Hybrid amebocyte lysate and uses thereof
AssigneeCharles River Laboratories, Inc.
InventorsNorman R. Wainwright, Masakazu Tsuchiya, John Dubczak, Foster T. Jordan
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Abstract
The invention relates generally to hybrid amebocyte lysate compositions (including both native and recombinant components) and their use in detecting and/or quantifying endotoxin in a sample.
Claims30
1-36. (canceled)
A method of preparing an amebocyte lysate composition comprising:
- (a) providing a recombinant factor C, a recombinant factor B, and a recombinant pro-clotting enzyme;
- and
- (b) combining the recombinant factor C, the recombinant factor B, and the recombinant pro-clotting enzyme to formulate the amebocyte lysate composition;
- wherein at least one of the recombinant factor B, recombinant factor C, or recombinant pro-clotting enzyme is made by expression in a host cell that has been genetically modified to prevent the addition of sialic acid to the recombinant factor B, the recombinant factor C, and/or the recombinant pro-clotting enzyme.
A composition comprising a recombinant factor C, a recombinant factor B, and a recombinant pro-clotting enzyme, wherein at least one of the recombinant factor B, the recombinant factor C, or the recombinant pro-clotting enzyme lacks sialic acid.
A method of producing a recombinant factor C protein comprising:
- (a) culturing a host cell transfected with a vector comprising a nucleic acid sequence encoding a factor C protein to obtain a cell culture;
- and
- (b) purifying the factor C protein from the cell culture;
- wherein the host cell is a host cell that has been genetically modified to prevent the addition of sialic acid to the factor C protein.
A host cell comprising a nucleic acid encoding a factor C protein, wherein the factor C protein comprises an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO:2, wherein the host cell has been genetically modified to prevent addition of sialic acid to the factor C protein.
Dependencies are inferred from the claim text.