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Limulus polyphemus and other horseshoe crabs defend themselves against infection and physical damage with a clot-forming, zymogen-cascade. The inactive
components of the cascade are sequestered in intracellular granules of the circulating amebocytes . When the hemolymph comes in contact with the lipopolysaccharide (LPS) of Gram-negative bacterial cell walls or（1-3）-β-D-glucans of fungal cell walls , the amebocytes degranulate, releasing the proteins of the cascade.The LPS then activates Factor C, while the （1 -3）-β-D-glucan activates Factor G of the zymogen cascade resulting in clot formation from the cleavage of coagulogen. Amebocytes also contain an LPS-binding protein known as the Limulus anti-lipopolysaccharide factor (LALF) . The amino-acid sequence of L.
polyphemus LALF was the basis for making thesynthetic gene, the product of which is called endotoxin neutralizing protein (rENP). The natural product
(LALF) and rENP are sold commercially for neutralizing endotoxin either as puriﬁed proteins or covalently attached to solid-support devices.

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