Patent ID: 9149738
Filing Date: 2015-10-06
Classification: B01D,G01N

Abstract:
1. A method of purifying target molecules from one or more impurities in a sample, the method comprising the steps of: a) providing at least three separation units having the same chromatography matrix which are connected so that liquid can at least flow from one separation unit to the subsequent one and from the last to the first separation unit, so that the at least three separation units are connected for a circle of flow between them; b) feeding the sample to a first separation unit wherein the sample is at a first pH and conductivity enabling the target molecules to be bound to the chromatography matrix in this separation unit, said separation unit is, for at least part of the loading time, in fluid communication with the next separation unit in the circle so that target molecules not bound to the chromatography matrix in said first separation unit bind to the chromatography matrix in the next separation unit, and, at the same time as said feeding steps, at least washing, eluting and re-equilibrating one separation unit different from the separation unit that is being loaded and from the one that is in fluid communication with the separation unit that is being loaded; c) switching the feed to the next separation unit; d) feeding the sample on the next separation unit wherein the sample is at a pH and conductivity enabling the target molecules to be bound to the chromatography matrix in said next separation unit, said next separation unit is, for at least part of the loading time, in fluid communication with the separation unit after the next one in the circle so that target molecules not bound to said next separation unit bind to the chromatography matrix in the separation unit after the next, and, at the same time at least washing, eluting and/or re-equilibrating one separation unit different from the separation unit that is being loaded and from the one that is in fluid communication with the separation unit that is being loaded; e) repeating steps c) and d) one or more times; wherein the feed is continuous into at least one of the separation units and has a velocity above 800 cm/h and wherein the chromatography matrix of the separation units comprises particles with a diameter between 40 and 200 μm and with pore diameters in the range between 50 nm and 200 nm.