A NOVEL GENERATION OF CLONED HORSESHOE CRAB RECOMBINATION FACTOR C FOR DETECTION AND REMOVAL OF ENDOTOXIN

摘要

The horseshoe crab, Carcinoscropius rotundicauda FactorC cDNA (CrFC21) has been cloned into a shuttle baculoviralvector. The recombinant baculoviral DNA was then transfectedinto the insect cells for expression of recombinant Factor C.Recombinant Factor C was found to be immunoreactive and iscapable of binding both free and bound/immobilized lipid A. It isenzymatically active when triggered by LPS. The rFC is probablyof the two-chain form, being cleaved into the heavy and lightchains after activation by Gram negative bacterial endotoxin. Aslow as 0.01 pg (0.001 ng/ml) of LPS was detectable by the rFC,thus, indicating its potentials as a novel generation of "limulusamoebocyte lysate".