Reagent for endotoxin-specific assay

• US 5,702,882 A
• Filed: 09/28/1994
• Issued: 12/30/1997
• Est. Priority Date: 09/30/1993
• Status: Expired due to Term

• Current Assignee: Seikagaku Kogyo Kabushiki Kaisha
• Original Assignee: Seikagaku Kogyo Kabushiki Kaisha
• Inventors: Oda, Toshio, Tanaka, Shigenori, Tamura, Hiroshi
• Primary Examiner(s): Hollinden, Gary E.
• Assistant Examiner(s): LEARY, LOUISE N
• Application Number:
  US08/313,899
• Time in Patent Office: 1,189 Days
• Field of Search: 435/4, 435/18, 435/13, 435/23, 435/184, 435/975, 436/63, 436/74, 436/69, 436/14
• US Class Current: 435/4
• CPC Class Codes: G01N 33/579 involving limulus lysate
• CPC Class Codes: Y10S 435/975 Kit

Abstract

Endotoxin (Et) can be specifically assayed by exclusively utilizing the factor C system reaction without being affected by factor G contained in a limulus amebocyte lysate reagent.

The present invention provides: (1) a reagent for ET-specific assay which comprises a limulus amebocyte lysate reagent and an alkylglucoside; (2) a method of specifically assaying Et in a specimen using a limulus amebocyte lysate reagent, wherein an alkylglucoside is added to the limulus amebocyte lysate reagent and/or the specimen; and (3) a factor G activation inhibitor composition which comprises an alkylglucoside as an active ingredient capable of inhibiting the activation of factor G in limulus amebocyte by (1→3)-β-D-glucan.

Claims

1. A reagent for endotoxin-specific assay which comprises a limulus amebocyte lysate reagent and an alkylglucoside.

2. The reagent for endotoxin-specific assay according to claim 1, wherein said alkylglucoside is contained in an amount effective for inhibiting activation of factor G existing in said limulus amebocyte lysate reagent.

3. The reagent for endotoxin-specific assay according to claim 1, wherein an alkyl group contained in the alkylglucoside has from 1 to 30 carbon atoms.

4. The reagent for endotoxin-specific assay according to claim 1, wherein, in said alkylglucoside, glucose or glucose of a reducing end of a glucoside moiety is bound to an alkyl group via a glycosidic linkage.

5. The reagent for endotoxin-specific assay according to claim 4, wherein said alkylglucoside is an alkyl-O-β

-D-glucoside or an alkyl-S-β

-D-glucoside.

6. The reagent for endotoxin-specific assay according to claim 4, wherein said alkylglucoside is selected from the group consisting of n-octyl-β

-D-glucoside, n-heptyl-β

-D-glucoside, n-dodecyl-β

-D-glucoside, n-octyl-β

-D-thioglucoside and n-heptyl-β

-D-thioglucoside.

7. The reagent for endotoxin-specific assay according to claim 1, wherein said alkylglucoside is soluble or dispersible in water.

8. A method of assaying for endotoxin in a specimen using a limulus amebocyte lysate reagent, comprising the steps of:

(a) adding an alkylglucoside to the limulus amebocyte lysate reagent and/or the specimen, and

(b) assaying for endotoxin in the specimen.

9. The method of specifically assaying endotoxin according to claim 8, wherein the alkylglucoside is added in an amount effective for inhibiting activation of factor G existing in the limulus amebocyte lysate reagent.

10. The method of specifically assaying endotoxin according to claim 8, wherein, in said alkylglucoside, glucose or glucose of a reducing end of a glucoside moiety is bound to an alkyl group via a glycosidic linkage.

11. A kit for endotoxin-specific assay which comprises a limulus amebocyte lysate reagent and a reagent containing an alkylglucoside.

12. The kit according to claim 11, wherein said reagent containing alkylglucoside further contains a buffer agent.

13. The kit according to claim 11, which further comprises a reagent containing a substrate for clotting enzyme.

14. The kit according to claim 13, wherein said substrate is a coagulogen or a synthetic peptide substrate.

15. The kit for endotoxin-specific assay according to claim 11, wherein, in said alkylglucoside, glucose or glucose of a reducing end of a glucoside moiety is bound to an alkyl group via a glycosidic linkage.

16. A factor G activation inhibitor composition which comprises an alkylglucoside as an active ingredient capable of inhibiting the activation of factor G existing in limulus amebocyte by (1→

3)-β

-D-glucan and, water or a buffer agent free from endotoxin.

17. The factor G activation inhibitor composition according to claim 16, wherein, in said alkylglucoside, glucose or glucose of a reducing end of a glucoside moiety is bound to an alkyl group via a glycosidic linkage.

18. A method for inhibiting activation of factor G existing in a limulus amebocyte lysate reagent comprising the step of adding an alkylglucoside to the limulus amebocyte lysate reagent.

19. The method for inhibiting activation of factor G according to claim 18, wherein, in said alkylglycoside, glucose or glucose of a reducing end of a glucoside moiety is bound to an alkyl group via a glycosidic linkage.