Method of Detecting an Endotoxin Using Limulus Amebocyte Lysate Substantially Free of Coagulogen
AssigneeLonza Walkersville, Inc.
InventorsCandice Stumbaugh, David S. HERBST, Kenneth E. NICHOLS, JR.
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Abstract
The present invention is related to a method of detecting an endotoxin in a sample using a chromogenic assay, the method comprising: (a) contacting the sample with a reagent comprising limulus amebocyte lysate (LAL) and a chromogenic substrate; and (b) measuring a chromogenic effect resulting from a change in the chromogenic substrate in the presence of endotoxin in the sample; wherein the LAL is substantially free of coagulogen. The method also relates to compositions and kits comprising LAL substantially free of coagulogen, and methods of making such.
Claims39
A method of detecting an endotoxin in a sample using a chromogenic assay, the method comprising:
- a. contacting the sample with a reagent comprising limulus amebocyte lysate (LAL) and a chromogenic substrate;
- b. measuring a chromogenic effect resulting from a change in the chromogenic substrate in the presence of endotoxin in the sample;
- wherein the LAL is substantially free of coagulogen.
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A method of detecting an endotoxin in a biological sample using a chromogenic assay, the method comprising:
- a. contacting the biological sample with an aqueous reagent comprising limulus amebocyte lysate (LAL) and Ac-Ile-Glu-Ala-Arg-pNA;
- b. measuring the change in absorbance at 405 nm resulting from the enzymatic cleavage of pNA from Ac-Ile-Glu-Ala-Arg-pNA in the presence of endotoxin in the sample;
- wherein the LAL is substantially free of coagulogen.
A composition comprising limulus amebocyte lysate (LAL) and a chromogenic substrate, wherein the LAL is substantially free of coagulogen.
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A method of making limulus amebocyte lysate (LAL) substantially free of coagulogen, the method comprising:
- a. providing a composition comprising LAL, wherein the LAL comprises coagulogen;
- b. adjusting the buffer;
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- c. subjecting the composition of (b) to tangential flow filtration using a 20 kDa to 50 kDa filter, thereby isolating LAL substantially free of coagulogen.
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Dependencies are inferred from the claim text.