Method and System for Separating Biomolecules

Abstract

A separation system, and an elution arrangement to be provided in a separation system, for separating a biomolecule from a cell culture are provided.

1 . A separation system for separating a biomolecule from a cell culture, wherein said system comprises:
 a magnetic separator comprising an inlet for receiving a feed from the cell culture comprising said biomolecule and for receiving magnetic beads comprising ligands capable of binding this biomolecule, said magnetic separator being configured for separating said magnetic beads with said bound biomolecule from the rest of the feed; and   an elution arrangement comprising an elution cell, said elution cell comprising an elution cell inlet in connection with an outlet from the magnetic separator for receiving said separated magnetic beads as a slurry with buffer from the magnetic separator, wherein said elution arrangement is configured for eluting the biomolecule from the magnetic beads.   
     
     
         2 . The separation system according to  claim 1 , wherein said elution cell further comprises an elution cell outlet configured for forwarding the magnetic beads from the elution cell for reuse in the magnetic separator. 
     
     
         3 . The separation system according to  claim 1 , further comprising:
 a capturing cell connected to the inlet of the magnetic separator and comprising a cell culture inlet for receiving a feed from a cell culture and at least one magnetic bead inlet for receiving magnetic beads, said capturing cell being configured for mixing the feed from the cell culture and the magnetic beads thus allowing the specific biomolecule to bind to the magnetic beads before forwarding it to the magnetic separator.   
     
     
         4 . The separation system according to  claim 1 , wherein the magnetic separator further comprises parts of magnetic material inside the magnetic separator which parts attract the magnetic beads when a magnetic field is applied, and said magnetic separator being configured for releasing the magnetic field when the magnetic beads are to be forwarded to the elution cell, said magnetic separator further comprising a washing arrangement configured for washing out other components from the magnetic separator than those magnetically bound to the parts of magnetic material. 
     
     
         5 . The separation system according to  claim 4 , wherein said washing arrangement comprises at least one wash buffer providing arrangement connected to a pump and to the inlet of the magnetic separator possibly via a capturing cell and a wash buffer collection arrangement connected to the outlet of the magnetic separator, said washing arrangement being configured for flowing washing buffer through the magnetic separator for washing out other components of the feed than those bound to the magnetic parts. 
     
     
         6 . The system according to  claim 1 , wherein said elution arrangement further comprises a buffer providing arrangement connected to an elution cell inlet and a collection arrangement connected to an elution cell outlet and wherein said elution arrangement is configured for performing elution by providing elution buffer from the buffer providing arrangement and collecting eluate in the collection arrangement and possibly also performing cleaning in place, CIP, by providing cleaning buffer from the buffer providing arrangement and collect waste in the collection arrangement and possible also performing equilibration of the magnetic beads in the elution cell by providing equilibration buffer from the buffer providing arrangement. 
     
     
         7 . The separation system according to  claim 1 , wherein a new portion of feed from the cell culture and magnetic beads are provided into the magnetic separator while a previous portion is in the elution cell, whereby at least two portions of magnetic beads are circulating in the separation system. 
     
     
         8 . The separation system according to  claim 7 , wherein a new portion of feed from the cell culture and magnetic beads are provided into the magnetic separator while one previous portion is in the elution cell and one previous portion is in a capturing cell, whereby three portions of magnetic beads are circulating in the separation system. 
     
     
         9 . The separation system according to  claim 1 , wherein the cell culture, the magnetic separator and the elution arrangement are connected by pre-sterilized, flexible tubing and aseptic connectors, closed system. 
     
     
         10 . The separation system according to  claim 1 , wherein the elution cell further comprises an elution cell first outlet for forwarding the magnetic beads for reuse in the magnetic separator and an elution cell second outlet for collecting eluate and waste in a collection arrangement. 
     
     
         11 . The separation system according to  claim 1 , wherein the elution cell further comprises an elution cell first inlet for receiving magnetic beads from the magnetic separator and an elution cell second inlet for receiving elution buffer, cleaning in place, CIP, buffer and equilibration buffer from a buffer providing arrangement. 
     
     
         12 . The separation system according to  claim 1 , wherein said elution cell further comprises a retaining arrangement for keeping the magnetic beads within the elution cell and allowing excess buffer to escape from the elution cell. 
     
     
         13 . The separation system according to  claim 13 , wherein said retaining arrangement is a pinch, a magnetic force or a filter. 
     
     
         14 . The separation system according to  claim 1 , wherein the elution cell further comprises an adaptor for packing the magnetic beads in the elution cell and a retaining arrangement configured for keeping the magnetic beads inside the elution cell while allowing other buffer to escape from the elution cell. 
     
     
         15 . The separation system according to  claim 14 , wherein the adaptor further is configured to be used for forwarding the magnetic beads for reuse in the magnetic separator by pushing the adaptor towards an outlet of the elution cell and removing a retaining arrangement configured for keeping the magnetic beads inside the elution cell. 
     
     
         16 . The separation system according to  claim 1 , wherein the elution cell further comprises an adaptor which is positioned at a start positon in the elution cell when a slurry of magnetic beads starts to fill the elution cell from a bottom inlet of the elution cell, said start position being closer to the bottom inlet than an end position, which end position corresponds to the position of the adaptor when the filling of magnetic beads into the elution cell is completed, said adaptor keeping a force towards the flow direction of the slurry and allowing buffer to escape though the adaptor during the filling. 
     
     
         17 . The separation system according to  claim 1 , further comprising an intermediate cell connected to an elution cell outlet and configured for receiving the magnetic beads form the elution cell, said intermediate cell further being configured for forwarding the magnetic beads for possible reuse in the magnetic separator. 
     
     
         18 . The separation system according to  claim 17 , wherein said intermediate cell comprises a draining arrangement for removing excess buffer from the intermediate cell. 
     
     
         19 . The separation system according to  claim 18 , wherein the draining arrangement comprises a magnet for keeping the magnetic beads inside the intermediate cell by magnetic force while draining the intermediate cell from buffer. 
     
     
         20 . An elution arrangement configured for being connected in a separation system according to  claim 1 , said elution arrangement comprising an elution cell, said elution cell comprising an elution cell inlet configured for being connected to an outlet from a magnetic separator for receiving magnetic beads as a slurry with buffer from the magnetic separator, wherein said elution arrangement is configured for eluting a biomolecule from the magnetic beads. 
     
     
         21 . The elution arrangement according to  claim 20 , wherein said elution cell further comprises an elution cell outlet configured for forwarding the magnetic beads from the elution cell for reuse in the magnetic separator. 
     
     
         22 . The elution arrangement according to  claim 20 , wherein said elution arrangement further comprises a buffer providing arrangement connected to an elution cell inlet and a collection arrangement connected to an elution cell outlet and wherein said elution arrangement is configured for performing elution by providing elution buffer from the buffer providing arrangement and collecting eluate in the collection arrangement and also performing cleaning in place, CIP, by providing cleaning buffer from the buffer providing arrangement and collect waste in the collection arrangement and also performing equilibration of the magnetic beads in the elution cell by providing equilibration buffer from the buffer providing arrangement. 
     
     
         23 . The elution arrangement according to  claim 20 , wherein the elution cell further comprises an elution cell first outlet for forwarding the magnetic beads for reuse in the magnetic separator and an elution cell second outlet for collecting eluate and waste in a collection arrangement. 
     
     
         24 . The elution arrangement according to  claim 20 , wherein the elution cell further comprises an elution cell first inlet for receiving magnetic beads from the magnetic separator and an elution cell second inlet for receiving elution buffer, cleaning in place, CIP, buffer and equilibration buffer from a buffer providing arrangement. 
     
     
         25 . The elution arrangement according to  claim 20 , wherein said elution cell further comprises a retaining arrangement for keeping the magnetic beads within the elution cell and allowing excess buffer to escape from the elution cell. 
     
     
         26 . The elution arrangement according to  claim 25 , wherein said retaining arrangement is a pinch, a magnetic force or a filter. 
     
     
         27 . The elution arrangement according to  claim 20 , wherein the elution cell further comprises an adaptor for packing the magnetic beads in the elution cell and a retaining arrangement configured for keeping the magnetic beads inside the elution cell while allowing other buffer to escape from the elution cell. 
     
     
         28 . The elution arrangement according to  claim 27 , wherein the adaptor further is configured to be used for forwarding the magnetic beads for reuse in the magnetic separator by pushing the adaptor towards an outlet of the elution cell and removing a retaining arrangement configured for keeping the magnetic beads inside the elution cell. 
     
     
         29 . The elution arrangement according to  claim 20 , wherein the elution cell further comprises an adaptor which is positioned at a start positon in the elution cell when a slurry of magnetic beads starts to fill the elution cell from a bottom inlet of the elution cell, said start position being closer to the bottom inlet than an end position, which end position corresponds to the position of the adaptor when the filling of magnetic beads into the elution cell is completed, said adaptor keeping a force towards the flow direction of the slurry and allowing buffer to escape though the adaptor during the filling.