US 7,238,522 B2Granted

Devices and methods for biomaterial production

AssigneeADVISYS, Inc.

InventorsHenry Hebel, Sriram Ramakrishnan, Hugo Gonzalez, Jeff Darnell

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Abstract

An apparatus and a method for isolating a biologic product, such as plasmid DNA, from cells. The method involves lysing cells in a controlled manner separate insoluble components from a fluid lysate containing cellular components of interest, followed by membrane chromatographic techniques to purify the cellular components of interest. The process utilizes a unique lysis apparatus, ion exchange and, optionally, hydrophobic interaction chromatography membranes in cartridge form, and ultrafiltration. The process can be applied to any biologic product extracted from a cellular source. The process uses a lysis apparatus, including a high shear, low residence-time mixer for advantageously mixing a cell suspension with a lysis solution, a hold time that denatures impurities, and an air-sparging bubble mixer that gently yet thoroughly mixes lysed cells with a neutralization/precipitation buffer and floats compacted precipitated cellular material.

Claims14

Dependent claims

An apparatus for isolating cellular component of interest from cells comprising:

(a) a first tank, wherein the first tank is used for holding a suspension of cells having the cellular component of interest;
(b) a mixer in fluid communication with the first tank;
(c) a second tank in fluid communication with the mixer, wherein the second tank is used for holding a lysis solution;
(d) a holding coil in fluid communication with the mixer;
and
(e) a bubble-mixer chamber with a top and a bottom having:
(i) a first inlet in fluid communication with the holding coil;
(ii) a second inlet in fluid communication with a third tank, wherein the third tank is used for holding a precipitation solution, a neutralization solution, or a mixture thereof;
(iii) a third inlet in fluid communication with a gas source;
(iv) a vent;
and
(v) an outlet in fluid communication with a fourth tank, wherein the fourth tank is used for separating precipitated cellular components from fluid cell lysate;
wherein,
the mixer is a high-shear, low-residence-time-mixing-device, wherein the residence-time is less than or equal to about one second;
the suspension of cells having the plasmid DNA from the first tank is allowed to flow into the mixer;
the lysis solution from the second tank is allowed to flow into the mixer;
a lysate-mixture is allowed to flow from the mixer into the holding coil;
the lysate-mixture from the holding coil is allowed to flow into the bubble-mixer chamber;
the precipitation solution, the neutralization solution, or the mixture thereof from the third tank is allowed to flow into the bubble-mixer chamber;
and a suspension containing the cellular component of interest is allowed to flow from the bubble-mixer chamber into the fourth tank.

17 more elements

Dependencies are inferred from the claim text.

Facts

Application: 10/857,439Filed 27 May 2004 · us provisional application
Priority: 30 May 2003
Published: US 2005/0014245 A120 Jan 2005
Granted: 3 Jul 2007
Term ends: Not estimatedThe source records don’t support an estimate.

Classification12 codes

C12N 15/1003
Processes for the isolation, preparation or purification of DNA or RNA: extracting or separating nucleic acids from biological samples, e.g. pure…
B01F 33/406
Mixers using gas or liquid agitation, e.g. with air supply tubes: in receptacles with gas supply only at the bottom
C12M 47/06
Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass: hydrolysis; Cell lysis;…
C12M 47/10
Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass: separation or…