Methods for detecting pre-diabetes and diabetes using differential protein glycosylation

• US 8,497,077 B2
• Filed: 04/26/2012
• Issued: 07/30/2013
• Est. Priority Date: 05/11/2009
• Status: Active Grant

• Current Assignee: DiabetOmics, Inc.
• Original Assignee: Diabetomics, LLC
• Inventors: Nagalla, Srinivasa R., Roberts, Charles T.
• Primary Examiner(s): Cheu, Jacob
• Application Number:
  US13/457,243
• Publication Number:
  US 20120208218A1
• Time in Patent Office: 460 Days
• Field of Search: None
• US Class Current: 435/7.1
• CPC Class Codes: G01N 2333/42 Lectins, e.g. concanavalin, phytohaemagglutinin
• CPC Class Codes: G01N 2400/00 Assays, e.g. immunoassays or enzyme assays, involving carbohydrates
• CPC Class Codes: G01N 2440/38 addition of carbohydrates, e.g. glycosylation, glycation
• CPC Class Codes: G01N 2500/02 Screening involving studying the effect of compounds C on the interaction between interacting molecules A and B (e.g. A = enzyme and B = substrate for A, or A = receptor and B = ligand for the receptor)
• CPC Class Codes: G01N 2800/042 Disorders of carbohydrate metabolism, e.g. diabetes, glucose metabolism
• CPC Class Codes: G01N 2800/50 Determining the risk of developing a disease
• 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 33/6893 related to diseases not provided for elsewhere

Abstract

Methods for identifying individuals who are not yet diabetic (pre-diabetic), but who are at significant risk of developing diabetes, such as type 2 diabetes, are disclosed herein. Methods are also provided for the identification of diabetic subjects. Also disclosed are methods for identifying individuals with diabetic complications. The methods include the identification of an overall glycosylation profile of proteins in a biological fluid, such as saliva, urine, or serum. In some examples, the methods include determining the amount of one or more protein in a biological fluid or determining the glycosylation pattern of one or more proteins in a biological fluid.

Claims

1. A lectin-binding test device for determining whether a subject has diabetes or pre-diabetes, wherein the lectin-binding test device comprises:

a plurality of lectins, wherein the plurality of lectins comprises Aleuria aurantia lectin, Datura stramonium lectin, and Phaseolus vulgaris agglutinin;

a support for the plurality of lectins; and

an indicator to detect binding of each of the plurality of lectins to a glycosylated protein in the sample, wherein an increase in binding of Aleuria aurantia lectin, Datura stramonium lectin, and Phaseolus vulgaris agglutinin relative to a non-diabetic control value indicates that the subject has diabetes or pre-diabetes.

2. The lectin binding test device of claim 1, wherein the plurality of lectins further comprises Concanavalin A and Sambucus nigra lectin, and wherein an increase in binding of Aleuria aurantia lectin, Datura stramonium lectin, non-diabetic control indicates that the subject has diabetes, and not pre-diabetes.

3. The lectin binding test device of claim 1, wherein the plurality of lectins consists of Aleuria aurantia lectin, Datura stramonium lectin, and Phaseolus vulgaris agglutinin.

4. The lectin binding test device of claim 2, wherein the plurality of lectins consists of Aleuria aurantia lectin, Datum stramonium lectin. Phaseolus vulgaris agglutinin, Concanavalin A, and Sambucus nigra lectin.

5. The lectin binding test device of claim 2, wherein the test device provides a quantitative measure of binding for each of the plurality of lectins.

6. The lectin binding test device of claim 5, wherein the test device provides a ratio of binding for at least two of the plurality of lectins.

7. The lectin binding test device of claim 6, wherein the plurality of lectins further comprises Maackia amurensis lectin I.

8. The lectin binding test device of claim 7, wherein an increase in a ratio of binding of Sambucus nigra lectin to Maackia amurensis lectin I, Concanavalin A to Maackia amurensis lectin I, or Concanavalin A to Aleuria aurantia lectin indicates that the sample is from a subject having diabetes.

9. The lectin binding test device of claim 1, wherein the device further comprises an anti-alpha-1 acid glycoprotein (A1AG) antibody or an anti-alpha-1 antitrypsin (A1 AT) antibody.

10. The lectin binding test device of claim 9, wherein the anti-alpha-1 acid glycoprotein (A1AG) antibody and/or anti-alpha-1 antitrypsin (A1AT) antibody provides a quantitative measure of an amount of alpha-1 acid glycoprotein (A1AG) and/or alpha-1 antitrypsin (A1AT) protein in the sample.

11. The lectin binding test device of claim 10, wherein an increase in the quantity of binding of A1AG and/or A1AT as compared to a control value indicates that the subject has pre-diabetes or diabetes.

12. The lectin binding test device of claim 1, wherein at least one of the plurality of lectins binds alpha-1 acid glycoprotein.

13. The lectin binding test device of claim 2, wherein the plurality of lectins further comprises Maackia amurensis lectin I, and wherein an increase in a ratio of binding of Sambucus nigra lectin to Maackia amurensis lectin I indicates that the sample is from a subject having diabetes.

14. The lectin binding test device of claim 1, wherein the plurality of lectins further comprises Maackia amurensis lectin I, and wherein an increase in a ratio of binding of Concanavalin A to Maackia amurensis lectin I indicates that the sample is from a subject having diabetes.

15. The lectin binding test device of claim 2, wherein an increase in a ratio of binding of Concanavalin A to Aleuria aurantia lectin indicates that the sample is from a subject having diabetes.