Endotoxin-binding ligands and their use

• US 8,765,393 B2
• Filed: 05/22/2003
• Issued: 07/01/2014
• Est. Priority Date: 05/22/2002
• Status: Active Grant

• Current Assignee: Astrea Uk Services Limited
• Original Assignee: Prometic Biosciences Ltd. (Thomvest Asset Management, Inc.)
• Inventors: Burton, Steven James, Podgorski, Tadeusz, Eapen, Saji S.
• Primary Examiner(s): Hines, Ja'na
• Application Number:
  US10/514,455
• Publication Number:
  US 20050238641A1
• Time in Patent Office: 4,058 Days
• Field of Search: None
• US Class Current: 435/7.32
• CPC Class Codes: A61L 2/0011 using physical methods
• CPC Class Codes: A61L 2/0017 Filtration
• CPC Class Codes: A61M 1/3486 Biological, chemical treatment, e.g. chemical precipitation; treatment by absorbents
• CPC Class Codes: A61M 1/3679 by absorption A61M1/3675 takes precedence
• CPC Class Codes: A61M 2202/20 Pathogenic agents
• CPC Class Codes: B01D 15/3804 Affinity chromatography
• CPC Class Codes: B01D 67/0093 Chemical modification
• CPC Class Codes: B01D 67/00931 by introduction of specific groups after membrane formation, e.g. by grafting
• CPC Class Codes: B01J 20/262 obtained otherwise than by reactions only involving carbon to carbon unsaturated bonds, e.g. obtained by polycondensation macromolecular compounds obtained otherwise than by reactions only involving unsaturated carbon-to-carbon bonds per se C08G
• CPC Class Codes: B01J 20/28016 Particle form
• CPC Class Codes: B01J 20/28033 Membrane, sheet, cloth, pad, lamellar or mat
• CPC Class Codes: B01J 20/3212 consisting of a polymer obtained by reactions otherwise than involving only carbon to carbon unsaturated bonds
• CPC Class Codes: B01J 20/3217 Resulting in a chemical bond between the coating or impregnating layer and the carrier, support or substrate, e.g. a covalent bond
• CPC Class Codes: B01J 20/3219 involving a particular spacer or linking group, e.g. for attaching an active group
• CPC Class Codes: B01J 20/3251 comprising at least two different types of heteroatoms selected from nitrogen, oxygen or sulphur
• CPC Class Codes: B01J 20/3255 comprising a cyclic structure containing at least one of the heteroatoms nitrogen, oxygen or sulfur, e.g. heterocyclic or heteroaromatic structures
• CPC Class Codes: B01J 2220/56 Use in the form of a bed
• CPC Class Codes: B01J 2220/58 Use in a single column
• CPC Class Codes: C02F 1/28 by sorption using ion-exchange C02F1/42; sorbent compositions B01J
• CPC Class Codes: C02F 2101/30 Organic compounds
• CPC Class Codes: C02F 2303/04 : Disinfection

Abstract

An affinity ligand-matrix conjugate of the structure Z-Spacer-[NX-A]_m-NY-A-NK₂ is useful for the isolation, separation, purification, characterization, identification or quantification of endotoxins in an aqueous system, wherein m is an integer of at least one; each A independently represents an optionally substituted linear, branched or cyclic saturated hydrocarbon chain containing 1 to 6 carbon atoms; each X independently represents hydrogen or alkyl; Y is X or A-NX₂; and Z is a support matrix attached to the ligand through an optional spacer arm (Spacer).

Claims

1. A method of selective removal of endotoxins from an aqueous system comprising one or more antibiotics and/or proteins wherein said method comprises the use of an affinity ligand-matrix conjugate of the structure

Z-Spacer-Ligand

wherein the ligand is tris(2-aminoethyl)amine;

wherein the ligand is attached to a support matrix Z through the spacer,

wherein said endotoxins are lipopolysaccharides (LPS) from gram-negative bacteria,

and wherein the method comprises;

contacting the affinity ligand-matrix conjugate with the aqueous system under conditions such that endotoxins in the aqueous system bind directly to the ligand, and

recovering the aqueous system that has been contacted with the affinity ligand matrix conjugate and from which endotoxins have been removed;

wherein essentially all of the protein and/or antibiotic remains in the aqueous system.

2. The method according to claim 1, wherein Z is selected from porous agarose beads, perfluorocarbon particles and functionalized membranes.

3. The method according to claim 1, wherein the aqueous system has a pH in the range of 3 to 10.

4. The method according to claim 1, wherein the aqueous system is contacted with a suspension or packed bed of the conjugate.

5. The method according to claim 1, wherein the aqueous system comprises whole blood or plasma and wherein the method comprises passing the aqueous system through an extracorporeal device comprising said affinity ligand-matrix conjugate under conditions such that endotoxins in the aqueous system bind directly to the ligand and are thereby removed from the aqueous system.

6. The method according to claim 1, wherein the ligand is attached to a soluble polymeric or non-polymeric carrier for in vivo capture of endotoxin.

7. The method according to claim 1, comprising providing an aqueous system comprising one or more antibiotics, wherein the recovered aqueous system comprises said one or more antibiotics.

8. The method according to claim 1, wherein Z is a porous agarose bead.

9. The method according to claim 1, wherein Z is a perfluorocarbon particle.

10. The method according to claim 1, wherein Z is a functionalized membrane.

11. The method according to claim 1, wherein at least 99% of the endotoxin is removed from the aqueous system.