Devices and methods for detecting cells and other analytes
The invention features methods, devices, and kits for the isolation of analytes (e.g., a cell). A sample containing a desired analyte is introduced into a microfluidic device containing moieties that bind the desired analyte. A shear stress is applied that is great enough to prevent binding of undesired analytes and low enough to allow binding of the analyte of interest. Once bound, the desired analytes can be analyzed (e.g., counted). The invention also features methods for determining a shear stress for isolating a desired analyte.
1. A method for isolating CD4+ lymphocytes from a sample of whole blood, the method comprising:
(a) introducing said sample into a microfluidic device coated with binding moieties that bind to a cell surface marker of said CD4+ lymphocytes; and
(b) applying a controlled shear stress from greater than 0.5 dyn/cm 2 to less than about 7.0 dyn/cm 2 to said sample in the microfluidic device, wherein the shear stress is low enough to allow binding of the binding moieties to said CD4+ lymphocytes expressing the cell surface marker at a first concentration and great enough to inhibit binding of the binding moieties to undesired CD4+ monocytes that express the same cell surface marker at a second concentration lower than the first concentration.
2. The method of claim 1 , wherein said binding moieties are anti-CD4 binding moieties selected from the group consisting of antibodies, antibody fragments, oligopeptides, polypeptides, aptamers, oligonucleotides, coordination complexes, synthetic polymers, and carbohydrates.
3. The method of claim 2 , wherein said sample is obtained from a patient at risk of developing AIDS.
4. The method of claim 1 , further comprising (c) analyzing at least one biological property of said CD4+ lymphocytes.
5. The method of claim 4 , wherein said biological property is selected from the group consisting of: mRNA expression, protein expression, DNA quantification, DNA sequence, and chromosomal abnormalities.
6. The method of claim 2 , further comprising: (c) counting said CD4+ lymphocytes that bind to said binding moieties.
7. The method of claim 6 , further comprising using said count to diagnose a disease state.
8. The method of claim 1 , wherein binding of said CD4+ lymphocytes to said binding moieties occurs simultaneously with the application of the controlled shear stress.
9. The method of claim 1 , wherein the controlled shear stress is between about 1 dyn/cm 2 and about 3 dyn/cm 2 .
10. The method of claim 1 , wherein the controlled shear stress is between about 0.7 dyn/cm 2 and about 5.0 dyn/cm 2 .
11. The method of claim 1 , wherein the controlled shear stress is between about 0.7 dyn/cm 2 and about 4.0 dyn/cm 2 .