation in folding and stability against denaturation.
• Aggregation status and stability against aggregation
• Analysis of protein-protein and protein-ligand interactions

To fully apply this work we will need to develop sophisticated statistical techniques to compare large and complex data sets, to objectively compare information which previously had been inspected visually.