Example: Peptide Synthesis Optimization via EasyMax & DoE
Using a peptide synthesis from LONZA, the objective of this work was to:
- Identify the reaction parameters necessary to create a kinetic model
- Maximize yield and selectivity
- Minimize by-product formation.
Four parameters were chosen related to concentration of reagents and temperature, and a DoE consisting of 11 experiments were run in parallel in the EasyMax reactors.
HPLC analysis showed that at a temperature of 10.0 °C, the conversion and yield after 90 minutes was at a minimum level, as expected. However, the level of impurities was also minimized. Increasing temperature improved conversion and yield, but this caused a loss of selectivity and more impurities. Using a lower temperature and lower amount of one of the reagents (EDC) reduced the impurities. In contrast, low TEA concentrations resulted in higher impurities.
The combination of statistical experimentation using parallel synthesis reactors accelerated the optimization of this peptide synthesis.
Urs Groth, Fabio Visentin, “Fast and Effective Synthesis of Peptides: A Statistical Approach Using EasyMax”, Mettler Toledo Application Note 2010-002