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US 11,590,434 B2Granted
Method and apparatus for chromatographic purification
AssigneeMerck Patent GmbH
InventorRomas Skudas
Life of this patent
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Abstract
A method and an apparatus suitable for a continuous chromatography process which only needs three separation columns, and a two-step process containing two chromatographic steps, in which the first chromatographic step (capture) is performed alternating and sequentially on two separation columns, the second chromatographic step (polishing) is performed, also sequentially, on the third column.
Claims6
A method of purifying a target molecule from one or more impurities in only one sample using an apparatus comprising
- two separation units A1 and A2 both having the same chromatography matrix and a separation unit B having a chromatography matrix which differs from the chromatography matrix of separation units A1 and A2, all separation units having a fluid inlet and a fluid outlet, whereby there is at least fluid connection between the fluid outlet of separation unit A1 and the fluid inlet of separation unit B and fluid connection between the fluid outlet of separation unit A2 and the fluid inlet of separation unit B
- at least one valve in the fluid connection between separation units A1 and A2 and separation unit B that allows to switch between fluid communication between the outlet of separation unit A1 and the fluid inlet of separation unit B and fluid communication between the outlet of separation unit A2 and the fluid inlet of separation unit B,
- at least two buffer reservoirs and at least two pumps
- a reservoir containing sample solution that is in fluid connection with the inlets of separation units A1 and A2,
- the method comprising:
- alternately loading portions of the sample on the separation units A1 and A2 so that while the sample is loaded on separation unit A1 wherein the sample is at a first pH and conductivity enabling the target molecule to be bound to separation unit A1, separation unit A2 is at least part of that time in fluid communication with separation unit B so that the target molecule loaded on separation unit A2 is eluted onto separation unit B and separation unit A2 is re-equilibrated and while the sample is loaded on separation unit A2 wherein the sample is at a first pH and conductivity enabling the target molecule to be bound to separation unit A2, separation unit A1 is at least part of that time in fluid communication with separation unit B so that the target molecule loaded on separation unit A1 is eluted onto separation unit B and separation unit A1 is re-equilibrated,
- recovering the target molecule from the fluid outlet of separation unit B.
4 more elements
Dependencies are inferred from the claim text.