Systems and methods for preparing a polypeptide from a mixture

A method for preparing a target polypeptide from a mixture including the target polypeptide, the method comprising:

• passing the mixture including the target polypeptide to a first column of a plurality of chromatographic columns in a hydrophobic interaction chromatography (HIC) apparatus, wherein each of the plurality of columns comprises an outlet connectable to another column of the plurality of columns;
• passing an effluent including the target polypeptide from the first column to a second column of the plurality of columns;
• passing one or more mobile phases to a third column of the plurality of columns;
• and
• passing the target polypeptide through the outlets of each of the plurality of columns, wherein a sum of residence times for the mixture including the target polypeptide in the first column and second column is substantially the same as the sum of the residence times of the one or more mobile phases in the third column.

A method for preparing an antibody using a plurality of chromatographic columns, wherein each of the plurality of chromatographic columns includes a hydrophobic interaction medium, the method comprising:

• in a first stage:
• loading a quantity of a mixture including the antibody into a first column of the plurality of columns;
• loading a quantity of the mixture into a second column of the plurality of columns via the first column;
• and
• performing a non-loading step including at least one of washing, stripping, and equilibration processes in a third column of the plurality of columns;
• in a second stage:
• loading a quantity of the mixture including the antibody into the second column;
• loading a quantity of the mixture into the third column via the second column;
• and
• performing the non-loading step comprising at least one of washing, stripping, and equilibration processes in the first column;
• and
• in a third stage:
• loading a quantity of the mixture including the antibody into the third column;
• loading a quantity of the mixture into the third column via the second column;
• and
• performing the non-loading step including at least one of washing, stripping, and equilibration processes in the second column.