Devices and methods of cell capture and analysis
The present invention provides a device for isolating target biomolecules or cells from samples, particularly biological samples. In particular, the device comprises a loading mixture, which contains the biological sample and a first binding entity that specifically binds to the target biomolecule or target cell; and a micro-channel coated with a second binding entity that binds directly or indirectly to the first binding entity. Methods of capturing, detecting, and/or evaluating target biomolecules or target cells (e.g. cancer cells) in biological samples are also disclosed.
1. A method for evaluating a sample for the presence or absence of a target or biomarker, wherein the method comprises
contacting the sample with a mixture comprising a first binding entity and a third binding entity to allow binding interaction and form a pre-loading mixture, wherein the first binding entity specifically binds to a target entity on the target or biomarker, wherein the first binding entity is an antibody or antibody cocktail that is not conjugated to a detectable or capturable entity, and wherein the third binding entity is an antibody that specifically binds to the first binding entity;
contacting the pre-loading mixture with a surface, wherein the surface is coated with a second binding entity capable of specifically binding to the third binding entity to form a second complex comprising the second binding entity and the first complex,
wherein the second binding entity is a different type of binding molecule than the first binding entity; and
detecting the presence of the target or biomarker on the surface.
2. The method of claim 1 , wherein the target is a rare cell in the biological sample.
3. The method of claim 1 , wherein the method does not involve cytokeratin staining.
4. The method of claim 1 , wherein the target is CK−.
5. The method of claim 1 , wherein the biomarker is aneuploidy of chromosomes 1, 3, 4, 7, 8, 11, 17 and/or 20.
6. The method of claim 1 , wherein the biomarker is monosomy or trisomy 8, 11, and/or 17.
7. The method of claim 1 , wherein the biomarker is monosomy 8, 11, and/or 17.
8. The method of claim 1 , wherein the biomarker is monosomy or tetrasomy 17 or 20.
9. The method of claim 1 , wherein aneuploidy is evaluated by fluorescence in-situ hybridization (FISH).
10. The method of claim 1 , wherein the first binding entity is a primary antibody, the third binding entity is a biotinylated secondary antibody, and wherein the second binding entity is avidin, streptavidin or neutravidin.
11. The method of claim 1 , wherein the sample is obtained from a patient prior to contacting the sample with the first and third binding entity, and wherein the presence or absence of the target or biomarker in the sample is indicative of a condition or disease in the patient.
12. The method of claim 11 , wherein the condition or disease is a cancer, a malignancy, a drug selectivity, or a drug sensitivity.
13. The method of claim 12 , further comprising, detecting an additional marker of malignancy.
14. The method of claim 13 , wherein the additional marker of malignancy is Her2 expression.