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US 11,959,109 B2Granted
Recombinant Factor C and method for producing the same, and method for measuring endotoxin
AssigneeSeikagaku Corporation
InventorsHikaru Mizumura, Toshio Oda, Shun-ichiro Kawabata
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Abstract
A horseshoe crab Factor C protein having activity of Factor C, wherein the horseshoe crab is selected from Tachypleus tridentatus, Limulus polyphemus, and Carcinoscorpius rotundicauda, and wherein the horseshoe crab Factor C protein is produced through being recombinantly expressed from a Chinese Hamster Ovary (CHO) DG44 cell or HEK cell.
Claims10
A method for producing an endotoxin assay agent comprising a horseshoe crab Factor C, a horseshoe crab Factor B, and a horseshoe crab Pro-clotting enzyme, the method comprising:
- expressing the horseshoe crab Factor C in a Chinese Hamster Ovary cell or a Human Embryonic Kidney cell as a host cell, secreting the horseshoe crab Factor C into an extracellular culture supernatant, and recovering the horseshoe crab Factor C in the culture supernatant, wherein the recovered horseshoe Factor C has activity of Factor C, and wherein the horseshoe crab Factor C is a Tachypleus tridentatus Factor C or a Limulus polyphemus Factor C,
- expressing the horseshoe crab Factor B having activity of Factor B in a mammalian cell as a host cell,
- expressing the horseshoe crab Pro-clotting enzyme in a mammalian cell as a host cell, and
- producing an endotoxin assay agent comprising the horseshoe crab Factor C, the horseshoe crab Factor B, and the horseshoe crab Pro-clotting enzyme,
- wherein the host cell for expressing the horseshoe crab Factor C, the host cell for expressing the horseshoe crab Factor B, and the host cell for expressing the horseshoe crab Pro-clotting enzyme may be the same or different.
2 more elements
A method for measuring endotoxin in a test specimen, the method comprising
- expressing a horseshoe crab Factor C in a Chinese Hamster Ovary cell or a Human Embryonic Kidney cell as a host cell, secreting the horseshoe crab Factor C into an extracellular culture supernatant, and recovering the horseshoe crab Factor C in the culture supernatant, wherein the recovered horseshoe Factor C has activity of Factor C, and wherein the horseshoe crab Factor C is a Tachypleus tridentatus Factor C or a Limulus polyphemus Factor C,
- expressing a horseshoe crab Factor B having activity of Factor B in a mammalian cell as a host cell,
- expressing a horseshoe crab Pro-clotting enzyme in a mammalian cell as a host cell,
- producing an endotoxin assay agent comprising the horseshoe crab Factor C, the horseshoe crab Factor B, and the horseshoe crab Pro-clotting enzyme,
- mixing the endotoxin assay agent and a test specimen, and
- measuring progress of a cascade reaction,
- wherein the host cell for expressing the horseshoe crab Factor C, the host cell for expressing the horseshoe crab Factor B, and the host cell for expressing the horseshoe crab Pro-clotting enzyme may be the same or different.
4 more elements
Dependencies are inferred from the claim text.