Methods for producing thrombin
AssigneeZymoGenetics, Inc.
InventorsRichard D. Holly, Donald C. Foster
Life of this patent
Estimating the term…
Abstract
Methods are disclosed for producing thrombin. The protein is produced from host cells transformed or transfected with DNA construct(s) containing information necessary to direct the expression of thrombin precursors. The DNA constructs generally include the following operably linked elements: a transcriptional promoter, DNA sequence encoding a gla-domainless prothrombin, and a transcriptional terminator. Thrombin precursors produced from transformed or transfected host cells are activated either in vivo or in vitro.
Claims31
A DNA construct which directs the expression of a thrombin precursor and comprising the following operably linked elements:
- a transcriptional promoter, a DNA segment encoding a gla-domainless prothrombin and a transcriptional terminator.
A host cell into which has been introduced a DNA construct which directs the expression of a thrombin precursor, said construct comprising the following operably linked element:
- a transcriptional promoter, a DNA sequence encoding a gla-domainless prothrombin and a transcriptional terminator.
A method for producing thrombin precursor comprising the steps of:
- growing a host cell, into which has been introduced a DNA construct which directs the expression of a thrombin precursor and which DNA construct comprises an operably linked transcriptional promoter, a DNA segment encoding a gla-domainless prothrombin and a transcriptional terminator, under suitable conditions to allow the expression of the thrombin precursor encoded by the DNA segment;
- and
- isolating the thrombin precursor from the host cell.
A method for producing thrombin comprising the steps of:
- growing a host cell, into which has been introduced a DNA construct which directs the expression of a thrombin precursor and which comprises an operably linked transcriptional promoter, a DNA segment encoding a gla-domainless prothrombin and a transcriptional terminator, under suitable conditions to allow the expression of the thrombin precursor encoded by the DNA segment and its activation to thrombin in the host cell;
- and
- isolating the activated thrombin from the host cell.
A method for producing thrombin comprising the steps of:
- growing a host cell, into which has been introduced a DNA construct which directs the expression of a thrombin precursor and which comprises an operably linked transcriptional promoter, a first DNA segment encoding a signal sequence joined to a second DNA segment encoding a gla-domainless prothrombin and a transcriptional terminator, in a suitable medium and under conditions to allow the expression of the thrombin precursor encoded by said second DNA segment;
- activating the thrombin precursor to thrombin;
- and
- isolating the thrombin from the medium.
Dependencies are inferred from the claim text.