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
To provide a method for producing a horseshoe crab recombinant Factor C. The horseshoe crab recombinant Factor C is produced through expression thereof by use of mammalian cells such as CHO DG44 and HEK293 as host cells.
Claims20
A horseshoe crab Factor C protein having activity of Factor C, which contains (α-2,3)-linked terminal sialic acid in a greater amount, as compared with a corresponding native horseshoe crab Factor C protein and/or a corresponding horseshoe crab Factor C protein recombinantly expressed using Sf9 as a host cell,
- wherein the horseshoe crab is selected from the group consisting of Tachypleus tridentatus, Limulus polyphemus, and Carcinoscoryius rotundicauda, and wherein the horseshoe crab Factor C protein is produced through being recombinantly expressed from a Chinese Hamster Ovary (CHO) DG44 cell.
A Factor C protein having activity of Factor C, wherein the Factor C protein contains (α-2,3)-linked terminal sialic acid in a greater amount, as compared with a corresponding native horseshoe crab Factor C protein;
- wherein the Factor C protein comprises an amino acid sequence selected from the group consisting of:
- (A) the amino acid sequence of SEQ ID NO:
- 2;
- (B) an amino acid sequence having 90% or higher sequence identity to the amino acid sequence of SEQ ID NO:2, and having activity of Factor C;
- (C) the amino acid sequence of SEQ ID NO:
- 4;
- and
- (D) an amino acid sequence having 90% or higher sequence identity to the amino acid sequence of SEQ ID NO:4, and having activity of Factor C,
- wherein the Factor C protein is produced through being recombinantly expressed from a CHO DG44 cell, and
- wherein the Factor C protein exhibits a residual activity of 10% or higher in the presence of 21 mM sodium citrate.
A horseshoe crab Factor C protein having activity of Factor C, wherein the horseshoe crab Factor C protein contains (α-2,3)-linked terminal sialic acid in a greater amount, as compared with a corresponding native horseshoe crab Factor C protein and/or a horseshoe crab Factor C protein recombinantly expressed using Sf9 as a host cell,
- wherein the horseshoe crab Factor C protein comprises an amino acid sequence selected from the group consisting of:
- (A) the amino acid sequence of SEQ ID NO:
- 2;
- and
- (B) the amino acid sequence of SEQ ID NO:
- 4,
- wherein the horseshoe crab Factor C protein is produced through being recombinantly expressed from a CHO DG44 cell, and
- wherein the horseshoe crab Factor C protein exhibits a residual activity of 10% or higher in the presence of 21 mM sodium citrate.
Dependencies are inferred from the claim text.