Purification method and uses thereof

A method according to any of the preceding claims, wherein before for the first time carrying out the first interconnected step a start-up step (BSU) is carried out, in which during a batch timespan (tBSU) said adsorber sections (1,2) are disconnected and an equilibrated adsorber section (1) to be taking the place of the first adsorber section (1) in the subsequent first interconnected step is loaded with feed mixture via the first adsorber section inlet and product is collected from first adsorber section outlet, while the second adsorber section (2) is either being equilibrated or already equilibrated and inactive;

A method according to any of the preceding claims, wherein after termination of the desired cycles of steps a.-f. the procedure is followed by a shut-down sequence (BSD), wherein during a first batch timespan (tBSDa) said adsorber sections (1,2) are disconnected and the adsorber section having been subjected to washing in the preceding second batch step (B2) is washed such that product is collected from the adsorber section outlet;

• and wherein during a second timespan (tBSDb) said adsorber section is regenerated;

A method according to any of the preceding claims, wherein the chromatographic stationary phase in the adsorbers is in the form of particles, such as beads, or of membranes or monoliths or fibers and wherein the adsorbers are single-use or multi-use devices, and wherein preferably the chromatographic stationary phase in the adsorbers is a non-affinity material with a propensity to withhold the product and the impurities depending on their chemical nature in a gradual manner.

A method according to any of the preceding claims wherein the chromatographic adsorption is based on hydrophobic interaction, multi-modal interaction or ion exchange.

A method according to any of the preceding claims, wherein a detector is located at the outlet of each adsorber section and wherein the changes in the detected signal over time during the first and second interconnected phases (IC1a, IC1b, IC2a, IC2b) are used to determine the first and second interconnected timespans (tICa, tICb), respectively.

A method according to any of the preceding claims, wherein the product is one or a group of chemical reaction products, chemical separation products, biochemical reaction products, biological products, wherein preferably the reaction products are natural products, metals, antibodies, antibody fragments, fusion proteins, recombinant glycoproteins, and/or plasma proteins, or derivatives and/or combinations and/or mixtures thereof.

A method according to any of the preceding claims, wherein the product is an antibody monomer and the impurities include at least one antibody dimer and/or multimer.

A method according to any of the preceding claims, wherein in the second interconnected step (IC1b) the stream exiting the first adsorber section outlet is diluted inline before entering the second adsorber section inlet, and wherein in the first batch step the second adsorber section inlet is loaded with a second washing buffer, and wherein preferably in the fourth interconnected step (IC2b) the stream exiting the second adsorber section outlet is diluted inline before entering the first adsorber section inlet, and wherein in the first batch step the first adsorber section inlet is loaded with a second washing buffer.

A method according to any of the preceding claims, wherein in the second interconnected step (IC1b) the stream exiting the first adsorber section outlet is diluted inline before entering the second adsorber section inlet, and wherein in the first batch step the second adsorber section inlet is loaded with feed mixture, and wherein preferably in the fourth interconnected step (IC2b) the stream exiting the second adsorber section outlet is diluted inline before entering the first adsorber section inlet, and wherein in the first batch step the first adsorber section inlet is loaded with feed mixture.

A method according to any of the preceding claims, wherein in the second interconnected step (IC1b) the stream exiting the first adsorber section outlet is supplemented with feed mixture before entering the second adsorber section inlet, and wherein in the first batch step the second adsorber section inlet is loaded with feed mixture, and wherein preferably in the fourth interconnected step IC2b the stream exiting the second adsorber section outlet is supplemented with feed mixture before entering the first adsorber section inlet, and wherein in the first batch step the first adsorber section inlet is loaded with feed mixture.

A method according to any of the preceding claims, wherein in the second interconnected step (ICb) the stream exiting the first adsorber section outlet is diluted inline before entering the second adsorber section inlet with a third washing buffer, and wherein preferably in the fourth interconnected step IC2b the stream exiting the second adsorber section outlet is diluted inline before entering the first adsorber section inlet with a third washing buffer.

Use of a method according to any of the preceding claims for the purification of monoclonal antibodies, in particular for the separation of monomers from dimers and/or multimers.