Sample preparation devices, kits and methods

Abstract

The present disclosure pertains to sample preparation devices useful for affinity capture and purification that include one or more internal structures that comprise a reservoir, a well, a fluid passageway, sorbent particles, and a filter element that blocks passage of the affinity sorbent particles, which sample preparation devices combine the attributes of both dispersive and flow through designs into a single sample preparation device. The present disclosure also pertains to kits that contain and methods that use such sample preparation devices.

1 . A sample preparation device comprising:
 an internal structure that comprise a reservoir, a well, a fluid passageway, sorbent particles, and a filter element that blocks passage of the affinity sorbent particles;   the reservoir having a first volume, a first end and a second end, the first end having a first opening for receiving fluids and the second end having a second opening that opens into the well;   the well having a second volume, a length, a first end closest to the reservoir, and a second end closest to the fluid passageway, the well having a first internal width at its first end that is greater than a second internal width at its second end;   the fluid passageway having a first end closest to the well and a second end comprising an outlet;   the filter element positioned at the first end of the fluid passageway; and   the affinity sorbent particles forming a sorbent bed within the well, the sorbent bed being positioned adjacent to the filter element and at least partially filling the well;   wherein there is no filter element positioned between the reservoir and the well that blocks passage of the affinity sorbent particles.   
     
     
         2 . The sample preparation device of  claim 1 , (a) wherein the first opening for receiving fluids is capped, (b) wherein the second end of the fluid passageway is capped, or (c) wherein the first opening for receiving fluids is capped and the second end of the fluid passageway is capped. 
     
     
         3 . The sample preparation device of  claim 1 , wherein the well is a tapered well that has a single taper angle, multiple taper angles with discrete transitions therebetween, or a continuous change in taper angle. 
     
     
         4 . The sample preparation device of  claim 1 , wherein the well is a tapered well that comprises a taper angle that ranges from 10 to 70 degrees relative to a longitudinal axis of the well. 
     
     
         5 . The sample preparation device of  claim 1 , wherein the well has a hollow partial conical shape or a hollow partial pyramidal shape. 
     
     
         6 . The sample preparation device of  claim 1 , wherein the first internal width of the well ranges from 2 to 30 times the second internal width of the well. 
     
     
         7 . The sample preparation device of  claim 1 , wherein the first internal width of the well ranges from 2 to 30 mm and the second internal width of the well ranges from 1 to 5 mm. 
     
     
         8 . The sample preparation device of  claim 1 , wherein the well is configured such that the sorbent bed can be suspended in a fluid using a technique selected from shaking on a shaker device, vortex mixing, and aspirating and dispensing using a pipette. 
     
     
         9 . The sample preparation device of  claim 1 , wherein the length of the well ranges from 2 to 20 times the second internal width. 
     
     
         10 . The sample preparation device of  claim 1 , wherein the filter element is selected from a frit and a membrane filter, wherein the filter element is a hydrophobic filter element, or both. 
     
     
         11 . The sample preparation device of  claim 1 , wherein the affinity sorbent particles are selected from crosslinked agarose particles, silica-based particles and polymer-based particles. 
     
     
         12 . The sample preparation device of  claim 1 , wherein surfaces of the affinity sorbent particles include bound affinity ligand. 
     
     
         13 . The sample preparation device of  claim 12 , wherein the affinity ligand is selected from proteins, antibodies, aptamers, affimers, inorganic metal complexes, small organic compounds, hydrophobic molecules, polymer imprinted material, oligonucleotides, and oligosaccharides. 
     
     
         14 . The sample preparation device of  claim 1 , wherein the device comprises a plurality of the internal structures. 
     
     
         15 . The sample preparation device of  claim 14 , wherein the plurality of the internal structures form a multi-well strip, a multi-well plate, or a rack of individual pipette tips. 
     
     
         16 . A kit comprising:
 a device comprising an internal structure that comprises a reservoir, a well, a fluid passageway, sorbent particles, and a filter element that blocks passage of the affinity sorbent particles; the reservoir having a first volume, a first end and a second end, the first end having a first opening for receiving fluids and the second end having a second opening that opens into the well; the well having a second volume, a length, a first end closest to the reservoir, and a second end closest to the fluid passageway, the well having a first internal width at its first end that is greater than a second internal width at its second end; the fluid passageway having a first end closest to the well and a second end comprising an outlet; the filter element positioned at the first end of the fluid passageway; and the affinity sorbent particles forming a sorbent bed within the well, the sorbent bed being positioned adjacent to the filter element and at least partially filling the well; wherein there is no filter element positioned between the reservoir and the well that blocks passage of the affinity sorbent particles and   one or more kit components selected from the following: (a) a buffer solution, (b) a washing solution, (c) an elution solution, (d) a sample diluent solution, (e) an antibody standard, (f) an enzyme, (g) a surfactant, (h) a denaturant, (j) a chaoptrope, (k) a reducing agent, (l) an alkylation reagent, (m) a labeling reagent, (n) an affinity ligand, (o) a sample tray and (p) a collection tray.   
     
     
         17 . A method of performing an affinity isolation and purification procedure, the method comprising:
 adding a sample fluid comprising a target analyte to a reservoir of a device that comprises an internal structure that comprises the reservoir, a well, a fluid passageway, sorbent particles, and a filter element that blocks passage of the affinity sorbent particles; the reservoir having a first volume, a first end and a second end, the first end having a first opening for receiving fluids and the second end having a second opening that opens into the well; the well having a second volume, a length, a first end closest to the reservoir, and a second end closest to the fluid passageway, the well having a first internal width at its first end that is greater than a second internal width at its second end; the fluid passageway having a first end closest to the well and a second end comprising an outlet; the filter element positioned at the first end of the fluid passageway; and the affinity sorbent particles forming a sorbent bed within the well, the sorbent bed being positioned adjacent to the filter element and at least partially filling the well; wherein there is no filter element positioned between the reservoir and the well that blocks passage of the affinity sorbent particles   dispersing the affinity sorbent particles in the sample fluid for a time sufficient for the target analyte to bind to the affinity sorbent particles;   removing the sample fluid from the device through the second end of the fluid passageway, forming a settled bed of loaded affinity sorbent particles having bound target analyte;   adding a washing solution to the reservoir, the washing solution causing unbound substances to be removed from the loaded affinity sorbent particles in the settled bed, without removing the bound target analyte;   removing the washing fluid from the device through the second end of the fluid passageway, thereby removing unbound substances from the settled bed;   adding an elution fluid to the reservoir, the elution fluid causing the target analyte to become unbound from the loaded affinity sorbent particles in the settled bed; and   removing the elution fluid from the device through the second end of the fluid passageway, thereby removing purified target analyte from the settled bed.   
     
     
         18 . The method of  claim 17 , wherein the dispersing is performed by shaking the device on a shaker device, by using a vortex mixer, or by repeatedly aspirating and dispensing the sample fluid and the affinity sorbent particles. 
     
     
         19 . The method of  claim 17 , wherein the sample fluid, the washing fluid, and the elution fluid are removed from the device by centrifugation, by applying a vacuum to the second end of the fluid passageway, by applying a positive pressure to the first opening of the reservoir, or a combination thereof. 
     
     
         20 . The method of  claim 17 , further comprising adding a conditioning buffer to the reservoir of the device and removing the conditioning buffer from the device through the second end of the fluid passageway prior to adding the sample fluid.