Abstract: A host-vector system that uses the RNA-based copy number control mechanism of ColE1-type plasmids for regulating the expression of a marker gene allows for antibiotic-free selection of plasmids and is useful for production of plasmid DNA and recombinant proteins.
Abstract: A process for producing plasmid DNA E. coli cells comprises a pre-culture and fed-batch process. The culture media of the batch phase and the culture medium added during the feeding phase are chemically defined. The culture medium of the feeding phase contains a growth-limiting substrate and is added, for at least a fraction of the feeding phase, at a feeding rate that follows a pre-defined exponential function, thereby controlling the specific growth rate at a pre-defined value. The process results in high yield and homogeneity of plasmid DNA.
Type:
Application
Filed:
February 19, 2009
Publication date:
October 8, 2009
Applicant:
BOEHRINGER INGELHEIM AUSTRIA GMBH
Inventors:
Hans HUBER, Gerhard WEIGL, Wolfgang BUCHINGER
Abstract: Disclosed is a method for the production of a heterologous polypeptide of interest with a homogenous N-terminus, using a fusion polypeptide comprising the polypeptide of interest and N-terminally thereto a polypeptide exhibiting autoproteolytic function, said method comprising the steps of a) binding of the fusion polypeptide in a soluble, autoproteolytically inactive form by an affinity chromatography system, b) refolding of the fusion polypeptide, thereby activating the autoproteolytic function of the fusion polypeptide and causing cleavage of the heterologous polypeptide of interest, and c) subsequently eluting the heterologous polypeptide of interest, wherein said steps are conducted on one affinity chromatography system.
Type:
Application
Filed:
April 25, 2006
Publication date:
August 13, 2009
Applicants:
BOEHRINGER INGELHEIM AUSTRIA GMBH, SANDOZ AG
Inventors:
Alois Jungbauer, Rainer Hahn, Anne Tscheliessnig, Waltraud Kaar
Abstract: In a method for reconstituting a recombinant protein from a denatured state to its active form, a feed solution containing the recombinant protein in its denatured and/or in biologically inactive intermediate forms is subjected to a chromatographic separation process, in which the protein is reconstituted under conditions that promote refolding of the protein and the intermediate forms are separated from the refolded protein. The denatured form and/or the inactive intermediate forms of the protein are separated from the refolded protein in a continuous or quasi-continuous manner and optionally recycled to the feed solution.
Type:
Grant
Filed:
October 2, 2002
Date of Patent:
June 13, 2006
Assignee:
Boehringer Ingelheim Austria GmbH
Inventors:
Roman Necina, Robert Schlegl, Alois Jungbauer, Christine Machold
Abstract: A host-vector system that uses the RNA-based copy number control mechanism of ColE1-type plasmids for regulating the expression of a marker gene allows for antibiotic-free selection of plasmids and is useful for production of plasmid DNA and recombinant proteins.
Abstract: A process for producing plasmid DNA E. coli cells comprises a pre-culture and fed-batch process. The culture media of the batch phase and the culture medium added during the feeding phase are chemically defined. The culture medium of the feeding phase contains a growth-limiting substrate and is added, for at least a fraction of the feeding phase, at a feeding rate that follows a pre-defined exponential function, thereby controlling the specific growth rate at a pre-defined value. The process results in high yield and homogeneity of plasmid DNA.
Type:
Application
Filed:
April 8, 2005
Publication date:
October 20, 2005
Applicant:
Boehringer Ingelheim AUSTRIA GmbH
Inventors:
Hans Huber, Gerhard Weigl, Wolfgang Buchinger
Abstract: A method for producing plasmid DNA on a manufacturing scale uses Escherichia coli K-12 strain JM108. The process results in high yield and homogeneity of plasmid DNA.
Type:
Application
Filed:
April 8, 2005
Publication date:
October 20, 2005
Applicant:
Boehringer Ingelheim AUSTRIA GmbH
Inventors:
Hans Huber, Claudia Pacher, Roman Necina, Franz Kollmann, Christoph Reinisch
Abstract: A scalable process and device for producing a biomolecule, in particular pharmaceutical grade plasmid DNA. The process includes the steps of alkaline lysis and a neutralization. For separating the lysate and the precipitate, the mixture is allowed to gently flow downward through a clarification reactor that is partially filled, in its lower part, with retention material like glass beads, whereby the precipitate is retained on top of and within the retention. In a preferred embodiment of the lysis step, cell suspension and alkaline lysis solution flow through a lysis reactor that is filled with particulate material like glass beads. The process can be run continuosly and fully automated.
Type:
Application
Filed:
March 23, 2004
Publication date:
February 3, 2005
Applicant:
Boehringer Ingelheim Austria GmbH
Inventors:
Jochen Urthaler, Roman Necina, Christine Ascher, Helga Woehrer