Methods and compositions for the detection of bacterial endotoxins
First Claim
1. A method of producing an improved amebocyte lysate preparation having reduced Factor G activity, the method comprising the steps of:
- (a) admixing crude amebocyte lysate with a surfactant in an amount sufficient to produce a solution containing a precipitate; and
(b) separating said precipitate from said solution thereby to produce an amebocyte lysate preparation which is less reactive with (1→
3)-β
-D glucan than said crude amebocyte lysate.
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Abstract
The invention provides methods and compositions for the detection and/or quantification of bacterial endotoxins. In particular, provided herein is an inexpensive and reproducible method for producing an improved amebocyte lysate preparation having reduced Factor G activity. Provided also is an endotoxin-specific amebocyte lysate preparation produced by such a method. In addition, the invention provides methods and compositions for enhancing the sensitivity to endotoxins of amebocyte lysate preparations having reducing Factor G activity. In particular, the sensitivity of such amebocyte lysate preparations to endotoxins can be enhanced by the addition of exogenous (1→3) β-D-glucan.
16 Citations
33 Claims
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1. A method of producing an improved amebocyte lysate preparation having reduced Factor G activity, the method comprising the steps of:
-
(a) admixing crude amebocyte lysate with a surfactant in an amount sufficient to produce a solution containing a precipitate; and
(b) separating said precipitate from said solution thereby to produce an amebocyte lysate preparation which is less reactive with (1→
3)-β
-D glucan than said crude amebocyte lysate. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 26, 31)
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11. A method of producing an improved amebocyte lysate preparation having reduced Factor G activity, the method comprising the steps of:
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(a) admixing crude amebocyte lysate with a zwitterionic surfactant in an amount sufficient to produce a solution containing a precipitate; and
(b) removing said precipitate from said solution thereby to produce an amebocyte lysate preparation which is less reactive with (1→
3)-β
-D glucan than said crude amebocyte lysate. - View Dependent Claims (12, 13, 14, 15, 16, 17, 18, 19, 23, 24, 25, 27, 32)
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20. A method of producing an improved amebocyte lysate having reduced Factor G activity, the method comprising the steps of:
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(a) admixing crude amebocyte lysate with a n-tetradecyl-N, N-dimethyl-3-ammonio-1-propanesulfonate surfactant in an amount sufficient to produce a solution containing a precipitate;
(b) separating said precipitate from said solution; and
(c) removing said surfactant from said solution thereby to produce an amebocyte lysate preparation that is reactive with bacterial endotoxin and is less reactive with (1→
3)-β
-D glucan than said crude amebocyte lysate. - View Dependent Claims (21, 22)
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28. A composition comprising an amebocyte lysate preparation having reduced Factor G activity and exogenously added (1→
- 3)-β
-D glucan in an amount sufficient to enhance the sensitivity of said amebocyte lysate preparation to endotoxin relative to said amebocyte lysate preparation without said exogenously added (1→
3)-β
-D glucan. - View Dependent Claims (29, 30, 33)
- 3)-β
Specification