Filter based system for capture of lysis of suspended cells

Patent metadata

Field	Value
Publication number	US-10364414-B2
Application number	US-201715700474-A
Country	US
Kind code	B2
Filing date	Sep 11, 2017
Priority date	Oct 23, 2015
Publication date	Jul 30, 2019
Grant date	Jul 30, 2019

Abstract

A cell lysis workflow involving a cell suspension that is passed through a filter in a first flow direction from a first side of the filter toward a second side thereof. The filter captures a plurality of cells on the first side. A lysis solution is passed through the filter in a second flow direction opposite the first direction, thereby dislodging the plurality of cells captured on the filter, and resuspending the plurality of cells in the lysis solution on the first side of the filter. The lysis solution lyses the plurality of cells to produce a cellular lysate. The cellular lysate is passed through the filter in the first flow direction.

First claim

What is claimed is:

1. A cell lysis system comprising:
a filter;
a reservoir in fluid communication with the filter and operatively positioned on a first side thereof;
a cell suspension intake in fluid communication with the filter and operatively positioned on the first side thereof;
a lysis solution intake in fluid communication with the filter and operatively positioned on a second side thereof; and
a reversible pump in fluid communication with the filter, wherein the reversible pump has a first operating state to cause fluid flow in a first flow direction through the filter from the first side toward the second side thereof, and a second operating state to cause fluid flow in a second flow direction through the filter from the second side toward the first side thereof;
wherein the cell suspension intake is configured to receive a cell suspension therethrough such that the cell suspension is filtered through the filter and a plurality of cells of the cell suspension are captured on the first side of the filter when the reversible pump is in the first operating state;
wherein the lysis solution intake is configured to receive a lysis solution therethrough such that the lysis solution passes through the filter in the second flow direction when the reversible pump is in the second operating state to lyse and suspend the plurality of cells captured on the first side of the filter to produce a cellular lysate; and
wherein the reservoir is configured to receive the cellular lysate when the reversible pump is in the second operating state.

2. The cell lysis system of claim 1 , wherein the filter is a hollow fiber filter.

3. The cell lysis system of claim 1 , wherein the reservoir and the filter are integrated into a unitary housing.

4. The cell lysis system of claim 1 , further comprising a purification column to isolate a protein from the cellular lysate, wherein the purification column is in fluid communication with the second side of the filter, and wherein the purification column is configured to receive the cellular lysate from the reservoir when the reversible pump is in the first operating state.

5. The cell lysis system of claim 1 , further comprising one or more chromatography columns to isolate or purify a protein from the cellular lysate.

6. The cell lysis system of claim 5 , wherein the chromatography column is an affinity chromatography column, a gel filtration chromatography column, an ion exchange chromatography column, or any combination thereof.

7. The cell lysis system of claim 1 , wherein the reversible pump is operable to selectively cause fluid to pass through the filter in the first flow direction or the second flow direction.

8. The cell lysis system of claim 1 , wherein the reversible pump is a peristaltic pump.

9. The cell lysis system of claim 1 , further comprising a reservoir on the first side of the filter, wherein the reservoir receives the cellular lysate.

Assignees

• Life Technologies Corp

Inventors

• Martin Philip
• Nelson Paul
• Bourdon David

Classifications

• C07K1/18

  Ion-exchange chromatography · CPC title

• C07K1/34

  by filtration, ultrafiltration or reverse osmosis · CPC title

• C12M29/04

  Filters; Permeable or porous membranes or plates, e.g. dialysis · CPC title

• C07K1/22

  Affinity chromatography or related techniques based upon selective absorption processes · CPC title

• C12M29/16

  Hollow fibers (hollow fiber modules in general B01D63/02) · CPC title