DETECTING COMPLEMENT ACTIVATION

• US 20120315266A1
• Filed: 05/01/2012
• Published: 12/13/2012
• Est. Priority Date: 11/02/2010
• Status: Active Grant

• Current Assignee: Kypha, Inc.
• Original Assignee: Kypha, Inc.
• Inventors: Olson, Paul, Moss, Don W., Staten, Nicholas
• Granted Patent: US 8,865,164 B2
• Time in Patent Office: Days
• US Class Current: 424/130.1
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 29/00 Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
• CPC Class Codes: A61P 31/00 Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: G01N 2333/4716 Complement proteins, e.g. anaphylatoxin, C3a, C5a
• CPC Class Codes: G01N 2800/104 Lupus erythematosus [SLE]
• CPC Class Codes: G01N 2800/125 Adult respiratory distress syndrome
• CPC Class Codes: G01N 2800/26 Infectious diseases, e.g. generalised sepsis
• CPC Class Codes: G01N 2800/52 Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
• CPC Class Codes: G01N 33/5306 Improving reaction conditions, e.g. reduction of non-specific binding, promotion of specific binding
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9

Abstract

Methods of detecting complement activation including steps of detecting in a sample from a subject a level of iC3b wherein the detecting involves specific interaction between the iC3b and a non-cross-reactive antibody thereto, comparing the detected level with a reference level, which reference level is within a range of about 10 ng/ml to about 5,000 ng/ml, wherein determination that the detected level is above the reference level indicates that the subject is suffering from or susceptible to undesirable and/or pathologic complement activation, and administering treatment to treat undesired complement activation if the detected level is above the reference level. Other methods of detecting complement activation with or without measuring iC3b are also provided.

Claims

1. A method comprising steps of

detecting in a sample from a subject a level of intact C3, wherein the detecting involves specific interaction between the intact C3 and a non-cross-reactive antibody thereto;

comparing the detected level with a reference level, which reference level is within a range of about 350 ug/ml to about 1,700 ug/ml;

wherein determination that the detected level is below the reference level indicates that the subject is suffering from or susceptible to undesirable and/or pathologic complement activation; and

administering treatment to treat undesired complement activation if the detected level is below the reference level.

2. The method of claim 1, wherein the non-cross-reactive antibody is characterized in that a 1 ug/ul solution of iC3b produces a signal equivalent to less than about 1 ng/ml of C3.

3. The method of claim 2, wherein the non-cross-reactive antibody is or comprises HM2075.

4. The method of claim 1, wherein the detecting and comparing steps may be carried out in about 30 minutes or less.

5. The method of claim 1, wherein the undesirable and/or pathologic complement activation is caused by a disorder selected from the group consisting of systemic lupus erythematosus, trauma, inflammatory stress, autoimmune disorders, intracranial hemorrhage, infection, transplant rejection, ocular disease, heart disease, ischemia/reperfusion injury, age-related macular degeneration, paroxysmal nocturnal hemoglobinuria, hereditary angiodema, renal disease, pregnancy-associated disorders, and neurological disorders.

6. The method of claim 1, wherein the sample is selected from the group consisting of whole blood, serum, plasma, urine, tears, saliva, wound exudate, bronchioalveolar lavage fluid, and cerebrospinal fluid.

7. The method of claim 1, wherein the step of detecting is performed under controlled conditions such that performance of the step does not substantially activate complement within the sample.

8. The method of claim 1, wherein the detecting and comparing steps are not carried out using an ELISA assay.

9. The method of claim 1, wherein the detecting and comparing steps are carried out using a lateral flow assay.

10. A method comprising steps of

detecting in a sample from a subject a level of iC3b wherein the detecting involves specific interaction between the iC3b and a non-cross-reactive antibody thereto;

comparing the detected level with a reference level, which reference level is within a range of about 10 ng/ml to about 5,000 ng/ml;

wherein determination that the detected level is above the reference level indicates that the subject is suffering from or susceptible to undesirable and/or pathologic complement activation; and

administering treatment to treat undesired complement activation if the detected level is above the reference level.

11. The method of claim 10, wherein the non-cross-reactive antibody is characterized in that a 1 ug/ul solution of C3 produces a signal equivalent to less than about 1 ng/ml of iC3b.

12. The method of claim 10, wherein the non-cross-reactive antibody is or comprises an antibody selected from the group consisting of A209, MCA2607, and HM2199.

13. The method of claim 10, wherein the detecting and comparing steps may be carried out in about 30 minutes or less.

14. The method of claim 10, wherein the undesirable and/or pathologic complement activation is caused by a disorder selected from the group consisting of systemic lupus erythematosus, trauma, inflammatory stress, autoimmune disorders, intracranial hemorrhage, infection, transplant rejection, ocular disease, heart disease, ischemia/reperfusion injury, age-related macular degeneration, paroxysmal nocturnal hemoglobinuria, hereditary angiodema, renal disease, pregnancy-associated disorders, and neurological disorders.

15. The method of claim 10, wherein the sample is selected from the group consisting of whole blood, serum, plasma, urine, tears, saliva, wound exudate, bronchioalveolar lavage fluid, and cerebrospinal fluid.

16. The method of claim 10, wherein the step of detecting is performed under controlled conditions such that performance of the step does not substantially activate complement within the sample.

17. The method of claim 10, wherein the detecting and comparing steps are not carried out using an ELISA assay

18. The method of claim 10, wherein the detecting and comparing steps are carried out using a lateral flow assay.

19. A method comprising the steps of

detecting in a sample a ratio of intact C3 level to iC3b level, wherein the detecting involves specific interaction between the intact C3, the iC3b, or both with a non-cross-reactive antibody thereto;

comparing the detected level with a reference ratio of about 0.001 wherein determination that the detected level is below the reference level indicates that the subject is suffering from or susceptible to undesirable and/or pathologic complement activation; and

administering treatment to treat undesired complement activation if the detected level is below the reference level.

20. The method of claim 19, wherein the detecting and comparing steps may be carried out in about 30 minutes or less.

21. The method of claim 19, wherein the non-cross-reactive antibody for detecting intact C3 is or comprises HM2075.

22. The method of claim 19, wherein the non-cross-reactive antibody for detecting iC3b is or comprises an antibody selected from the group consisting of A209, MCA2607, and HM2199.

23. The method of claim 19, wherein the undesirable and/or pathologic complement activation is caused by a disorder selected from the group consisting of systemic lupus erythematosus, trauma, inflammatory stress, autoimmune disorders, intracranial hemorrhage, infection, transplant rejection, ocular disease, heart disease, ischemia/reperfusion injury, age-related macular degeneration, paroxysmal nocturnal hemoglobinuria, hereditary angiodema, renal disease, pregnancy-associated disorders, and neurological disorders.

24. The method of claim 19, wherein the sample is selected from the group consisting of whole blood, serum, plasma, urine, tears, saliva, wound exudate, bronchioalveolar lavage fluid, and cerebrospinal fluid.

25. The method of any one of claims 19, wherein the step of detecting is performed under controlled conditions such that performance of the step does not substantially activate complement within the sample.

26. The method of claim 19, wherein the detecting and comparing steps are not carried out using an ELISA assay

27. The method of claim 19, wherein the detecting and comparing steps are carried out using a lateral flow assay.