Cellular controls for glycated hemoglobin Hb A1c

• US 7,361,513 B2
• Filed: 04/08/2005
• Issued: 04/22/2008
• Est. Priority Date: 04/08/2005
• Status: Expired due to Fees

• Current Assignee: Streck
• Original Assignee: Streck
• Inventors: Wu, Jiong, Ryan, Wayne L.
• Primary Examiner(s): WALLENHORST, MAUREEN
• Application Number:
  US11/102,378
• Publication Number:
  US 20060228803A1
• Time in Patent Office: 1,110 Days
• Field of Search: 436/8, 436/15, 436/18, 436/63, 436/66, 436/67, 436/174, 436/175, 252/408.1, 435/2
• US Class Current: 436/67
• CPC Class Codes: G01N 33/723 Glycosylated haemoglobin
• CPC Class Codes: Y10T 436/10 Composition for standardization, calibration, simulation, stabilization, preparation or preservation; processes of use in preparation for chemical testing
• CPC Class Codes: Y10T 436/105831 Protein or peptide standard or control [e.g., hemoglobin, etc.]
• CPC Class Codes: Y10T 436/108331 Preservative, buffer, anticoagulant or diluent
• CPC Class Codes: Y10T 436/25 including sample preparation
• CPC Class Codes: Y10T 436/25125 Digestion or removing interfering materials

Abstract

Disclosed are cellular hemoglobin A1c (Hb A1c) normal and abnormal (high) controls for use in detecting Hb A1c levels. The present invention also relates to methods for generating cellular Hb A1c controls using red blood cells and methods for using the cellular controls. The present invention encompasses several methods for the preparation of Hb A1c cellular controls including: (1) a boronate method where the glycation occurs non-specifically; (2) a stabilized diabetic blood method where the glycation occurs specifically on Hb A1c, and (3) the glycation of normal blood method that is achieved by controlling conditions such that glycation occurs predominantly on Hb A1c. These methods produce cellular Hb A1c controls with desirable stability and that can be detected on a variety of instruments.

Claims

1. A method for preparing a cellular hemoglobin A1c (Hb A1c) control comprising:

(a) selecting a sample of red blood cells with at least one desired feature from a suitable subject;

(b) washing the sample;

(c) processing the washed sample to remove white blood cells;

(d) preserving Hb A1c molecules in said sample, wherein said red blood cells are intact and said Hb A1c molecules are unmodified;

(e) washing the sample of (d); and

(f) adjusting the Hb A1c level to the predetermined level thereby producing a cellular Hb A1c control, wherein the level of Hb A1c of the control is substantially stabilized and detectable.

2. The method of claim 1, further comprising:

(g) admixing the control of (f) in a suspension medium suitable for delivering said control to a suitable analytical instrument for analysis.

3. The method of claim 1, wherein said suitable subject is a mammal, an avian, or a reptile subject.

4. The method of claim 3, wherein said sample is obtained from human, bovine, or both human and bovine subjects.

5. The method of claim 3, wherein said sample is obtained from a diabetic subject.

6. The method of claim 1, wherein the Hb A1c level of the control is detectable using immunologic detection, ion exchange, or affinity chromatography.

7. The method of claim 1, wherein the Hb A1c level of the control is about 1-5%.

8. The method of claim 1, wherein the Hb A1c level of the control is greater than 5%.

9. The method of claim 1, wherein the Hb A1c level of the control is about 5-20%.

10. The method of claim 1, wherein said preserving comprises treating the cells with about 0.00 1-3% polyethylene glycol.

11. The method of claim 10, wherein said preserving comprises treating the cells with about 0.001-1% glucose.

12. The method of claim 1, wherein said preserving comprises fixing the red blood cells.

13. The method of claim 12, wherein said fixing comprises treating the cells with about 0.002-0.10% glutaraldehyde.

14. The method of claim 1, wherein said adjusting comprises maintaining pH at about 6-8.

15. The method of claim 1, further comprising after (c), incubating the red blood cells with about 3% glucose and about 0.5% NaCNBH₃ at room temperature.

16. The method of claim 1, further comprising after (c), incubating the red blood cells for at least about 50 days, at about 4-6°

C., with a glycation solution which comprises;

0.04% Methyl paraben;

0.3% PEG 20K;

0.025% Inosine;

0.015% Chioramphenicol

0.04% Neomycin Sulfate

0.585% EDTA (2Na⁺);

0.325% Magnesium gluconate;

0.225% Na₂HPO₄;

0.07% NaOH;

0.005% NaF; and

3.15% Glucose;

wherein the osmolality of the glycation solution is adjusted to about 300±

7 (mOsm) and pH is adjusted to about 7.0±

0.1.

17. The method of claim 1, wherein said red blood cells are unlysed.

18. A cellular hematology control for Hb A1c comprising intact red blood cells having a predetermined level of Hb A1c, wherein the Hb A1c is unmodified, and the level is substantially stabilized.

19. The control of claim 18, wherein the red blood cells have a substantially stable Hb A1c content for at least about one month.

20. The control of claim 18, wherein the red blood cells have a substantially stable Hb A1c content for at least about three months.

21. The control of claim 18, wherein the Hb A1c level is about 1-5%.

22. The control of claim 18, wherein the Hb A1c level is greater than 5%.

23. The control of claim 18, wherein the Hb A1c level is about 5-20%.

24. The control of claim 18, wherein the predetermined level of Hb A1c is stabilized through adjusting pH of the control to about 6-8.

25. The control of claim 18, wherein the predetermined level of Hb A1c is stabilized through the addition of glucose at an amount of about 0.001-1%.

26. A method for determining the accuracy and reproducibility of the operation of an analytical instrument capable of measuring Hb A1c levels comprising

(a) providing the cellular hematology control for Hb A1c of claim 18 in a known reference quantity;

(b) determining the Hb A1c level in said control of (a) with said instrument; and

(c) comparing the Hb A1c level obtained in (b) with said known reference quantity;

wherein said comparing indicates the accuracy and reproducibility of the operation of said analytical instrument.

27. The control of claim 18, wherein said red blood cells are unlysed.