Compositions, kits, and methods for cell separation
A cell-capturing substrate, methods of using the cell-capturing substrate that allow for label-free cell separation, and kits that incorporate the cell-capturing substrate.
1. A cell separation substrate comprising:
a substrate;
a capture antibody bound to the substrate, wherein the capture antibody is selected to bind to a target cell by binding to a cell surface protein expressed by the target cell;
an effector bound to the substrate, the effector comprising a protein comprising a coiled coil domain; and
a molecular trigger comprising:
a protein comprising a coiled coil domain that forms a heterodimeric complex with the coiled coil domain of the effector, and
a domain comprising a hydrophilic polymer chain having a molecular weight of at least 5 kDa;
wherein the effector is bound to the substrate in proximity to the capture antibody so that the heterodimer complex disrupts binding between the capture antibody and the target cell.
2. The cell separation substrate of claim 1 wherein the substrate comprises a tissue culture plate.
3. The cell separation substrate of claim 1 wherein the substrate comprises a chip, a biochip, a bead, a magnetic bead, a microarray, a microfluidic channel, or a microfluidic chamber.
4. The cell separation substrate of claim 1 wherein the capture antibody comprises an antibody or an antibody fragment.
5. The cell separation substrate of claim 1 wherein the capture antibody comprises an anti-CD31 antibody, an anti-CD3 antibody, an anti-CD4 antibody, an anti-CD8 antibody, an anti-CD19 antibody, an anti-CD20 antibody, an anti-CD34 antibody, an anti-CD56 antibody, an anti-CD235a antibody, an anti-CD146 antibody, an anti-CD326 antibody, an anti-CD11 c antibody, an anti-CD123 antibody, an anti-CD14 antibody, an anti-CD33 antibody, an anti-CD66b antibody, an anti-CD41 antibody, an anti-CD61 antibody, an anti-CD62, an anti-EpCAM antibody, an anti-N-cadherin antibody, an anti-CD44 antibody, an anti-CD24 antibody, an anti-CD133 antibody, an anti-CD166 antibody, an anti-CD134 antibody, an anti-CD137 antibody, an anti-CD28 antibody, an anti-CD25 antibody, an anti-CD27 antibody, an anti-PD-1 antibody, an anti-SCA1 antibody, an anti-IL-2Rβ antibody, an anti-CD45RA antibody, an anti-CD62L antibody, or a mixture thereof.
6. The cell separation substrate of claim 1 wherein the heterodimerizable complex comprises a reversible, noncovalent complex comprising self-assembly of the effector and the molecular trigger.
7. The cell separation substrate of claim 1 , wherein the effector comprises SEQ ID NO: 1.
8. The cell separation substrate of claim 1 , wherein the molecular trigger comprises SEQ ID NO:2.
9. A cell-capturing substrate comprising:
a substrate;
a capture antibody bound to the substrate, the capture antibody being selected to bind to a target cell by binding to a cell surface protein expressed by the target cell;
a target cell bound to the capture antibody;
an effector bound to the substrate, the effector comprising a protein comprising a coiled coil domain; and
a molecular trigger comprising:
a protein comprising a coiled coil domain that forms a heterodimeric complex with the coiled coil of the effector, and
a domain comprising a hydrophilic polymer chain having a molecular weight of at least 5 kDa;
wherein the effector is bound to the substrate in proximity to the capture antibody so that the heterodimer complex disrupts binding between the capture antibody and the target cell.
10. The cell separation substrate of claim 1 , wherein the hydrophilic polymer chain of the molecular trigger has a molecular weight of at least 10 kDa.
11. The cell-capturing substrate of claim 9 , wherein the hydrophilic polymer chain of the molecular trigger has a molecular weight of at least 10 kDa.