Back to results
US 10,526,574 B2Granted
Fed-batch process for the production of bacterial ghosts
AssigneeBird-C GmbH
InventorsWerner Lubitz, Christian Herwig, Timo Langemann, Patrick Sagmeister
Life of this patent
Estimating the term…
Abstract
The invention relates to a fed-batch process for the production of bacterial ghosts based on the decoupling of the lytic gene expression and actual lysis of bacterial cells.
Claims31
A method for producing a bacterial ghost preparation comprising the steps:
- (a) providing bacterial cells comprising a lytic gene encoding a protein capable of forming a tunnel structure in the bacterial cell envelope wherein the expression of the lytic gene is under operative control of a regulatable promoter,
- (b) cultivating the bacterial cells under conditions wherein the lytic gene is not expressed to a final biomass concentration of about 10-80 g dry cell weight (DCW)/L,
- (c) after step (b), subjecting the bacterial cells to conditions wherein the lytic gene is expressed by inducing the regulatable promoter but lysis of the bacterial cells is repressed and wherein the bacterial cells are subjected to an at least substantially stationary state,
- (d) after step (c), subjecting the bacterial cells to conditions wherein lysis of the bacterial cells takes place, and thereafter carrying out a further promoter induction step by inducing the regulatable promoter, and
- (e) obtaining the resulting bacterial ghosts.
2 more elements
A method for recombinantly producing a protein of interest comprising the steps:
- (a) providing bacterial cells comprising (i) a lytic gene encoding a protein capable of forming a tunnel structure in the bacterial cell envelope wherein the expression of the lytic gene is under operative control of a regulatable promoter and (ii) a gene encoding a protein of interest,
- (b) cultivating the bacterial cells under conditions wherein the lytic gene is not expressed to a final biomass concentration of about 10-80 g dry cell weight (DCW)/L,
- (c) after step (b), subjecting the bacterial cells to conditions wherein the lytic gene is expressed by inducing the regulatable promoter but lysis of the bacterial cells is repressed and wherein the bacterial cells are subjected to an at least substantially stationary state,
- (d) after step (c), subjecting the bacterial cells to conditions wherein lysis of the bacterial cells takes place, and thereafter carrying out a further promoter induction step by inducing the regulatable promoter, and
- (e) recovering and optionally purifying the protein of interest, which has been expressed during step (b) and/or step (c).
2 more elements
A method for recombinantly producing bacteriophage (Φ)X174 protein E or derivatives thereof comprising the steps:
- (a) providing bacterial cells comprising a lytic gene encoding the lytic protein E or derivatives thereof wherein the expression of the lytic gene is under operative control of a regulatable promoter,
- (b) cultivating the bacterial cells under conditions wherein the lytic gene is not expressed to a final biomass concentration of about 10-80 g dry cell weight (DCW)/L,
- (c) after step (b), subjecting the bacterial cells to stationary state conditions wherein the lytic gene is expressed by inducing the regulatable promoter but lysis of the bacterial cells is repressed,
- (d) after step (c), subjecting the bacterial cells to conditions wherein lysis of the bacterial cells takes place, and thereafter carrying out a further promoter induction step by inducing the requlatable promoter, and
- (e) recovering and optionally purifying the protein E or derivatives thereof.
2 more elements
Dependencies are inferred from the claim text.