Methods and apparatus for magnetic separation of cells
First Claim
1. A method for capturing and isolating cells comprising:
- (a) mixing a sample comprising rare target cells and contaminant cells with magnetically responsive particles having an affinity to the rare target cells to produce a sample solution having magnetically responsive particles bound to rare target cells;
(b) contacting a magnetic member having a non-adherent sleeve with the sample solution;
(c) producing a continuous relative motion between the magnetic member and the sample solution while the magnetic member produces a magnetic field across the non-adherent sleeve, such that the magnetically responsive particles bound to rare target cells are selectively captured onto the non-adherent sleeve and wherein the continuous relative motion is applied in concentric circles such that a majority of the sample solution has access to the magnetic member;
(d) contacting the magnetic member having the captured rare target cells with a recovery solution;
(e) substantially removing the magnetic field produced by the member across the non-adherent sleeve whereby the magnetically responsive particles bound to rare target cells are released into the recovery fluid.
2 Assignments
0 Petitions
Accused Products
Abstract
Described here is an automated robotic device that isolates circulating tumor cells (CTCs) or other biological structures with extremely high purity. The device uses powerful magnetic rods covered in removable plastic sleeves. These rods sweep through blood samples, capturing, e.g., cancer cells labeled with antibodies linked to magnetically responsive particles such as superparamagnetic beads. Upon completion of the capturing protocol, the magnetic rods undergo several rounds of washing, thereby removing all contaminating blood cells. The captured target cells are released into a final capture solution by removing the magnetic rods from the sleeves. Additionally, cells captured by this device show no reduced viability when cultured after capture. Cells are captured in a state suitable for genetic analysis. Also disclosed are methods for single cell analysis. Being robotic allows the device to be operated with high throughput.
79 Citations
41 Claims
-
1. A method for capturing and isolating cells comprising:
-
(a) mixing a sample comprising rare target cells and contaminant cells with magnetically responsive particles having an affinity to the rare target cells to produce a sample solution having magnetically responsive particles bound to rare target cells; (b) contacting a magnetic member having a non-adherent sleeve with the sample solution; (c) producing a continuous relative motion between the magnetic member and the sample solution while the magnetic member produces a magnetic field across the non-adherent sleeve, such that the magnetically responsive particles bound to rare target cells are selectively captured onto the non-adherent sleeve and wherein the continuous relative motion is applied in concentric circles such that a majority of the sample solution has access to the magnetic member; (d) contacting the magnetic member having the captured rare target cells with a recovery solution; (e) substantially removing the magnetic field produced by the member across the non-adherent sleeve whereby the magnetically responsive particles bound to rare target cells are released into the recovery fluid. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 27)
-
-
13. A method for capture and isolation of intact, rare target cells in a sample having a mixed cell population of target cells and contaminant cells, wherein the target cells are labeled with magnetically responsive material to form labeled target cells, comprising:
-
(a) labeling the sample with a label specific for target cells, said label being magnetically responsive, to form a labeled sample; (b) contacting the labeled sample with a member extending into the sample and comprising a strong magnet covered by a nonmagnetic, nonadherent sleeve; (c) sweeping the member in a continuous motion through the sample to cause cells to attach to the sleeve wherein the sweeping is orbital; (d) washing the sleeve with cells attached to remove unlabeled cells into a cell free solution; (e) separating the magnet from the sleeve, whereby labeled cells are removed from the sleeve; and (f) collecting labeled cells to form a composition comprising beads and labeled cells. - View Dependent Claims (14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
-
-
28. A method for capturing and isolating cells comprising:
-
(a) mixing a sample comprising rare target cells and contaminant cells with magnetically responsive particles having an affinity to the rare target cells to produce a sample solution having magnetically responsive particles bound to rare target cells; (b) contacting a magnetic member having a non-adherent sleeve with the sample solution; (c) producing a continuous relative motion between the magnetic member and the sample solution while the magnetic member produces a magnetic field across the non-adherent sleeve, such that the magnetically responsive particles bound to rare target cells are selectively captured onto the non-adherent sleeve and wherein said continuous relative motion is orbital; (d) contacting the magnetic member having the captured rare target cells with a recovery solution; (e) substantially removing the magnetic field produced by the member across the non-adherent sleeve whereby the magnetically responsive particles bound to rare target cells are released into the recovery fluid. - View Dependent Claims (29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41)
-
Specification