Ex vivo multi-dimensional system for the separation and isolation of cells, vesicles, nanoparticles and biomarkers

Abstract

Devices and techniques are described that involve a combination of multidimensional electrokinetic, dielectrophoretic, electrophoretic and fluidic forces and effects for separating cells, nanovesicles, nanoparticulates and biomarkers (DNA, RNA, antibodies, proteins) in high conductance (ionic) strength biological samples and buffers. In disclosed embodiments, a combination of continuous and/or pulsed dielectrophoretic (DEP) forces, continuous and/or pulsed field DC electrophoretic forces, microelectrophoresis and controlled fluidics are utilized with arrays of electrodes. In particular, the use of chambered DEP devices and of a properly scaled relatively larger electrode array devices that combines fluid, electrophoretic and DEP forces enables both larger and/or clinically relevant volumes of blood, serum, plasma or other samples to be more directly, rapidly and efficiently analyzed. The invention enables the creation of “seamless” sample-to-answer diagnostic systems and devices. The devices and techniques described can also carry out the assisted self-assembly of molecules, polymers, nanocomponents and mesoscale entities into three dimensional higher order structures.

1 . A sample processing device for separation of biological material in a sample fluid, the device comprising:
 at least one inlet that receives a flow of the sample fluid and directs the sample fluid flow to an electrode array; and   at least one outlet that receives the sample fluid flow from the electrode array;   wherein the electrode array comprises a plurality of electrodes configured such that at least one subsection of the electrodes can be charged differently from remaining subsections of the electrodes and thereby establish differently charged dielectrophoresis (DEP) force regions of the electrodes in accordance with the differently charged electrodes and initiate separation of the biological material according to the differently charged DEP force regions.   
     
     
         2 . A sample processing device according to  claim 1 , wherein the electrode array includes:
 a plurality of alternating current (AC) electrodes that are distributed in a planar region of the device between the inlet and the outlet;   a plurality of direct current (DC) electrodes that are arranged adjacent to the AC electrodes.   
     
     
         3 . A sample processing device according to  claim 2 , wherein the DC electrodes extend outside the planar region of the AC electrodes. 
     
     
         4 . A sample processing device according to  claim 2 , wherein at least one subsection of the AC electrodes receives an AC current having a different current frequency from that of the remaining AC electrodes. 
     
     
         5 . A sample processing device according to  claim 1 , wherein the electrodes are constructed of robust materials resistant to corrosion from electrolysis effects. 
     
     
         6 . A sample processing device according to  claim 1 , wherein an overlay of porous material is located on the electrodes such that the porous material is resistant to corrosion from electrolysis effects. 
     
     
         7 . A sample processing device according to  claim 1 , further including at least one analyte outlet through which a flow of analyte is directed out of the device. 
     
     
         8 . A system for separation of biological material, the system comprising:
 a sample processing device according to  claim 1 ; and   a controller configured to selectively energize the electrodes of the sample processing device and initiate separation of the biological material.   
     
     
         9 . A system according to  claim 8 , further comprising an associated detection system for monitoring the separation process and analysis of a separated biological component of the biological material. 
     
     
         10 . A method of separating biological material, the method comprising:
 receiving the biological material into a sample processing device having an electrode array having a plurality of electrodes configured such that at least one subsection of the electrodes can be charged differently from remaining subsections of the electrodes and thereby establish differently charged dielectrophoresis (DEP) force regions of the electrodes in accordance with the differently charged electrodes;   selectively energizing the electrodes and establishing the differently charged DEP force regions of the electrodes in accordance with the differently charged electrodes; and   separating the biological material according to the differently charged DEP force regions into at least one separated analyte component that is received at an analyte outlet of the sample processing device, and remaining components of the biological material.   
     
     
         11 . A method according to  claim 10 , further including monitoring a separation process and analysis of separated biological components. 
     
     
         12 . A method according to  claim 10 , wherein separating comprises holding at least one type of biological material at one of the electrode subsections, the method further including:
 introducing a reagent into the sample processing device; and   reacting the introduced reagent with the held type of biological material in the sample processing device.   
     
     
         13 . A method according to  claim 12 , wherein the reagent comprises a fluorescent die. 
     
     
         14 . A method according to  claim 12 , wherein the reagent comprises antibodies. 
     
     
         15 . A method according to  claim 12 , wherein reacting comprises performing PCR operations on the held type of biological material. 
     
     
         16 . A system for separation of biological materials, the system comprising:
 two or more separate electrode chamber structures each with and electrode, said groups of electrode chamber structures separated by an inner chamber structure with pores (portals, holes), which allows an electric field to pass from outer electrode chambers through the inner chamber, an inlet into the inner chamber that receives the biological materials, at least one analyte outlet that receives a separated component of the biological materials from the inner chamber, and another outlet that receives remaining components of the biological materials;   a controller configured to selectively energize the electrodes and initiate separation of the biological material and associated detection system for monitoring separation process and analysis of separated biological components.   
     
     
         17 . A method of separating biological materials, the method comprising:
 receiving the biological materials into an inner chamber structure with outer electrode chamber structures, an inlet that receives the biological materials, at least one analyte outlet that receives a separated component of the biological materials from the inner chamber, an outlet that receives remaining components of the biological materials, and an associated detection system for monitoring separation process and analysis of separated biological components; selectively energizing the electrodes; and   separating the biological material into at least one separated analyte component that is received at an analyte outlet of the inner chamber structure, and remaining components of the biological material.   
     
     
         18 . A method where the systems of any of  claims 11  through  15  are used to carry out the assisted self-assembly of molecules, polymers, nanocomponents, including DNA and protein derivatized nanoparticles, quantum dots, nanotubes, and the like, and mesoscale components into higher order three dimensional structures, materials, and devices.