Composition, apparatus, and method for separating an analyte from a sample

Abstract

An analyte is separated from a fluid sample by introducing the sample into a cartridge having a sample port and a first flow path extending from the sample port. The first flow path includes an extraction chamber containing a solid support for capturing the analyte from the sample. The cartridge has a second flow path for eluting the captured analyte from the extraction chamber, the second flow diverging from the first flow path after passing through the extraction chamber. The sample is forced to flow through the extraction chamber and into a waste chamber, thereby capturing the analyte with the solid support as the sample flows through the extraction chamber. The captured analyte is then eluted from the extraction chamber by forcing an elution fluid to flow through the extraction chamber and along the second flow path.

1 .- 53 . (canceled) 
     
     
         54 . A cartridge comprising:
 a body having a flow path for a fluid sample, the body comprising:
 (i) an inlet port; 
 (ii) an outlet port; and 
 (iii) a chamber extending between the inlet port and the outlet port to form a capture region; 
   wherein the chamber comprises a plurality of columns arranged in a pattern for facilitating interaction of the fluid sample with the surface of the columns,   wherein the plurality of columns are integrally formed with a wall of the chamber and extend into the chamber, and   wherein the flow path is configured for cell capture.   
     
     
         55 . The cartridge according to  claim 54 , wherein the columns are arranged in the pattern selected from one or more of the group consisting of:
 (1) an arrangement wherein the columns are arranged in rows that are randomly misaligned;   (2) an arrangement wherein the columns are arranged so that the distance between adjacent columns is in the range from 0.2 to 200 μm;   (3) an arrangement wherein the columns have cross-sectional lengths in the range from 2 to 200 μm;   (4) an arrangement wherein the columns have cross-sectional widths in the range of 0.2 to 20 μm;   (5) an arrangement wherein the columns have a ratio of cross-sectional length to cross-sectional width of at least 2:1;   (6) an arrangement wherein the columns have a ratio of cross-sectional length to cross-sectional width of at least 4:1;   (7) an arrangement wherein the columns have a ratio height to width or diameter of at least 2:1;   (8) an arrangement an arrangement wherein the columns have a ratio height to width or diameter of at least 4:1;   (9) an arrangement comprising at least 100 columns; and   (10) an arrangement comprising between 1,000 to 10,000 columns.   
     
     
         56 . The cartridge according to  claim 54 , wherein the number of plurality of columns arranged in the chamber is selected from the group consisting of at least 100 columns and between 1,000 to 10,000 columns. 
     
     
         57 . The cartridge according to  claim 54 , wherein the columns have cross-sectional lengths in the range from 2 to 200 μm. 
     
     
         58 . The cartridge according to  claim 54 , wherein the columns have cross-sectional widths in the range of 0.2 to 20 μm. 
     
     
         59 . The cartridge according to  claim 54 , wherein the ratio of cross-sectional length of the columns to the cross-sectional width of the columns is selected from the group consisting of at least 2:1 and at least 4:1. 
     
     
         60 . The cartridge according to  claim 54 , wherein the cross-sectional sizes of the columns is in the range of about 50 μm to about 100 μm. 
     
     
         61 . The cartridge according to  claim 54 , wherein the plurality of columns is arranged in rows that are randomly misaligned. 
     
     
         62 . The cartridge according to  claim 54 , wherein the distance between adjacent columns is in the range from 0.2 to 200 μm. 
     
     
         63 . The cartridge according to  claim 54 , wherein the flow path provided by the plurality of columns provides for an asymmetrically split flow pattern of the fluid sample. 
     
     
         64 . The cartridge according to  claim 54 , further comprising a flow controller for controlling the flow of the fluid sample. 
     
     
         65 . The cartridge according to  claim 64 , wherein the flow path supports a fluid flow rate of the fluid sample flowing through the chamber and supported by the flow controller is in the range of 0.1 μl/sec to 50 μl/sec. 
     
     
         66 . The cartridge according to  claim 64 , wherein the flow controller is selected from the group consisting of a valve, a flow diverter, a fluid diode, and a pump. 
     
     
         67 . The cartridge according to  claim 54 , wherein the surface of the chamber is coated with a hydrophilic material. 
     
     
         68 . The cartridge according to  claim 54 , wherein the surface of the chamber is coated with a binding moiety having binding affinity to a target cell in the fluid sample. 
     
     
         69 . The cartridge according to  claim 68 , wherein the binding moiety is selected from the group consisting of an antibody, a ligand, a nucleic acid, a polypeptide, a protein and a polysaccharide. 
     
     
         70 . The cartridge according to  claim 68 , wherein the binding moiety is coupled to the surface of the chamber by a linker. 
     
     
         71 . The cartridge according to  claim 68 , wherein when the target cell contacts the binding moiety, the target cell is captured by the binding moiety. 
     
     
         72 . The cartridge according to  claim 71 , further comprising a storage chamber for storing an elution fluid for releasing the captured target cell. 
     
     
         73 . The cartridge according to  claim 72 , further comprising a chamber for receiving the released target cell. 
     
     
         74 . The cartridge according to  claim 54 , wherein the columns have a height equal to the depth of the capture chamber. 
     
     
         75 . The cartridge according to  claim 74 , wherein the depth is at least 100 μm. 
     
     
         76 . The cartridge according to  claim 54 , wherein the vertical dimension of the chamber is in the range of 100 to 5,000 μm and the width of the chamber is in the range of 100 to 3,000 μm. 
     
     
         77 . The cartridge according to  claim 54 , wherein the columns have a hexagonal shape. 
     
     
         78 . The cartridge according to  claim 54 , wherein the cartridge is connected to a handheld or desktop external instrument. 
     
     
         79 . The cartridge according to  claim 78 , wherein the cartridge is connected to the external instrument by means of a card-like section of the cartridge and a mating connector within the external instrument. 
     
     
         80 . A method for capturing a target cell in a fluid sample, the method comprising:
 causing a fluid sample containing the target cell to flow through the chamber of the cartridge according to  claim 54 ,   wherein the surface of the chamber is coated with a binding moiety having binding affinity to the target cell and capable of binding the target cell, and wherein when the target cell contacts the binding moiety, the target cell is captured by the binding moiety.   
     
     
         81 . The method according to  claim 80 , further comprising eluting the target cell. 
     
     
         82 . The method according to  claim 80 , further comprising detecting the target cell.