US 12,607,626 B2Granted

Method for separating biomolecules

AssigneeCytiva Sweden AB

InventorsOla Lind, Nils Norrman, Sara Häggblad Sahlberg, Ronnie Palmgren

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Abstract

The present disclosure is directed to a method for separating a biomolecule from a cell culture or from a biological solution, comprising the steps of: (a) providing magnetic particles comprising ligands capable of binding the biomolecule; (b) contacting a cell culture or a biological solution comprising the biomolecule with the magnetic particles to obtain magnetic particles comprising the bound biomolecule; (c) retaining the magnetic particles with a magnetic field in a magnetic separator; (d) optionally washing the magnetic particles with a washing liquid; (e1) providing a flow of an elution liquid through the magnetic separator to elute the bound biomolecule from the magnetic particles while retaining the magnetic particles with the magnetic field in the magnetic separator; (f1) forwarding the biomolecule eluted from the magnetic separator to a membrane chromatography device; (g1) separating the biomolecule from impurities and/or contaminants by membrane chromatography.

Claims23

Dependent claims

A method for separating a biomolecule from a cell culture, comprising the steps of:

(a) providing magnetic particles comprising ligands capable of binding the biomolecule;
(b) contacting a cell culture comprising the biomolecule with the magnetic particles to obtain magnetic particles comprising the bound biomolecule;
(c) retaining the magnetic particles with a magnetic field in a magnetic separator;
(d) washing the magnetic particles at least once with a washing liquid in continuous mode, wherein the wash comprises the following substeps:
(d1) agitating the magnetic particles in at least one plane of the magnetic separator by applying an oscillating magnetic field to form a fluidised bed of magnetic particles in the magnetic separator, and
(d2) providing a flow of a washing liquid in a direction essentially perpendicular to the at least one plane to remove the cell culture while retaining the magnetic particles in the magnetic field, wherein the flow of washing liquid comprises simultaneous introduction and removal of washing liquid;
(e) agitating the magnetic particles in at least one plane of the magnetic separator by applying an oscillating magnetic field to form a fluidised bed of magnetic particles in the magnetic separator;
and
(f) providing a flow of an elution liquid in a direction essentially perpendicular to the at least one plane, to elute the bound biomolecule from the magnetic particles while retaining the magnetic particles with the magnetic field in the magnetic separator,
wherein steps (c) and (d) are performed simultaneously.

7 more elements

Dependencies are inferred from the claim text.