Process for inhibiting activity of endotoxin

• US 5,750,500 A
• Filed: 05/31/1995
• Issued: 05/12/1998
• Est. Priority Date: 01/21/1993
• Status: Expired due to Fees

• Current Assignee: WAKO Pure Chemical Industries Limited (Fujifilm Holdings Corporation)
• Original Assignee: WAKO Pure Chemical Industries Limited (Fujifilm Holdings Corporation)
• Inventors: Harada, Kazuaki, Tsuchiya, Masakazu
• Primary Examiner(s): Russel, Jeffrey E.
• Application Number:
  US08/455,563
• Time in Patent Office: 1,077 Days
• Field of Search: 530/317, 530/319, 530/326, 514/2, 514/9, 514/11, 514/21, 514/13, 435/7.21, 435/7.32, 435/18, 435/23, 435/34
• US Class Current: 514/2.4
• CPC Class Codes: G01N 33/56911 Bacteria
• CPC Class Codes: G01N 33/579 involving limulus lysate

Abstract

By contacting a peptide derivative or a protein having a property of binding to endotoxin (ET) to inhibit the activity of ET, and at least one surfactant with a sample containing ET, the activity of ET can be inhibited, and even when an ET analogous substance having ET activity is present in the thus treated sample, it can be measured without influence of ET.

Claims

1. A pretreating aqueous solution for inhibiting the activity of endotoxin consisting of water, an endotoxin-inhibiting peptide selected from the group consisting of polymyxins A, B, C, D, E, K, M and P, salts of those polymyxins, tachyplesin, polyphemusin and anti-LPS factor, and at least one surfactant, said solution being unable to react with a horseshoe crab hemocyte lysate, and serving not to inhibit or to enhance the reaction of horseshoe crab hemocyte lysate with an endotoxin analogous substance.

2. A pretreating solution according to claim 1, wherein the concentration of the surfactant is 0.01 to 10.0 w/v % and the concentration of the endotoxin-inhibiting peptide is 0.00002 to 0.2 w/v %.

3. A pretreating solution according to claim 1, wherein the surfactant is a nonionic surfactant or an amphoteric surfactant.

4. A pretreating solution according to claim 1, wherein the salt of polymyxin is polymyxin B sulfate.

5. An aqueous diluting solution for an endotoxin-containing sample, said solution serving to inhibit selectively the activity of the endotoxin in the sample, but not to inhibit the activity of an endotoxin analogous substance in the sample, to make only the latter responsive to horseshoe crab hemocyte lysate, said solution consisting of water, an endotoxin-inhibiting peptide selected from the group consisting of polymyxins A, B, C, D, E, K, M and P, salts of those polymyxins, tachyplesin, polyphemusin and anti-LPS factor, and at least one surfactant, said solution being unreactive with the horseshoe crab hemocyte lysate and not serving to inhibit or to enhance the reaction of the horseshoe crab hemocyte lysate with an endotoxin analogous substance, whereby after dilution with said solution, but before the addition of the horseshoe crab hemocyte lysate, the endotoxin activity in the endotoxin-containing sample is completely inhibited.

6. A diluting solution according to claim 5, wherein the concentration of the surfactant is 0.01 to 10.0 w/v % and the concentration of the endotoxin-inhibiting peptide is 0.00002 to 0.2 w/v %.

7. A diluting solution according to claim 5, wherein the surfactant is a nonionic surfactant or an amphoteric surfactant.

8. A diluting solution according to claim 5, wherein the salt of polymyxin is polymyxin B sulfate.