Cation chromatography using predicted elution buffer salt concentration

A technology of elution buffer and salt concentration, applied in the field of chromatography and cation chromatography

Pending Publication Date: 2022-04-12
F HOFFMANN LA ROCHE & CO AG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] Thus, when it comes to selectively eluting a protein with a specific PTM, or when it comes to obtaining an elution comprising a desired ratio of two or more protein variants that differ from each other only in the type, number and / or location of the PTM Problems with current chromatographic techniques for separating protein variants that undergo glycosylation and / or other forms of PTMs are serious problems when

Method used

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  • Cation chromatography using predicted elution buffer salt concentration
  • Cation chromatography using predicted elution buffer salt concentration
  • Cation chromatography using predicted elution buffer salt concentration

Examples

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Embodiment Construction

[0219] figure 1 A flow diagram of a chromatography method according to an embodiment of the invention is depicted.

[0220] This method can be used to obtain a target elution volume comprising a desired absolute or relative amount and / or purity of two or more target proteins. For example, the method may be performed to obtain target protein P1 and target protein P2 respectively in a certain concentration or amount, wherein target protein P1 preferably has the lowest purity within the target elution volume. Additionally or alternatively, the method can be used to obtain a target elution volume comprising a desired ratio of two or more target proteins.

[0221] In a first step 100, the method includes providing, for example, figure 2 and image 3 The cation exchange column depicted in 230. There are a large number of commercially available chromatography resin materials that can be used to manufacture self-packed as well as prepacked columns.

[0222] For example, Poros ...

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Abstract

The present invention relates to a chromatographic method for generating a target elution volume (238) comprising a first target protein and a second target protein (P1, P2). The method comprises: providing (102) a cation exchange chromatography column (230); applying (104) a protein solution (202) on the column, the protein solution comprising the first target protein (P1), the second target protein, and optionally one or more additional proteins (P3, P4, P5, P6); inputting (106) an optimization criterion; calculating (108) a chromatographic simulation for calculating an elution buffer salt concentration (222), the elution buffer salt concentration being adapted to provide a target elution volume that optimally matches the optimization criteria; calculating (110) a collection boundary of the target elution volume from at least the calculated salt concentration and an input optimization criterion; applying (112) an elution buffer (226) having the calculated salt concentration on the chromatography column; performing (114) the elution; and collecting (116) the calculated target elution volume.

Description

technical field [0001] The present invention relates to the field of chromatography, and in particular to the field of cation chromatography for obtaining specific proteins. Background technique [0002] Cation exchange chromatography is a form of ion exchange chromatography commonly used to separate molecules based on their net surface charge. Negatively charged ion exchange resins that have an affinity for molecules with a net positive surface charge, such as proteins or peptides, are used for both preparative and analytical purposes. [0003] The net surface charge of a protein varies with pH in a manner determined by the protein's isoelectric point ("pi"). Proteins with different pI values ​​will have different degrees of charge at a given pH and thus have different affinities for the negatively charged surface groups on the ion exchange media particles in the column. Therefore, proteins with different pI values ​​will bind to the chromatographic resin with different s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/28C07K1/18B01D15/36
CPCG16C20/30B01D15/362B01J47/02G16C20/10B01D15/166B01J39/07C07K1/18G01N30/64G01N2030/027G01N2030/067
Inventor A·肖布马尔F·维特科普
Owner F HOFFMANN LA ROCHE & CO AG
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