MAGNETIC PARTICLE-BASED IMMUNOASSAY AND METHODS OF USING THE SAME

1 . A method for detecting the presence, absence, or level of an analyte of interest in a sample, the method comprising the steps of:

contacting a sample with a magnetic conjugate comprising a magnetic particle and a capture moiety configured to bind the analyte of interest in the sample;

contacting the sample with a reporter conjugate comprising a reporter and a reporter binding moiety configured to bind the analyte of interest in the sample;

binding the analyte of interest with the capture moiety and the reporter binding moiety;

separating the analyte of interest from the sample by applying a magnetic field to the analysis chamber; and

detecting the presence, absence, or level of the analyte of interest by detecting the reporter.

2 . The method of claim 1 , wherein the reporter comprises a metal core and a silica shell.

3 . The method of claim 2 , wherein the silica shell is impregnated with a plurality of quantum dots.

4 . The method of claim 2 or 3 , wherein the metal core comprises gold.

5 . The method of claim 1 , wherein the reporter comprises a plurality of quantum dots.

6 . The method of any one of the preceding claims, wherein the reporter is a fluorescent reporter, a phosphorescent reporter, or a colorimetric reporter.

7 . The method of any one of the preceding claims, further comprising the step of concentrating the analyte of interest in the sample by applying a magnetic field to the analysis chamber after contacting the sample with the magnetic conjugate; and then reducing the volume of the sample in the analysis chamber.

8 . The method of claim 7 , further comprising the step of deactivating the magnetic field before contacting the sample with the reporter conjugate.

9 . The method of any one of the preceding claims, further comprising the steps of concentrating the analyte of interest in the sample by applying a magnetic field to the analysis chamber after contacting the sample with the magnetic conjugate; removing a volume of the sample from the analysis chamber; and adding one or both of a volume of buffer and an additional volume of the sample to the analysis chamber.

10 . The method of claim 9 , further comprising the step of deactivating the magnetic field before contacting the sample with the reporter conjugate.

11 . The method of claim 1 , wherein the reporter conjugate is labeled with biotin and the reporter is functionalized with streptavidin.

12 . The method of any one of the preceding claims, wherein the analyte of interest is selected from the group consisting of human chorionic gonadotropin (hCG), luteinizing hormone (LH)/Lutropin, prostate specific antigen (PSA), herpes simplex virus (HSV) antibodies, estrone-3-glucuronide (E3G), bacteria, hemoglobin A1C, C-reactive protein, an inflammation biomarker, troponin, lyme disease antigen, lyme disease antibodies, an LDL biomarker, an HDL biomarker, a total cholesterol biomarker, thyroid stimulating hormone, a hepatitis C virus biomarker, a rhino virus biomarker, an influenza virus biomarker, a liver function biomarker, estrogen, progesterone, lactic acid, and combinations thereof.

13 . A method for detecting the presence, absence, or level of an analyte of interest in a sample, the method comprising the steps of:

contacting a sample with a magnetic conjugate comprising a magnetic particle and a capture moiety configured to bind the analyte of interest in the sample;

binding the analyte of interest with the capture moiety;

separating the analyte of interest from the sample by applying a magnetic field to the analysis chamber to pull down the magnetic conjugates with analyte of interest associated therewith;

contacting the sample with a reporter conjugate comprising a reporter and a reporter binding moiety configured to bind the analyte of interest in the sample;

binding the analyte of interest with the reporter binding moiety;

separating the analyte of interest with the reporter binding moiety bound thereto from the sample by applying a magnetic field to the analysis chamber; and

detecting the presence, absence, or level of the analyte of interest by detecting the reporter with a light source and photodetector.

14 . The method of claim 13 , wherein the reporter comprises a fluorescent reporter, a phosphorescent reporter, or a colorimetric reporter.

15 . The method of claim 13 , wherein the reporter conjugate comprises a plurality of quantum dots.

16 . The method of any one of claims 13 to 15 , wherein the analyte of interest is selected from the group consisting of human chorionic gonadotropin (hCG), luteinizing hormone (LH)/Lutropin, prostate specific antigen (PSA), herpes simplex virus (HSV) antibodies, estrone-3-glucuronide (E3G), bacteria, hemoglobin A1C, C-reactive protein, an inflammation biomarker, troponin, lyme disease antigen, lyme disease antibodies, an LDL biomarker, an HDL biomarker, a total cholesterol biomarker, thyroid stimulating hormone, a hepatitis C virus biomarker, a rhino virus biomarker, an influenza virus biomarker, a liver function biomarker, estrogen, progesterone, lactic acid, and combinations thereof.

17 . A method for detecting the presence, absence, or level of an analyte of interest in a sample, the method comprising the steps of:

contacting a sample with a magnetic conjugate comprising a magnetic particle and a capture moiety configured to bind the analyte of interest in the sample;

binding the analyte of interest with the capture moiety;

contacting the sample with a reporter-labeled analyte configured to bind the magnetic conjugate in the absence of the analyte of interest in the sample;

separating the analyte of interest from the sample by applying a magnetic field to the sample; and

detecting the presence, absence, or level of the analyte of interest by detecting the reporter.

18 . The method of claim 17 , wherein the reporter comprises a fluorescent reporter, a phosphorescent reporter, or a colorimetric reporter.

19 . The method of claim 17 or 18 , wherein reporter-labeled analyte comprises a plurality of quantum dots.

20 . The method of any one of claims 17 to 19 , wherein the analyte of interest is selected from the group consisting of human chorionic gonadotropin (hCG), luteinizing hormone (LH)/Lutropin, prostate specific antigen (PSA), herpes simplex virus (HSV) antibodies, estrone-3-glucuronide (E3G), bacteria, hemoglobin A1C, C-reactive protein, an inflammation biomarker, troponin, lyme disease antigen, lyme disease antibodies, an LDL biomarker, an HDL biomarker, a total cholesterol biomarker, thyroid stimulating hormone, a hepatitis C virus biomarker, a rhino virus biomarker, an influenza virus biomarker, a liver function biomarker, estrogen, progesterone, lactic acid, and combinations thereof.

21 . A method for detecting the presence, absence, or level of an analyte of interest in a sample, the method comprising the steps of:

contacting a sample with a reporter conjugate comprising a reporter and a reporter binding moiety configured to bind the analyte of interest in the sample;

binding the analyte of interest with the reporter binding moiety;

contacting the sample with a magnetic particle-labeled analyte configured to bind the reporter conjugate in the absence of the analyte of interest in the sample;

separating the magnetic particle-labeled analyte from the sample by applying a magnetic field to the sample; and

detecting the presence, absence, or level of the analyte of interest by detecting the reporter.

22 . The method of claim 21 , wherein the reporter comprises a fluorescent reporter, a phosphorescent reporter, or a colorimetric reporter.

23 . The method of claim 21 or 22 , wherein the reporter conjugate comprises a plurality of quantum dots.

24 . The method of any one of claims 21 to 23 , wherein the analyte of interest is selected from the group consisting of human chorionic gonadotropin (hCG), luteinizing hormone (LH)/Lutropin, prostate specific antigen (PSA), herpes simplex virus (HSV) antibodies, estrone-3-glucuronide (E3G), bacteria, hemoglobin A1C, C-reactive protein, an inflammation biomarker, troponin, lyme disease antigen, lyme disease antibodies, an LDL biomarker, an HDL biomarker, a total cholesterol biomarker, thyroid stimulating hormone, a hepatitis C virus biomarker, a rhino virus biomarker, an influenza virus biomarker, a liver function biomarker, estrogen, progesterone, lactic acid, and combinations thereof.

25 . A method for detecting the presence, absence, or level of an analyte of interest in a sample, the method comprising the steps of:

contacting a sample with a magnetic conjugate comprising a magnetic particle and a capture moiety configured to bind the analyte of interest in the sample;

binding the analyte of interest with the capture moiety;

contacting the sample with a reporter binding moiety comprising a biotin label configured to bind the analyte of interest in the sample;

contacting the sample with a reporter comprising a streptavidin label configured to bind the biotin label;

separating the analyte of interest from the sample by applying a magnetic field to the sample; and

detecting the presence, absence, or level of the analyte of interest by detecting the reporter.

26 . The method of claim 25 , wherein the reporter comprises a fluorescent reporter, a phosphorescent reporter, or a colorimetric reporter.

27 . The method of claim 25 or 26 , wherein the reporter comprises a plurality of quantum dots.

28 . The method of any one of claims 25 to 27 , wherein the analyte of interest is selected from the group consisting of human chorionic gonadotropin (hCG), luteinizing hormone (LH)/Lutropin, prostate specific antigen (PSA), herpes simplex virus (HSV) antibodies, estrone-3-glucuronide (E3G), bacteria, hemoglobin A1C, C-reactive protein, an inflammation biomarker, troponin, lyme disease antigen, lyme disease antibodies, an LDL biomarker, an HDL biomarker, a total cholesterol biomarker, thyroid stimulating hormone, a hepatitis C virus biomarker, a rhino virus biomarker, an influenza virus biomarker, a liver function biomarker, estrogen, progesterone, lactic acid, and combinations thereof.

29 . The method of any one of claims 1 to 28 , comprising the step of adding the sample to an analysis chamber.

30 . The method of claim 29 , wherein the step of contacting the sample with the magnetic conjugate comprises contacting the sample with the magnetic conjugate in the analysis chamber.

31 . The method of claim 29 or 30 , wherein the step of adding the sample to the analysis chamber comprises adding the sample to a sample collector in fluid communication with the analysis chamber.

32 . The method of claim 31 , wherein the step of contacting the sample with the magnetic conjugate comprises contacting the sample with the magnetic conjugate in the sample collector.

33 . A kit for providing the method of any one of claims 1 to 32 .

34 . A method for evaluating a biological event comprising measuring a biomarker using the method of any one of claims 1 to 32 .