IP Library Granted Patent US 7,192,767
Granted Patent B2
US 7,192,767 · App. 10/221,124 · Granted Mar 20, 2007

Matrix reactor and a method for producing products in said reactor

View Patent ↗
Loading inventors, assignments & file history…
Monitor This Case
Get email alerts when status or documents change.
Order Certified Copies
Most orders are placed with the USPTO same day — all within 24 business hours.
Order via The Patent Place →
Pre-filled with this patent's details
Quick Facts
Patent No.
US 7,192,767
App. No.
10/221,124
Granted
Mar 20, 2007
Kind
B2
Abstract

The present invention concerns a matrix reactor for reactions in which low-molecular and high-molecular reactants are involved wherein the reactor space V R contains a matrix and comprises a reaction compartment and a supply compartment, wherein the matrix is composed of a porous material which can take up low-molecular reactants and wherein the exclusion volume V 0 of the matrix is available as the reaction compartment and the matrix volume V M of the matrix is available as the supply compartment. The matrix reactor can be used for enzymatic processes, in particular for a coupled in vitro transcription/translation reaction to produce proteins.

Claims (32)

1. A process for carrying out a coupled in vitro transcription and translation reaction to produce proteins, said method comprising the steps of:

providing a reactor, said reactor comprising walls that define a total reactor volume V R , a matrix comprising a porous material held within said total reactor volume, the matrix dividing the total reactor volume into a matrix volume V M and a matrix exclusion volume V 0 , the division based on the size of the matrix pores and in the absence of a membrane separating the matrix volume V M and the matrix exclusion volume V 0 , wherein at least 20 vol % of the total reactor volume is filled with said matrix,

contacting said reactor with a supply solution, said supply solution comprising each of the 20 naturally occurring amino acids, ATP, GTP, CTP, and UTP, wherein said components of the supply solution have a molecular weight smaller than the pores of said matrix and are taken up into the matrix volume V M and are distributed largely homogeneously in the total reactor volume V R ,

contacting said reactor with a reaction solution, said reaction solution comprising tRNAs, DNA templates, RNA polymerases, mRNA, and enzymes, wherein said tRNAs, DNA templates, RNA polymerases, mRNA, and enzymes each have a molecular weight larger than the pores of said matrix and are only localized in the matrix exclusion volume V 0 ,

moving the reaction solution through the matrix exclusion volume V 0 while the reactor is thermostatted at a temperature conducive to in vitro transcription and translation, and

obtaining said protein by recovering an eluate from the reactor.

2. The process as claimed in claim 1 , wherein

the matrix is first equilibrated with the supply solution, before

the equilibrated matrix in the reactor volume is contacted with the reaction solution and wherein

the reactor is maintained at a constant temperature.

3. The process as claimed in claim 2 , wherein

the reaction solution is applied to the column and

the reaction solution is held in the column for the duration of the reaction.

4. The process as claimed in claim 2 , wherein

the reaction solution is applied to the column and

additional reaction solution is pumped continuously or step-wise through the column for the duration of the reaction.

5. The process as claimed in claim 2 , wherein

the reactor is a batch reactor and

the reaction solution and equilibrated matrix are brought together in the reactor volume,

the added matrix volume being at least 20 vol % of the volume of the reactor volume.

6. The process as claimed in claim 2 , wherein

the reactor is a continuous exchange cell-free reactor having a reaction compartment,

the matrix is incorporated into the reaction compartment of the continuous exchange cell-free reactor and

the matrix volume is more than 20 vol % of the volume of the reaction compartment.

7. The process as claimed in claim 2 , wherein

the reactor is a continuous flow cell-free reactor having a reaction compartment,

the matrix is incorporated into the reaction compartment of the continuous flow cell-free reactor and

the matrix volume is more than 20 vol % of the volume of the reaction compartment.

8. The process of claim 2 wherein said matrix comprises particles having a particle size in the range of 0.1 to 1000 μm.

9. The process of claim 8 wherein said matrix has an exclusion limit selected from a range of about 10 to about 100 kDa.

10. The process of claim 9 wherein about 80 to about 100 vol % of the total reactor volume is filled with said matrix.

11. The process of claim 8 wherein said matrix comprises a mixture of particles having different exclusion ranges independently selected from the range of about 10 to about 100 kDa.