In this presentation, Arnaud Delobel explains how the development of liquid chromatography (LC) methods has long been done using a trial-and-error approach, also known as “one factor at a time” (OFAT). While this approach is the easiest to implement, it is both time-consuming and quite inefficient. Usually, this approach does not lead to the optimal method in terms of performance and robustness.
Instead of the usual approach, retention modelling is the method of choice for LC method development:
- There is no need to know the structure of the solutes to be separated.
- The retention behaviour of common solutes, including proteins, is well known in many HPLC modes.
- Prediction is based on well-established empirical models.
- By running a few initial experiments (between 2 and 12), the separation can be predicted to any conditions.
- Changes in conditions can be studied in silico (for example, change of column dimensions, flow rate, gradient time, etc.).
This approach is very valuable for the separation of proteins. Platform methods are often used, but it is often useful to have a product-specific method for improved performance. With a computer-based approach, the optimisation can be straightforward.
Download the slides of the presentation below to discover how software-assisted method development (using DryLab) may be a powerful tool for efficient development of LC separations of your biomolecules.