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US 12,612,638 B2Granted
Fusion constructs to express biopharmaceutical polypeptides in cyanobacteria
AssigneeThe Regents of the University of California
InventorsAnastasios Melis, Nico Betterle, Diego Hidalgo Martinez, Andrew C. Saphire
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Abstract
This invention provides compositions and methods for providing high product yield of transgenes encoding biopharmaceutical polypeptides in cyanobacteria and microalgae.
Claims14
A cyanobacterial host cell comprising an expression unit comprising:
- (i) a nucleic acid sequence comprising a transgene that encodes a biopharmaceutical protein, wherein the transgene is fused to the 3′ end of a nucleic acid sequence that encodes a cyanobacteria β-subunit of phycocyanin (cpcB) polypeptide to produce a fusion polypeptide that comprises cpcB and the biopharmaceutical protein, and wherein the biopharmaceutical protein comprises:
- an interferon polypeptide having at least 95% identity to SEQ ID NO:
- 1,
- a tPA polypeptide having at least 95% identity to the region of SEQ ID NO:
- 2 that lacks the signal peptide or having at least 95% identity to SEQ ID NO:
- 3,
- a TTFC polypeptide having at least 95% identity to SEQ ID NO:
- 15,
- a Cholera Toxin Fragment B polypeptide having at least 95% identity to SEQ ID NO:
- 18;
- an insulin polypeptide having at least 95% identity to SEQ ID NO:
- 4, or
- a SARS-CoV2 polypeptide having at least 95% identity to SEQ ID NO:
- 16 or 17;
- (ii) a nucleic acid sequence encoding a cyanobacteria α-subunit of phycocyanin (cpcA) polypeptide;
- and
- (iii) a nucleic acid sequence encoding a cyanobacterial cpcC1, cpcC2 and cpcD polypeptide.
14 more elements
A nucleic acid encoding a fusion polypeptide that comprises cpcB and a biopharmaceutical protein, and wherein the biopharmaceutical protein comprises:
- an interferon polypeptide having at least 95% identity to SEQ ID NO:
- 1,
- a tPA polypeptide having at least 95% identity to the region of SEQ ID NO:
- 2 that lacks the signal peptide or having at least 95% identity to SEQ ID NO:
- 3,
- a TTFC polypeptide having at least 95% identity to SEQ ID NO:
- 15,
- a Cholera Toxin Fragment B polypeptide having at least 95% identity to SEQ ID NO:
- 18;
- an insulin polypeptide having at least 95% identity to SEQ ID NO:
- 4, or
- a SARS-CoV2 polypeptide having at least 95% identity to SEQ ID NO:
- 16 or 17.
10 more elements
Dependencies are inferred from the claim text.