Congener independent detection of microcystin and nodularin congeners

• US 6,967,240 B1
• Filed: 09/06/2000
• Issued: 11/22/2005
• Est. Priority Date: 09/06/1999
• Status: Expired due to Term

• Current Assignee: Daniel R. Dietrich, New Zealand Pastoral Agriculture Research Institute Limited, Prof. Dr. Daniel R. Dietrich, Regents of the University of California (University of California)
• Original Assignee: Daniel R. Dietrich, New Zealand Pastoral Agriculture Research Institute Limited, Regents of the University of California (University of California)
• Inventors: Aggen, James B., Fischer, Werner, Garthwaite, Ian, Towers, Neale R., Miles, Christopher O., Dietrich, Daniel R., Chamberlin, A. Richard, Ross, Kathryn M.
• Primary Examiner(s): CEPERLEY, MARY
• Application Number:
  US10/070,302
• Time in Patent Office: 1,903 Days
• Field of Search: 436/531, 436/815, 436/161, 435/7.93, 435/7.95, 435/188, 530/389.8, 530/388.9, 530/403, 530/405, 530/406, 530/389.5, 530/388.4, 530/391.1
• US Class Current: 530/389.5
• CPC Class Codes: A61K 39/00 Medicinal preparations containing antigens or antibodies materials for immunoassay G01N33/53
• CPC Class Codes: C07K 14/195 from bacteria
• CPC Class Codes: C07K 16/12 against material from bacteria
• CPC Class Codes: C07K 7/56 the cyclisation not occurring through 2,4-diamino-butanoic acid

Abstract

The present invention relates to a protienaceous compound or functionally active derivative or part thereof having a binding site for a group represented by formula (I) which is part of a group of toxins derived from various cyanobacteria, to a method for its production, to diagnostic kits and to an affinty matrix (e.g. for use in immunoaffinity columns, online detection and purifications devices) containing the proteinaceous compound as well as to methods for substantially decreasing the amount of a compound containing the group represented by formula (I) in fluids or for concentrating compounds, e.g. toxins, containing the group represented by formula (I) from fluids such as crude water samples, extracts of algae or other tissue samples, e.g. to determine toxin concentrations.

Claims

1. A compound comprising one or more polypeptides providing a binding site of a monoclonal, polyclonal or recombinant antibody or a functionally active derivative or part thereof, wherein said compound is prepared using the group of formula (I) as a hapten, and said compound is capable of specifically binding to a compound having a structure of formula (I) represented as

wherein group R¹ represents a halogen atom, -OSO₃, -OR^′

or -NR′

₂ and group R² represents hydrogen, (C₁–

C₄)alkyl, (C₁–

C₄)alkoxy, (C₁–

C₄)acyl, (C₁–

C₄)acylamino, (C₁–

C₄)carboxyaminoacyl, glutamidyl, or 2-aminopropionamidyl, and wherein

the groups R³ which may be the same or different are each independently selected from the group consisting of hydrogen and (C₁–

C₄)alkyl,

group R⁴ represents (C₁–

C₄) alkoxy,

the phenyl group may be substituted or unsubstituted, and further wherein the groups R^′

represent hydrogen, substituted or unsubstituted (C₁–

C₄)alkyl or (C₁–

C₄)acyl.

2. The compound according to claim 1, wherein the groups R³ each represent methyl and group R⁴ represents methoxy.

5. The compound according to claim 1 or claim 3 which is a polyclonal, monoclonal or recombinant antibody or a functionally active derivative or fragment thereof.

6. A method for preparation of the compound according to claim 1 or claim 3, said method comprising the steps of:

(a) providing a compound containing a group represented by formula (I) as defined in claim 1 or claim 3;

(b) coupling the compound of step (a) to an immunogenic carrier to form a conjugate;

(c) immunizing an animal with the conjugate obtained in step (b); and

(d) isolating the animal'"'"'s blood, blood serum and/or spleenocytes.

7. The method according to claim 6, wherein the immunogenic carrier is a polymeric substance.

8. The method according to claim 7, wherein the polymeric substance is selected from the group consisting of polyethyleneglycol, polypeptides, proteins, polysaccharides and plastic supports.

9. The method according to claim 8, wherein the substance is a protein and said protein is selected from bovine serum albumin, ovalbumin, cationised bovine serum albumin or horseradish peroxidase.

10. A diagnostic kit containing the compound according to claim 1 or claim 3.

11. An affinity matrix containing the compound according to claim 1 or claim 3 coupled to a polymeric resin.

12. A method for detecting a compound containing a group represented by formula (I) as defined in claim 1 or claim 3, said method comprising the steps of:

(a) providing a compound according to claim 1 or claim 3;

(b) mixing a second compound suspected of containing a group represented by formula (I) as defined in claim 1 or claim 3 to form a mixture; and

(c) performing an assay that detects binding of the compound according to claim 1 or claim to the second compound.

13. A method for concentrating a compound containing a group represented by formula (I) as defined in claim 1 or claim 3 from a fluid or for substantially decreasing the amount of a compound containing the group represented by formula (I) in a fluid comprising the steps of:

(a) preparing the compound according to claim 1 or claim 3,

(b) coupling the compound obtained in step (a) to a polymeric matrix, and

(c) contacting the fluid with the polymeric matrix obtained in step (b).

14. The method according to claim 13, wherein the fluid is hemodialysis water, drinking water or water derived from rivers, lakes or oceans.

3. A compound comprising one or more polypeptides providing a binding site of a monoclonal, polyclonal or recombinant antibody or a functionally active derivative or part thereof, wherein said compound is prepared using the group of formula (I) as a hapten, and said compound is capable of specifically binding to a compound having a structure of formula (I) represented as

wherein group R¹ represents acylamino and group R² represents (C₁–

C₄)acyl;

or group R¹ represents glycyl or D-alanyl and group R² represents acetyl;

or group R¹ represents -NH₂ and group R² represents glutamidyl or 2-aminopropionamidyl, and wherein

the groups R³ which may be the same or different are each independently selected from the group consisting of hydrogen and (C₁–

C₄) alkyl,

group R⁴ represents (C₁-C₄)alkoxy,

the phenyl group may be substituted or unsubstituted, and further wherein the groups R¹ represent hydrogen, substituted or unsubstituted (C₁–

C₄)alkyl or (C₁–

C₄)acyl.

4. The compound according to claim 3, wherein the groups R³ each represent methyl and group R⁴ represents methoxy.