Methods, compositions, and kits for detection of aspergillosis

Patent metadata

Field	Value
Publication number	US-9416395-B2
Application number	US-201414217004-A
Country	US
Kind code	B2
Filing date	Mar 17, 2014
Priority date	Mar 15, 2013
Publication date	Aug 16, 2016
Grant date	Aug 16, 2016

Abstract

Provided herein are methods for detecting an Aspergillus protease in a sample, diagnosing a subject with aspergillosis caused by an Aspergillus infection based on the presence of an Aspergillus protease in a sample, and methods of aspergillosis treatment that incorporate these diagnostic methods. In certain embodiments, the Aspergillus protease is Asp f2, and the Aspergillus infection is caused A. fumigatus, A. flavus, A. versicolor, A. niger , or A. terreus . Also provided herein are antibodies and kits for use in these methods, including novel antibodies specific for Asp f2.

First claim

What is claimed is:

1. A method for detecting Aspergillus Asp f2 in a biological sample comprising contacting the sample with an Asp f2 protease substrate that can be cleaved by Asp f2,
wherein cleavage of the protease substrate indicates the presence of Asp f2 in the sample, and
wherein the sample is enriched for Asp f2 prior to contact with the Asp f2 protease substrate by contacting the sample with an Asp f2-specific antibody or antigen binding fragment, the Asp f2-specific antibody or antigen binding fragment comprising one or more CDRs selected from the group consisting of SEQ ID NOs: 8, 10, 12, 14, 16 and 18.

2. A method for diagnosing aspergillosis in a subject comprising:
(a) obtaining a biological sample from the subject;
(b) detecting the presence of Aspergillus Asp f2 in the sample by contacting the sample with an Asp f2 protease substrate that can be cleaved by Asp f2, wherein cleavage of the protease substrate indicates the presence of Asp f2 in the sample; and
(c) diagnosing the subject with aspergillosis if Asp f2 is present in the sample,
wherein the sample is enriched for Asp f2 prior to contact with the Asp f2 protease substrate by contacting the sample with an Asp f2-specific antibody or antigen binding fragment, the Asp f2-specific antibody or antigen binding fragment comprising one or more CDRs selected from the group consisting of SEQ ID NOs: 8, 10, 12, 14, 16 and 18.

3. The method of claim 2 , further comprising (d) administering one or more therapeutic agents for the treatment of aspergillosis.

4. A method for the treatment of aspergillosis in a subject in need thereof comprising:
(a) obtaining a biological sample from the subject;
(b) detecting the presence of Aspergillus Asp f2 in the sample by contacting the sample with an Asp f2 protease substrate that can be cleaved by Asp f2, wherein cleavage of the protease substrate indicates the presence of Asp f2 in the sample; and
(c) administering one or more therapeutic agents for the treatment of aspergillosis if Asp f2 is detected in the sample, and
wherein the sample is enriched for Asp f2 prior to contact with the Asp f2 protease substrate by contacting the sample with an Asp f2-specific antibody or antigen binding fragment, the Asp f2-specific antibody or antigen binding fragment comprising one or more CDRs selected from the group consisting of SEQ ID NOs: 8, 10, 12, 14, 16 and 18.

5. The method of any one of claims 1 , 2 , and 4 , wherein the Asp f2-specific antibody is immobilized on an enrichment matrix.

6. The method of any one of claims 1 , 2 , and 4 , wherein the protease substrate is selected from the group consisting of a protein or peptide comprising the amino acid sequence of SEQ ID NO: 5, an elastin protein or peptide, and a collagen protein or peptide.

7. The method of any one of claims 1 , 2 , and 4 , wherein the protease substrate is conjugated to one or more fluorophores.

8. The method of claim 1 or 2 , wherein the aspergillosis is the result of infection with an Aspergillus species selected from the group consisting of A. fumigatus, A. flavus, A. versicolor, A. niger , and A. terreus.

9. The method of any one of claims 1 , 2 , and 4 , wherein the Asp f2-specific antibody or antigen binding fragment thereof comprises the amino acid sequence selected from group consisting of SEQ ID NOs: 1, 2, 20, and 22.

10. A kit for performing the method of any one of claims 1 , 2 , and 4 .

11. An Asp f2-specific antibody or antigen binding fragment thereof comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 1, 2, 20, and 22.

Assignees

• Hope City

Inventors

• Kalkum Markus
• Bagramyan Karine
• Diaz-Arevalo Diana
• Ito James I
• Dadwal Sanjeet

Classifications

• A61P31/10

  Antimycotics · CPC title

• A61K38/04

  Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof ({enzyme inhibitors A61K38/005;} gastrins {A61K38/2207}, somatostatins A61K38/31, melanotropins A61K38/34 {; protease inhibitors A61K38/55}) · CPC title

• A61K38/10

  Peptides having 12 to 20 amino acids {(A61K38/043 - A61K38/046 take precedence)} · CPC title

• A61K39/0002

  Fungal antigens, e.g. Trichophyton, Aspergillus, Candida · CPC title

• G01N2333/962

  from Aspergillus · CPC title