Compositions and methods for producing high secreted yields of recombinant proteins
AssigneeBolt Threads, Inc.
InventorsJoshua Kittleson, Thomas Stevens, Rena Hill, Carlos Gustavo Pesce
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Abstract
The present disclosure relates to methods for producing recombinant proteins, as well as compositions used in and produced by such methods. Specifically, the present disclosure relates to methods for producing high secreted yields of recombinant proteins, and the compositions provided herein include recombinant host cells that comprise polynucleotide sequences encoding proteins operably linked to at least 2 distinct secretion signals.
Claims19
-38. (canceled)
A recombinant host cell comprising a first polynucleotide sequence and a second polynucleotide sequence encoding a structurally identical recombinant protein fused to distinct secretion signals,
- wherein said recombinant protein encoded by said first polynucleotide sequence is operably linked to a first secretion signal comprising a pre-region of an α-mating factor signal sequence of S. cerevisiae and a pro-region of an α-mating factor signal sequence of S. cerevisiae, wherein said first secretion signal comprises SEQ ID NO:
- 8 or a functional secretion signal variant thereof at least 90% identical to SEQ ID NO:
- 8, and
- wherein said recombinant protein encoded by said second polynucleotide sequence is operably linked to a second secretion signal comprising a pre-region of a DSE4 signal sequence of P. pastoris and a pro-region of an α-mating factor signal sequence of S. cerevisiae, wherein said second secretion signal comprises SEQ ID NO:
- 10 or a functional secretion signal variant thereof at least 90% identical to SEQ ID NO:
- 10.
A method for producing a recombinant protein, comprising the steps of:
- a) culturing a recombinant host cell of claim 1 in a culture medium to obtain a fermentation comprising said recombinant host cell;
- and
- b) extracting said recombinant protein from the culture medium.
A fermentation comprising the recombinant host cell of claim 1 and a culture medium.
Dependencies are inferred from the claim text.