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 which comprises
 an inlet port, 
 an outlet port, and 
 a chamber extending between the inlet port and the outlet port to form a capture region, 
   wherein the flow path is designed to support a fluid flow rate of 0.1 μl/sec to 50 μl/sec,   wherein the chamber comprises a plurality of columns arranged in a pattern of differing sizes, shapes, and/or spacing between each other for facilitating the interaction of a fluid sample with the surface of the columns,   wherein the pattern has the characteristics of
 1) the columns having cross-sectional lengths in the range of from 2 to 200 μm and being integrally formed with at least one wall of the chamber and extending into the chamber, 
 2) the columns are arranged so that the spacing between adjacent columns varies, and 
 3) the combination of spacing between the columns and different cross-sectional lengths and different cross-sectional widths of the columns determines the volume of the capture region occupied by the columns, 
   wherein the flow path is configured for capturing cells, and   when in operation, the chamber is connected to a flow controller for controlling the flow of the fluid sample through the cartridge at a flow rate of 0.1 μl/see to 50 μl/sec.   
     
     
         55 . The cartridge according to  claim 54 , wherein the pattern or volume occupied by the columns in the capture region is further determined by a combination of arrangements of the columns selected from the group consisting of:
 (1) arranging the columns in rows that are randomly misaligned,   (2) arranging the columns so that the distance between adjacent columns is in the range from 0.2 to 200 μm,   (3) arranging columns having cross-sectional lengths in the range from 2 to 200 μm,   (4) arranging columns having cross-sectional widths in the range of 0.2 to 20 μm,   (5) arranging columns having a ratio of cross-sectional length to cross-sectional width of at least 2:1,   (6) arranging columns having a ratio of cross-sectional length to cross-sectional width of at least 4:1,   (7) arranging columns having a ratio of height to width or diameter of at least 2:1,   (8) arranging columns having a ratio of height to width or diameter of at least 4:1,   (9) arranging at least 100 columns, and   (10) arranging between 1,000 to 10,000 columns.   
     
     
         56 . The cartridge according to  claim 54 , wherein the columns are aligned substantially perpendicular to the at least one wall of the chamber. 
     
     
         57 . The cartridge according to  claim 54 , wherein the distance between adjacent columns is in the range from 0.2 to 200 μm. 
     
     
         58 . The cartridge according to  claim 54 , wherein the surface of the chamber is coated with a hydrophilic material. 
     
     
         59 . 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. 
     
     
         60 . 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, and 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. 
     
     
         61 . The cartridge according to  claim 54 , wherein a binding moiety is coupled to the surface of the chamber by a linker. 
     
     
         62 . The cartridge according to  claim 54 , wherein the chamber is covered with a top portion. 
     
     
         63 . The cartridge according to  claim 54 , further comprising a separate inlet port for introducing a fluid. 
     
     
         64 . The cartridge according to  claim 54 , wherein the columns are arranged in a pattern of rows that are randomly misaligned. 
     
     
         65 . A method for capturing a target cell in a sample 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 is capable of binding the target cell.   
     
     
         66 . The method according to  claim 65 , further comprising detecting the target cell. 
     
     
         67 . The method according to  claim 65 , wherein the fluid sample flows through the chamber at a speed of 0.1 μl/sec to 50 μl/sec. 
     
     
         68 . A cartridge comprising:
 a body having a flow path for a fluid sample, the body comprising:
 an inlet port, 
 an outlet port, and 
 a chamber extending between the inlet port and the outlet port to form a capture region, 
   wherein the flow path is designed to support a fluid flow rate of the fluid sample of 0.1 μl/sec to 50 μl/sec,   wherein the chamber comprises a plurality of columns arranged in a pattern of differing sizes, shapes, and/or spacing between each other for facilitating the interaction of the fluid sample with the surface of the columns,   wherein the pattern is designed according to the following parameters
 1) the columns having cross-sectional lengths in the range of from 2 to 200 μm and being integrally formed with at least one wall of the chamber and extending into the chamber, 
 2) the columns are arranged so that the spacing between adjacent columns varies, and 
   3) the combination of spacing between the columns and different cross-sectional lengths and different cross-sectional widths of the columns determines the volume of the capture region occupied by the columns,   wherein the flow path is configured for capturing cells, and   when in operation, the chamber is connected to a flow controller for controlling the flow of the fluid sample through the cartridge at a flow rate of 0.1 μl/sec to 50 μl/sec.   
     
     
         69 . The cartridge according to  claim 68 , wherein the pattern or volume occupied by the columns in the capture region is further determined by a combination of arrangements of the columns selected from the group consisting of:
 (1) arranging the columns in rows that are randomly misaligned,   (2) arranging the columns so that the distance between adjacent columns is in the range from 0.2 to 200 μm,   (3) arranging columns having cross-sectional lengths in the range from 2 to 200 μm,   (4) arranging columns having cross-sectional widths in the range of 0.2 to 20 μm,   (5) arranging columns having a ratio of cross-sectional length to cross-sectional width of at least 2:1,   (6) arranging columns having a ratio of cross-sectional length to cross-sectional width of at least 4:1,   (7) arranging columns having a ratio of height to width or diameter of at least 2:1,   (8) arranging columns having a ratio of height to width or diameter of at least 4:1,   (9) arranging at least 100 columns, and   (10) arranging between 1,000 to 10,000 columns.   
     
     
         70 . The cartridge according to  claim 68 , wherein the columns are arranged in a pattern of rows that are randomly misaligned. 
     
     
         71 . A cartridge comprising:
 a body having a flow path which comprises
 an inlet port, 
 an outlet port, and 
 a chamber having a capture region extending between the inlet port and the outlet port, 
   wherein the capture region comprises a plurality of upstanding columns substantially perpendicular to at least one wall of the chamber and aligned transverse to the flow path and arranged in a pattern of differing sizes, shapes, and/or spacing between each other for facilitating the interaction of a fluid sample with the surface of the columns such that there can be no straight-line flow through the capture region and that streamlined flow streams are disrupted, the columns being integrally formed with at least one wall of the chamber and extending into the chamber,   wherein the combination of spacing between columns, different cross-sectional lengths of the columns and different cross-sectional widths of the columns determines the volume of the capture region occupied by the columns.   
     
     
         72 . The cartridge according to  claim 71 , wherein the volume of the columns with respect to the volume of the capture region is characterized by a combination of arrangements of the columns selected from the group consisting of:
 (1) arranging the columns in rows that are randomly misaligned,   (2) arranging the columns so that the distance between adjacent columns is in the range from 0.2 to 200 μm,   (3) arranging columns having cross-sectional lengths in the range from 2 to 200 μm,   (4) arranging columns having cross-sectional widths in the range of 0.2 to 20 μm,   (5) arranging columns having a ratio of cross-sectional length to cross-sectional width of at least 2:1,   (6) arranging columns having a ratio of cross-sectional length to cross-sectional width of at least 4:1,   (7) arranging columns having a ratio of height to width or diameter of at least 2:1,   (8) arranging columns having a ratio of height to width or diameter of at least 4:1,   (9) arranging at least 100 columns, and   (10) arranging between 1,000 to 10,000 columns.   
     
     
         73 . The cartridge according to  claim 71 , wherein the surface of the chamber is coated with a binding moiety having binding affinity to a target cell in the fluid sample. 
     
     
         74 . The cartridge according to  claim 71 , wherein the surface of the chamber is coated with a binding moiety having binding affinity to a target cell in the fluid sample, and 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. 
     
     
         75 . The cartridge according to  claim 71 , wherein a binding moiety is coupled to the surface of the chamber by a linker. 
     
     
         76 . The cartridge according to  claim 71 , further comprising a separate inlet port for introducing a fluid. 
     
     
         77 . The cartridge according to  claim 71 , wherein the columns are arranged in a pattern of rows that are randomly misaligned.