SPECTROSCOPIC METHODS FOR THE DETECTION OF GLYCATED HEMOGLOBIN

• US 20170030932A1
• Filed: 04/10/2015
• Published: 02/02/2017
• Est. Priority Date: 04/11/2014
• Status: Active Application

• Current Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Original Assignee: Siemens Healthcare Diagnostics Incorporated (Siemens AG)
• Inventors: Ledden, David J.
• Granted Patent: US 10,288,628 B2
• Time in Patent Office: Days
• US Class Current: 1/1
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/10 Integrating sample preparation and analysis in single entity, e.g. lab-on-a-chip concept
• CPC Class Codes: B01L 2300/0627 Sensor or part of a sensor is integrated
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/087 Multiple sequential chambers
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: G01N 33/491 by separating the blood components G01N15/05 takes precedence
• CPC Class Codes: G01N 33/52 Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements immunological elements G01N33/54386; involving labelled immunochemicals G01N33/58; for haemoglobin or occult blood G01N33/72
• CPC Class Codes: G01N 33/721 Haemoglobin
• CPC Class Codes: G01N 33/723 Glycosylated haemoglobin
• CPC Class Codes: Y10T 436/255 including use of a solid sorbent, semipermeable membrane, or liquid extraction

Abstract

Kits, microfluidics devices, and assays for use in methods of spectroscopically determining a ratio of glycated hemoglobin to total hemoglobin in a whole blood sample are disclosed.

Claims

1. A kit for determining a ratio of glycated hemoglobin to total hemoglobin, the kit comprising:

a blood separation membrane, wherein the blood separation membrane traps red blood cells thereon and/or therein, and wherein plasma passes through the blood separation membrane;

a waste collection material capable of being associated with the blood separation membrane and capable of wicking waste material away from the blood separation membrane and into the waste collection material;

a negatively charged surface, wherein the negatively charged surface traps hemoglobin thereon and/or therein, and wherein other components of lysed red blood cells are not trapped on the negatively charged surface, and wherein the negatively charged surface is capable of being subjected to spectroscopy to measure an amount of total hemoglobin trapped thereon and/or therein and an amount of glycated hemoglobin or non-glycated hemoglobin trapped thereon and/or therein; and

a waste collection material capable of being associated with the negatively charged surface and capable of wicking waste material away from the negatively charged surface and into the waste collection material.

2. The kit of claim 1, wherein the negatively charged surface is capable of being subjected to at least one of near-infrared (NIR) spectroscopy, mid-infrared (MIR) spectroscopy, and visible spectroscopy.

3. The kit of claim 2, further comprising a labeling agent capable of labeling either glycated hemoglobin or non-glycated hemoglobin.

4. The kit of claim 3, wherein the labeling agent comprises boronate, wherein the boronate is capable of labeling glycated hemoglobin.

5. The kit of claim 4, wherein the boronate is labeled with a NIR/MIR dye.

6. A microfluidics device for determining a ratio of glycated hemoglobin to total hemoglobin, the microfluidics device comprising:

a first compartment capable of receiving a whole blood sample, the at least one compartment comprising a blood separation membrane and a waste collection material positioned therebelow and associated therewith, wherein the blood separation membrane traps red blood cells thereon and/or therein, and wherein plasma passes through the blood separation membrane and is wicked away from the blood separation membrane and into the waste collection material;

a second compartment comprising a negatively charged surface and a waste collection material positioned therebelow and associated therewith, wherein the negatively charged surface traps hemoglobin thereon and/or therein, and wherein other components of lysed red blood cells are not trapped on the negatively charged surface and are wicked away from the negatively charged surface and into the waste collection material, and wherein the negatively charged surface is capable of being subjected to spectroscopy to measure an amount of total hemoglobin trapped thereon and/or therein and an amount of glycated hemoglobin or non-glycated hemoglobin trapped thereon and/or therein;

an optical read chamber capable of being optically interrogated by a spectrometer;

wherein the blood separation membrane is capable of releasable movement from association with the waste collection material to association with the negatively charged surface in the second compartment; and

wherein the negatively charged surface is capable of releasable movement from association with the blood separation membrane to a position within the optical read chamber, and wherein the negatively charged surface can be subjected to spectroscopy within the optical read chamber of the third compartment for measuring (1) an amount of total hemoglobin present on the negatively charged surface, and (2) an amount of one of glycated hemoglobin or non-glycated hemoglobin present on the negatively charged surface.

7. The microfluidics device of claim 6, further comprising:

a first actuator for releasably moving the blood separation membrane movement from association with the waste collection material to association with the negatively charged surface in the second compartment; and

a second actuator for releasably moving the negatively charged surface from association with the waste collection material to a position within the optical read chamber.

8. The microfluidics device of claim 6, wherein the negatively charged surface is capable of being subjected to at least one of near-infrared (NIR) spectroscopy, mid-infrared (MIR) spectroscopy, and visible spectroscopy within the optical read chamber.

9. The microfluidics device of claim 8, wherein the labeling agent comprises boronate, wherein the boronate labels one of glycated hemoglobin or non-glycated hemoglobin on and/or in the negatively charged surface, wherein the boronate is labeled with a NIR/MIR dye.

10. A method of determining a ratio of glycated hemoglobin to total hemoglobin in a whole blood sample, the method comprising the steps of:

applying a whole blood sample on an upper surface of a blood separation membrane of a microfluidics device, the blood separation membrane having a lower surface that is associated with a first waste collection material, whereby red blood cells in the whole blood sample are captured on and/or in the blood separation membrane while plasma passes through the blood separation membrane and is wicked away from the blood separation membrane and into the first waste collection material;

moving a portion of the microfluidics device so that the blood separation membrane is positioned over a negatively charged surface having a second waste collection material positioned therebelow and in association therewith;

applying a lysing agent to the blood separation membrane, wherein the contents of the lysed red blood cells pass through the blood separation membrane and are brought into contact with the negatively charged surface, and wherein the negatively charged surface traps hemoglobin thereon and/or therein while the other components of lysed red blood cells are wicked away from the negatively charged surface and into the second waste collection material;

moving a portion of the microfluidics device so that the negatively charged surface is positioned within an optical read chamber that allows for optical interrogation of the negatively charged surface by a spectrometer;

spectroscopically measuring an amount of total hemoglobin on and/or in the negatively charged surface and an amount of one of glycated hemoglobin or non-glycated hemoglobin on and/or in the negatively charged surface; and

determining a ratio of glycated hemoglobin to total hemoglobin in the whole blood sample based on the spectroscopic measurements.

11. The method of claim 10, wherein the negatively charged surface is spectroscopically measured by at least one of near-infrared (NIR) spectroscopy, mid-infrared (MIR) spectroscopy, and visible spectroscopy.

12. The method of any of claims 11, wherein the labeling agent is boronate, wherein the boronate labels glycated hemoglobin trapped on and/or in the negatively charged surface, wherein the boronate is labeled with a NIR/MIR dye.

13. A method of determining a ratio of glycated hemoglobin to total hemoglobin in a whole blood sample, the method comprising the steps of:

applying a whole blood sample on an upper surface of a blood separation membrane of a microfluidics device, the blood separation membrane having a lower surface that is associated with a waste collection material, whereby red blood cells in the whole blood sample are captured on and/or in the blood separation membrane while plasma passes through the blood separation membrane and is wicked away from the blood separation membrane and into the waste collection material;

applying a labeling agent to the blood separation membrane, wherein the labeling agent penetrates the red blood cells and labels either glycated hemoglobin or non-glycated hemoglobin, and wherein excess labeling agent that did not penetrate the red blood cells is wicked away from the blood separation membrane and into the waste collection material;

applying a wash solution to the blood separation membrane, wherein the wash solution is wicked away from the blood separation membrane and into the waste collection material; and

measuring, via at least one of near-infrared (NIR) spectroscopy, mid-infrared (MIR) spectroscopy, and visible spectroscopy;

(i) an amount of total hemoglobin on the blood separation membrane, and

(ii) an amount of either;

(a) labeled glycated hemoglobin on the blood separation membrane, or

(b) labeled non-glycated hemoglobin on the blood separation membrane; and

determining a ratio of glycated hemoglobin to total hemoglobin in the whole blood sample based on the spectroscopic measurements.

14. The method of claim 13, wherein the labeling agent is boronate, wherein the boronate labels glycated hemoglobin on and/or in the blood separation membrane.

15. The method of claim 14, wherein the boronate is labeled with a NIR/MIR dye.