Limulus lysate of improved sensitivity and preparing the same
AssigneeAssociates of Cape Cod, Inc.
InventorsJames D. Sullivan, Jr., Stanley W. Watson
Life of this patent
Estimating the term…
Abstract
Limulus lysate of improved sensitivity to endotoxin is prepared by withdrawing hemolymph from Limulus polyphemus (horseshoe crab), preparing a lysate by lysing the amoebocytes obtained from the hemolymph, treating the lysate with an amount of an organic solvent sufficient to precipitate a clotting inhibitor within the lysate and isolating the organic solvent treated lysate. In another embodiment, the sensitivity of the lysate is further increased by adding a salt of calcium, magnesium, manganese, thereto.
Claims10
A method for preparing Limulus lysate of enhanced sensitivity to endotoxin, which comprises:
- withdrawing hemolymph from horseshoe crabs (Limulus);
- preparing a lysate by lysing the amoebocytes obtained from said hemolymph;
- treating said lysate with an amount of an organic solvent selected from the group consisting of chloroform, iodoform, bromoform, methylbromide, methylchloride, methyliodide, ethylchloride, ethylbromide, ethyliodide, propylchloride, propylbromide, propyliodide, ethylene chloride, methylene chloride, chlorobenzene, bromobenzene, iodobenzene, dimethylether, diethylether, carbontetrachloride, trichloroethylene, toluene and hexane sufficient to denature a clotting inhibitor present in said lysate;
- and
- isolating said organic solvent treated lysate.
A method for preparing Limulus lysate of enhanced sensitivity to endotoxin, which comprises the steps of:
- withdrawing hemolymph from horseshoe crabs (Limulus);
- preparing a lysate by lysing the amoebocytes obtained from said hemolymph;
- treating said lysate with an amount of an organic solvent selected from the group consisting of chloroform, iodoform, bromoform, methylbromide, methylchloride, methyliodide, ethylchloride, ethylbromide, ethyliodide, propylchloride, propylbromide, propyliodide, ethylene chloride, methylene chloride, chlorobenzene, bromobenzene, iodobenzene, dimethylether, diethylether, carbontetrachloride, trichloroethylene, toluene and hexane sufficient to denature a clotting inhibitor present in said lysate;
- isolating said organic solvent treated lysate;
- and
- adding a sensitivity increasing amount of a chloride salt of manganese, calcium or magnesium to said extracted lysate.
A lysate sensitive to endotoxins derived from horseshoe crabs (Limulus) by a process comprising:
- withdrawing hemolymph from horseshoe crabs (Limulus);
- preparing a lysate by lysing the amoebocytes obtained from said hemolymph;
- treating said lysate with an amount of an organic solvent selected from the group consisting of chloroform, iodoform, bromoform, methylbromide, methylchloride, methyliodide, ethylchloride, ethylbromide, ethyliodide, propylchloride, propylbromide, propyliodide, ethylenechloride, methylenechloride, chlorobenzene, bromobenzene, iodobenzene, dimethylether, diethylether, carbontetrachloride, trichloroethylene, toluene and hexane sufficient to denature a clotting inhibitor present in said lysate;
- and
- isolating said organic solvent treated lysate.
Dependencies are inferred from the claim text.