Methods and Devices for Capturing Circulating Tumor Cells

Abstract

A method of capturing a Circulating Tumor Cell (CTC) from a sample includes introducing a sample into a microfluidic device having a cell capture surface and a flow modification surface under conditions that allow a CTC to bind to a cell rolling-inducing agent and a capturing agent disposed on the cell capture surface. The flow modification surface induces a rotational flow within the sample as it flows through the microfluidic device.

1 . A method of capturing a Circulating Tumor Cell (CTC) from a sample comprising the step of introducing said sample into a microfluidic device under conditions that allow a CTC to bind to a cell rolling-inducing agent and a capturing agent, the device inducing a rotational flow with the sample, the device comprising an immobilized cell rolling-inducing agent and an immobilized capturing agent. 
     
     
         2 . The method of  claim 1  further comprising applying a shear stress between 0.05 and 10 dyn/cm 2  on the sample introduced into the device. 
     
     
         3 . The method of  claim 2  wherein the shear stress is between 0.1 and 2.0 dyn/cm 2 . 
     
     
         4 . The method of  claim 3  wherein the shear stress is about 0.16 dyn/cm 2 . 
     
     
         5 . The method of  claim 1  wherein the cell rolling-inducing agent is a selectin or a CTC binding fragment of a selectin. 
     
     
         6 . The method of  claim 5  wherein the selectin is selected from the group consisting of E-selectin, P-selectin, and L-selectin. 
     
     
         7 . The method of  claim 1  wherein the capturing agent specifically binds a moiety on a CTC cell surface, the capturing agent selected from the group consisting of an antibody, an antibody fragment, an engineered antibody, folic acid, transferrin, a peptide, and an aptamer. 
     
     
         8 . The method of  claim 7  wherein the antibody is anti-EpCAM. 
     
     
         9 . The method of  claim 7  wherein the peptide is an RGD peptide. 
     
     
         10 - 22 . (canceled) 
     
     
         23 . A microfluidic device for capturing a Circulating Tumor Cell (CTC) from a sample, comprising:
 a channel comprising a cell capture surface and a flow modification surface,   wherein the cell capture surface comprises a cell rolling-inducing agent and a capturing agent immobilized on the cell capture surface, and   the flow modification surface comprises one or more ridges extending into the channel and arranged to induce a rotational flow in a sample flowing through the channel.   
     
     
         24 . The device of  claim 23 , wherein the cell rolling-inducing agent is a selectin or a CTC binding fragment of a selectin. 
     
     
         25 . The device of  claim 24 , wherein the selectin is selected from the group consisting of E-selectin, P-selectin, and L-selectin. 
     
     
         26 . The device of  claim 23 , wherein the capturing agent is adapted to specifically bind to a moiety on a CTC cell surface, and the capture agent is selected from the group consisting of an antibody, an antibody fragment, an engineered antibody, folic acid, transferrin, a peptide, and an aptamer. 
     
     
         27 . The device of  claim 26 , wherein the antibody is anti-EpCAM. 
     
     
         28 . The device of  claim 26  wherein the peptide is an RGD peptide. 
     
     
         29 . The device of  claim 23 , wherein the capturing agent is immobilized on the cell capture surface by direct attachment of the capturing agent to the cell capture surface. 
     
     
         30 - 56 . (canceled)