A novel NMR-based method for quality assurance of chiral purity of peptides in clinical development and production

Purpose and goal

The aim of the project was to develop a competitive method for chiral analysis of peptides. The method is based on NMR and the application of chiral shift reagents for separation of amino acid L- and D-enantiomers. Using model systems, the project has visualized possibilities and challenges regarding applicability, sensibility and reproducibility of the method, along with estimated duration and cost efficiency. A standardized method for chiral analysis of amino acid mixtures is, however, not yet implemented.

Results and expected effects

The project has managed to survey the potential of the method to become commercially viable, from both a technical and economical/market oriented perspective. A number of technical challenges have been identified. One major challenge is the development of a cost efficient NMR shift reagent for chiral analysis of complex amino acid mixtures. The project has also generated results that unsuspectedly provides insight into strengths and drawbacks in the currently controlling method on the market.

Approach and implementation

1.Literature study of published methods for chiral separation of amino acids with NMR shift reagents. 2.Optimization of model system for chiral separation of L- and D-enantiomers. Study of reproducibility and limit of quantification. 3.Hydrolysis of model peptide and NMR study of hydrolysate. 4.Evaluation of chiral shift reagent for NMR samples of hydrolysate. 5.Development of quality system including a quantitative NMR method. All steps have been performed. Developing work remains for step 2-4.