Alkylated interleukin-18 compositions

• US 8,168,165 B2
• Filed: 12/21/2009
• Issued: 05/01/2012
• Est. Priority Date: 12/23/2008
• Status: Expired due to Fees

• Current Assignee: Abbott Laboratories Incorporated
• Original Assignee: Abbott Laboratories Incorporated
• Inventors: Brate, Elaine Marie, Rae, Tracey D., Zhao, Cheng
• Primary Examiner(s): STOICA, ELLY GERALD
• Application Number:
  US12/643,898
• Publication Number:
  US 20100166702A1
• Time in Patent Office: 862 Days
• Field of Search: None
• US Class Current: 424/85.2
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: C07K 14/54 Interleukins [IL]
• CPC Class Codes: Y10T 436/105831 Protein or peptide standard or control [e.g., hemoglobin, etc.]

Abstract

Alkylated interleukin-18 (IL-18), compositions comprising alkylated IL-18, kits comprising such compositions, methods of alkylating IL-18, and methods of using compositions comprising alkylated IL-18.

Claims

1. A composition comprising (i) an isolated or purified, alkylated IL-18, in which at least one free sulfhydryl of a cysteine residue has been alkylated, and (ii) at least one stabilizing protein.

2. The composition of claim 1, in which the hydrogen of at least one free sulfhydryl of a cysteine residue has been replaced with carboxyamidomethyl, and, optionally, in which the protein is bovine serum albumin (BSA).

3. The composition of claim 2, in which the alkylated IL-18 has a capping ratio of about 2.5 to about 4.0 as determined by mass spectrometry, and, optionally, in which the protein is BSA.

4. The composition of claim 1, in which the hydrogen of at least one free sulfhydryl of a cysteine residue has been replaced with carboxymethyl, and, optionally, in which the protein is BSA.

5. The composition of claim 4, in which the alkylated IL-18 has a capping ratio of about 1.0 to about 4.0 as determined by mass spectrometry, and, optionally, in which the protein is BSA.

6. The composition of claim 1, in which the protein is selected from the group consisting of BSA, human serum albumin (HSA), bovine γ

-globulin (BGG), Carnation®

Instant Nonfat Dry Milk, and any combination thereof.

7. The composition of claim 1, in which the protein is BSA.

8. The composition of claim 1, which comprises a known concentration of alkylated IL-18.

9. A series of calibrating compositions, which can be used in a method of determining the concentration of IL-18 in a sample, wherein each of the compositions in the series comprises (i) a known concentration of an isolated or purified, alkylated IL-18, in which at least one free sulfhydryl of a cysteine residue has been alkylated, and (ii) at least one stabilizing protein, wherein each of the compositions differs from the other compositions in the series by the concentration of alkylated IL-18.

10. The series of calibrating compositions of claim 9, in which the hydrogen of at least one free sulfhydryl of a cysteine residue has been replaced with carboxyamidomethyl, and, optionally, in which the protein is BSA.

11. The series of calibrating compositions of claim 10, in which the alkylated IL-18 has a capping ratio of about 2.5 to about 4.0 as determined by mass spectrometry, and, optionally, in which the protein is BSA.

12. The series of calibrating compositions of claim 9, in which the hydrogen of at least one free sulfhydryl of a cysteine residue has been replaced with carboxymethyl, and, optionally, in which the protein is BSA.

13. The series of calibrating compositions of claim 12, in which the alkylated IL-18 has a capping ratio of about 1.0 to about 4.0 as determined by mass spectrometry, and, optionally, in which the protein is BSA.

14. The series of calibrating compositions of claim 9, in which the protein is selected from the group consisting of BSA, HSA, BGG, Carnation®

Instant Nonfat Dry Milk, and any combination thereof.

15. The series of calibrating compositions of claim 9, in which the protein is BSA.

16. A kit for assaying a test sample for IL-18, which kit comprises at least one component for assaying the test sample for IL-18 and instructions for assaying the test sample for IL-18, wherein the at least one component includes at least one composition comprising (i) a known amount of an isolated or purified, alkylated IL-18, in which at least one free sulfhydryl of a cysteine residue has been alkylated, and (ii) at least one stabilizing protein, wherein, if the kit comprises more than one composition, each of the compositions differs from the other compositions by the concentration of alkylated IL-18.