A critical and fundamental factor in both analytical and preparative chromatography is the use of true buffers. In other words, mobile phases with buffering capacity. For example, in peptide purification, there are concentrated bands of various components of the sample, including the target peptide, moving through the column and you need to have control over their tendency to dissociate...
Peptide therapeutic agents are enjoying increasing success but they are challenging to produce and, not least, to purify. Purity requirements are, in fact, approaching those for small molecules. Synthesis generates a crude mixture containing failed sequences and chemical variants.
The global diabetes pandemic has created a massive demand for insulin, especially in China, which is expected to soon have half the world’s diabetes patients. Meeting this demand means developing extremely efficient production methods that can supply affordable insulin with high purity. A promising approach is to add an orthogonal step upstream the main reverse-phase chromatography (RPC) peptide purification to increase purity and also guard the expensive high-performance RPC columns from bioburden and stringent cleaning-in-place procedures.
The purity requirements for therapeutic peptides are very stringent, but synthesis generates a crude peptide mixture containing failed sequences and chemical variants, while recombinant peptides have a considerable bioburden from the host cell. These crude feeds can foul the reverse phase chromatography (RPC) columns commonly used in peptide purification. Introducing an orthogonal purification step upstream significantly reduces the burden on the expensive high performance RPC column, increases the peptide yield and purity, and also prolongs column lifetime.