A point-of-use kit is designed for optically detecting and quantifying bacterial endotoxin by employing specific formulations of Limulus amebocyte lysate (LAL), each formulation designed to optimize results with different sample classes. Kits are pre-certified for use with a variety of environmental, industrial, and clinical samples, each sample category having a unique kit design and containing a unique lysate reagent formulation. Pre-certification transfers time and reagent consuming tasks, such as assessment of sample compatibility and sample effect on reagent sensitivity to endotoxin from the user to the kit producer. A fixed dilution/sample treatment is employed, eliminating the need for a comparative water standard and for a sample positive control. The kit has LAL reagent prepackaged in dry polyethylene capped tubes, which retains reagent shelf life and optical clarity for accurate and reproducible results using a portable spectrophotometer/optical reader.

A process of using a fish plasma component in a nutrient medium for cell culture includes obtaining a fish that is a progeny of domesticated broodstock that are reared under consistent and reproducible conditions. Blood is obtained from the fish, and plasma is separated from the blood. One or more specific components of the plasma are then extracted, and cells are cultured in a nutrient medium using the one or more extracted plasma components, and none of any remainder of the plasma. The plasma and/or the plasma components is/are tested for presence and/or level of endotoxin. Extracting the one or more specific components of the plasma, and/or culturing the cells is only performed if the testing indicates an endotoxin level below a predetermined threshold. The cells cultured using the extracted one or more plasma components are other than fish cells.

A process of using a fish plasma component in a nutrient medium for cell culture includes obtaining a fish that is a progeny of domesticated broodstock that are reared under consistent and reproducible conditions. Blood is obtained from the fish, and plasma is separated from the blood. One or more specific components of the plasma are then extracted, and cells are cultured in a nutrient medium using the one or more extracted plasma components, and none of any remainder of the plasma. The plasma and/or the plasma components is/are tested for presence and/or level of endotoxin. Extracting the one or more specific components of the plasma, and/or culturing the cells is only performed if the testing indicates an endotoxin level below a predetermined threshold. The cells cultured using the extracted one or more plasma components are other than fish cells.

The present invention is directed to methods, compositions and devices useful for the detection and/or quantification of a microbial contaminant, including an endotoxin. In embodiments, cartridges are provided that suitably include dried compositions that are useful in absorbance-based assays and in combination with portable readers/devices.

The present invention is directed to methods, compositions and devices useful for the detection and/or quantification of a microbial contaminant, including an endotoxin. In embodiments, cartridges are provided that suitably include dried compositions that are useful in absorbance-based assays and in combination with portable readers/devices.

Provided is a technology related to a horseshoe crab factor B variant, and also provided is means for performing endotoxin measurement with high sensitivity. A polypeptide having an amino acid sequence in which the amino acid residue at the 193-position in an amino acid sequence of a polypeptide of horseshoe crab factor B is substituted with a cysteine (Cys) residue, is produced. Endotoxin measurement can be carried out with high sensitivity by configuring a Limulus reagent by combining this polypeptide with horseshoe crab factor C.

The present invention provides a novel method for the recombinant production of Factor C protein from horseshoe crab using a parasitic protozoan expressing the Factor C protein. In particular, the present invention provides a parasitic protozoan host cell harboring a polynucleotide encoding horseshoe crab Factor C protein, and a method for producing Factor C protein comprising culturing said parasitic protozoan host cell under conditions such that the cells express the horseshoe crab Factor C protein. Furthermore, the present invention provides recombinant Factor C protein produced by the novel method and its use in the detection and/or removal of endotoxin.

The present invention provides a novel method for the recombinant production of Factor C protein from horseshoe crab using a parasitic protozoan expressing the Factor C protein. In particular, the present invention provides a parasitic protozoan host cell harboring a polynucleotide encoding horseshoe crab Factor C protein, and a method for producing Factor C protein comprising culturing said parasitic protozoan host cell under conditions such that the cells express the horseshoe crab Factor C protein. Furthermore, the present invention provides recombinant Factor C protein produced by the novel method and its use in the detection and/or removal of endotoxin.

The present invention is related to compositions comprising clarified limulus amebocyte lysate (LAL), wherein the LAL is substantially free of coagulogen and methods of making such compositions. The invention further relates 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 clarified 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 invention also relates to kits comprising clarified LAL substantially free of coagulogen, and methods of making such.

The present invention is related to compositions comprising clarified limulus amebocyte lysate (LAL), wherein the LAL is substantially free of coagulogen and methods of making such compositions. The invention further relates 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 clarified 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 invention also relates to kits comprising clarified LAL substantially free of coagulogen, and methods of making such.