Method for recombinant production of horseshoe crab Factor C protein in protozoa
AssigneeBioméRieux Deutschland GmbH
InventorsBernd Buchberger, Holger Grallert, Sonja Molinaro
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Abstract
The present invention provides a novel method for the recombinant production of Factor C protein from horseshoe crab using a parasitic protozoan expressing the Factor C protein. In particular, the present invention provides a parasitic protozoan host cell harbouring a polynucleotide encoding horseshoe crab Factor C protein, and a method for producing Factor C protein comprising culturing said parasitic protozoan host cell under conditions such that the cells express the horseshoe crab Factor C protein. Furthermore, the present invention provides recombinant Factor C protein produced by the novel method and its use in the detection and/or removal of endotoxin.
Claims10
A method for detecting endotoxin in a sample suspected of containing endotoxin, the method comprising
- a) obtaining a recombinant horseshoe crab Factor C prepared by culturing a parasitic protozoan comprising a polynucleotide encoding horseshoe crab Factor C protein, said protozoan expressing a two-chain zymogen form of horseshoe crab Factor C protein having a molecular weight of about 102 kDa as determined by SDS-PAGE under non-reducing conditions, said zymogen being rendered enzymatically active upon contacting lipopolysaccharide and recovering the recombinant Factor C;
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- b) contacting the sample to be assayed for the presence of endotoxin with the recombinant horseshoe crab Factor C by measuring serine protease activity of the recombinant Factor C.
Dependencies are inferred from the claim text.