Methods and devices for high throughput purification

Abstract

Described herein are devices and methods for high throughput purification of particles. In some cases, methods and devices described herein can be used to remove erythrocytes and purify leukocytes and raise the quality of umbilical cord blood and other transplant grafts, thereby significantly improving patient outcomes.

1 . A method for transferring leukocytes in a blood sample to a fluid medium, comprising flowing a blood sample and the fluid medium through separate inlets of a microfluidic device, wherein said device comprises:
 a) at least one microfluidic channel having at least two inlets and a plurality of outlets; and   b) an array of obstacles arranged in rows and columns within the microfluidic channel, wherein each subsequent row of obstacles is shifted laterally with respect to a previous row such that the obstacles differentially deflect cells based on size; and wherein the obstacles comprise a shape that is asymmetric to an axis parallel to a bulk flow direction of the fluid medium;   
       wherein the blood sample is flowed at a flow rate of at least 100 ml/hour per array of obstacles, and wherein flowing the sample through the microfluidic channel transfers leukocytes larger than a predetermined size to the fluid medium. 
     
     
         2 . The method of  claim 1 , wherein the blood sample is undiluted. 
     
     
         3 . The method of  claim 1 , wherein the blood sample is diluted up to five-fold. 
     
     
         4 . The method of  claim 1 , wherein the blood sample is prepared in the form an apheresis or leukapheresis sample. 
     
     
         5 . The method of  claim 1 , wherein the fluid medium is a buffer medium. 
     
     
         6 . The method of  claim 1 , wherein the blood sample is at least 100 mL. 
     
     
         7 . The method of  claim 1 , wherein the concentration of leukocytes in the blood sample is at least 1×10 6  cells/mL. 
     
     
         8 . The method of  claim 1 , wherein the microfluidic device comprises at least five microfluidic channels in parallel. 
     
     
         9 . The method of  claim 1 , wherein each of the obstacles comprises a shape which comprises a cross-section which comprises a triangle, a square, a rectangle, a pentagon, or a hexagon. 
     
     
         10 . The method of  claim 1 , wherein the leukocytes comprise T cells. 
     
     
         11 . The method of  claim 1 , wherein the leukocytes comprise CD34+ cells. 
     
     
         12 . The method of  claim 1 , wherein a ratio of the radius of curvature of an edge to an adjacent side of each of the obstacles is no greater than 0.25. 
     
     
         13 . The method of  claim 1 , wherein each of the obstacles comprises a shape which comprises three or more points. 
     
     
         14 . A microfluidic device for separating particles by size, the microfluidic device comprising:
 a) at least one microfluidic channel having at least two inlets and a plurality of outlets; and   b) an array of obstacles arranged in rows and columns within the microfluidic channel, wherein each subsequent row of obstacles is shifted laterally with respect to a previous row such that the obstacles differentially deflect cells based on size; and wherein the obstacles comprise a shape that is asymmetric to an axis parallel to a bulk flow direction of the fluid medium;   wherein the device is configured such that leukocytes can be transferred to a fluid medium from a blood sample flowed through the device at a flow rate of at least 100 ml/hour per array of obstacles.   
     
     
         15 . The microfluidic device of  claim 14 , wherein each of the obstacles comprises a shape which comprises a cross-section which comprises a triangle, a square, a rectangle, a pentagon, or a hexagon. 
     
     
         16 . The microfluidic device of  claim 15 , wherein the microfluidic device comprises at least five microfluidic channels in parallel. 
     
     
         17 . The microfluidic device of  claim 14 , wherein each of the obstacles comprises a shape which comprises three or more points. 
     
     
         18 . The microfluidic device of  claim 17 , wherein a ratio of the radius of curvature of an edge to an adjacent side of each of the obstacles is no greater than 0.25. 
     
     
         19 . The microfluidic device of  claim 14 , wherein the rows and columns of obstacles are laterally shifted by a periodic tilt angle. 
     
     
         20 . The microfluidic device of  claim 19 , wherein the rows and columns of obstacles form a lattice. 
     
     
         21 . The method of  claim 1 , wherein the microfluidic device comprises at least thirty microfluidic channels. 
     
     
         22 . The method of  claim 1 , wherein the microfluidic device comprises at least one-hundred microfluidic channels. 
     
     
         23 . The microfluidic device of  claim 14 , wherein the device comprises at least thirty microfluidic channels. 
     
     
         24 . The microfluidic device of  claim 14 , wherein the device comprises at least one-hundred microfluidic channels.