Optimized expression cassette for expressing a polypeptide with high yield
AssigneeNovartis AG
InventorsZorica Dragic, Sabine Geisse, Rita Schmitz, Burkhard Wilms
Life of this patent
Estimating the term…
Abstract
The present invention is based on the finding that the combination of a specific 5′UTR polynucleotide sequence (see SEQ ID NO 1) and the hCD33 secretory leader sequence in an expression cassette for expressing a polypeptide of interest results in a surprisingly better expression level of the polypeptide of interest compared to prior art expression cassettes. Based on this finding, the present invention inter alia provides novel expression cassettes, expression vectors and methods for producing a polypeptide of interest with high yield.
Claims41
An expression cassette for stably expressing a polypeptide of interest comprising:
- a) a promoter;
- b) a 5′UTR polynucleotide which comprises a sequence selected from the group consisting of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, and SEQ ID NO:
- 5;
- c) a polynucleotide encoding a hCD33 secretory leader sequence;
- and
- d) a polynucleotide encoding a polypeptide of interest or an insertion site for inserting a polynucleotide encoding a polypeptide of interest,
- wherein the polypeptide of interest is expressed at a higher yield and/or with reduced incorrect signal peptide processing compared to expression of the polypeptide of interest from an identical control expression cassette except that the hCD33 secretory leader sequence has been replaced with an immunoglobulin secretory leader sequence.
An expression cassette for expressing a polypeptide of interest comprising:
- a) a promoter;
- b) a 5′UTR polynucleotide comprising the sequence of SEQ ID NO:
- 4 or SEQ ID NO:
- 5;
- c) a polynucleotide encoding a hCD33 secretory leader sequence;
- and
- d) a polynucleotide encoding a polypeptide of interest or an insertion site for inserting a polynucleotide encoding a polypeptide of interest.
An expression cassette for stably expressing a polypeptide of interest comprising:
- a) a promoter;
- b) a 5′UTR polynucleotide which consists of a sequence selected from the group consisting of SEQ ID NO:
- 1, SEQ ID NO:
- 2, SEQ ID NO:
- 3, SEQ ID NO:
- 4, and SEQ ID NO:
- 5;
- c) a polynucleotide encoding a hCD33 secretory leader sequence;
- and
- d) a polynucleotide encoding a polypeptide of interest or an insertion site for inserting a polynucleotide encoding a polypeptide of interest
- wherein the polypeptide of interest is expressed at a higher yield and/or with reduced incorrect signal peptide processing compared to expression of the polypeptide of interest from an identical control expression cassette except that the hCD33 secretory leader sequence has been replaced with an immunoglobulin secretory leader sequence.
Dependencies are inferred from the claim text.