Liquid Chromatography
Strategic Peptide Purification with YMC
Oct 22 2024
The broad functional spectrum of peptides opens up numerous opportunities for pharmaceutical applications. Reversed-phase chromatography has established itself as the preferred purification method. With the right stationary phase, structural variants and even peptides with minimal sequence differences can be precisely separated.
The new poster from YMC focuses on maximising yield, reducing process times, and ensuring economic feasibility. In general, the most comprehensive approach for method development in preparative LC is the so-called linear scale-up, which consists of 3 steps:
Step 1: Method development at analytical scale
The first step is the identification of an optimal stationary phase ligand. This mainly depends on the content of hydrophobic amino side chains, potential secondary structures of the peptide and additional modifications. The innovative RP stationary phases YMC-Triart Prep which are the first choice for peptide purifications are based on a hybrid-silica base and are available with different modifications from C4 to C18 covering all relevant ligands.
In addition, the right pore and particle size have to be selected. Generally, the pore size should be as small as possible but large enough for separation efficiency. The particle size of the stationary phase determines the resolution and also the productivity of the overall process. With various combinations of pore and particle sizes, YMC-Triart Prep allows full flexibility in method development and therefore leads to tailored solutions for all types of purification processes.
Another advantage: YMC-Triart Prep provides high chemical stability even at elevated pH. Due to this stability, purifications at alkaline pH are possible. This is advantageous for many peptide purifications and drastically increases the possible method set-ups. An additional benefit is the Cleaning-in-Place (CIP) stability of YMC-Triart Prep which restores the column performance and increases the column lifetime.
Step 2: Loadability studies at analytical scale
Loadability studies at analytical scale are the next step towards a highly productive preparative scale purification. The loadability depends on the properties of the stationary phase as well as on the peptide itself and the impurity profile. Due to the optimised particle technology of YMC-Triart Prep, high sample loadings are possible.
Step 3: Scale-up to final process scale
The final step in process development is the scale-up to preparative or even industrial scale. At this point all relevant parameters were optimized leading to a final purification method that can be scaled up to large scale. For detailed information regarding preparative scale-up, please refer to the YMC Technical Note on linear scale-up.
Conclusion
This efficient yet comprehensive poster makes it easy to achieve the goal of cost-effective purification of a target peptide. The success of this development strategy is impressively demonstrated on the poster using a real peptide purification process!
In addition to the right strategy for method development, the right choice of stationary phase also plays an important role in establishing a productive and efficient purification process. The innovative hybrid-silica YMC-Triart Prep is highly chemically and mechanically stable. Together with the long lifetime, this directly leads to an increased productivity. YMC-Triart Prep is available in large scale quantities for a safe and reliable supply.
For more information visit our website or get in touch with your local YMC representative.
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