Reagent for endotoxin assay and method for endotoxin assay using the same

• US 5,840,510 A
• Filed: 06/30/1997
• Issued: 11/24/1998
• Est. Priority Date: 02/26/1993
• Status: Expired due to Term

• Current Assignee: Seikagaku Kogyo Kabushiki Kaisha
• Original Assignee: Seikagaku Kogyo Kabushiki Kaisha
• Inventors: Aita, Kazuhiro, Tanaka, Shigenori, Tamura, Hiroshi
• Primary Examiner(s): Leary, Louise N.
• Application Number:
  US08/885,176
• Time in Patent Office: 512 Days
• Field of Search: 435/23, 435/13, 435/4, 435/18, 435/19, 435/24, 435/29, 435/7.9
• US Class Current: 435/18
• CPC Class Codes: G01N 2333/811 Serine protease (E.C. 3.4.21) inhibitors
• CPC Class Codes: G01N 33/579 involving limulus lysate
• CPC Class Codes: Y10S 435/967 Standards, controls, materials, e.g. validation studies, buffer systems

Abstract

This invention provides (1) a reagent for endotoxin assay which comprises aprotinin and a limulus amebocyte lysate reagent, (2) a kit for endotoxin assay which comprises the limulus amebocyte lysate reagent and a reagent containing aprotinin, (3) a method for assaying endotoxin in a sample using the limulus amebocyte lysate reagent in which aprotinin is added to the lysate reagent and/or the sample, (4) a method for assaying endotoxin in a serine protease-containing sample using the limulus amebocyte lysate reagent in which the sample is allowed to contact with an aprotinin-immobilized insoluble carrier in advance of endotoxin assay, (5) a carrier for pretreating a serine protease-containing sample on which aprotinin is immobilized, (6) a method for inhibiting factor G activation in which aprotinin is added to the limulus amebocyte lysate reagent and (7) a factor G activation inhibitor which comprises aprotinin as an active ingredient.

Endotoxin assay can be effected based on the factor C system reaction, without influences of factor G contained in the limulus amebocyte lysate reagent and/or serine proteases contained in samples.

Claims

1. A reagent composition for specifically assaying for endotoxin in a sample comprising:

(A) aprotinin; and

(B) a horseshoe crab amebocyte lysate reagent, wherein said aprotinin is present in the reagent composition in an amount sufficient to completely inhibit activation of factor G present in the horseshoe crab amebocyte lysate reagent.

2. A kit for specifically assaying for endotoxin in a sample comprising:

(A) a horseshoe crab amebocyte lysate reagent; and

(B) a reagent containing aprotinin in an amount sufficient to completely inhibit activation of factor G present in the horseshoe crab amebocyte lysate reagent.

3. The kit as claimed in claim 2, wherein said reagent (B) comprises a buffer agent.

4. The kit as claimed in claim 2, wherein said lysate reagent is lyophilized, and said kit includes a solution for dissolving said lyophilized lysate reagent.

5. The kit as claimed in claim 2, wherein said reagent (B) is lyophilized, and said kit includes a solution for dissolving said lyophilized reagent (B).

6. The kit as claimed in claim 2, wherein said kit additionally comprises a lyophilized standard endotoxin, and a solution for dissolving said lyophilized standard endotoxin.