ABSTRACT
This chapter aims to aid in the design and operation of large-scale conventional chromatography for the purification of proteins and peptides. For small-volume high-value products containing closely related impurities, large-scale high-performance liquid chromatograph should be considered if resolution by conventional chromatography is inadequate. On-off chromatography accounts for almost all of protein and large-scale peptide chromatography. Frontal chromatography, also known as flow-through or breakthrough chromatography, is a mode in which conditions are chosen such that the desired component very quickly saturates the column and emerges from the column shortly after the feed is started. The lab protocol should be developed to the point that the behavior of the chromatography can be predicted reliably. Frontal chromatography should be considered in every case to see if conditions can be found such that the impurities are retained while the product passes through the column.
