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.
Claims11
A method of making limulus amebocyte lysate (LAL) substantially free of coagulogen, wherein the LAL substantially free of coagulogen comprises less than 5% (wt/wt) of coagulogen relative to total protein in the LAL as measured by SDS-PAGE with protein stain, comprising:
- subjecting a composition comprising LAL, wherein the LAL comprises coagulogen, to tangential flow filtration (TFF) using a 20 kDa to 50 kDa TFF filter, wherein the TFF produces a retentate and a filtrate;
- and
- collecting the retentate, thereby obtaining the LAL substantially free of coagulogen.
Dependencies are inferred from the claim text.