METHODS AND SYSTEMS FOR DRUG DISCOVERY AND SUSCEPTIBILITY ASSAY IN USING A FERROFLUID

Abstract

A system for determining drug effectiveness on a plurality of cells is described. The system includes flowing a ferrofluid mixed with a plurality of biological cells through an inlet portion of a cartridge, the cartridge comprising a plurality of microfluidic channels, the inlet is in communication with a portion of each of the plurality of channels, applying a magnetic field proximate at least one of the inlet portion and the plurality of micro-channels, wherein the magnetic field is configured to apply an indirect force on the mix, separating biologic cells according to at least a first type as the mix flows in a first direction; and directing at least the first type of cells toward a first sensor functionalized with receptors via at least one of the micro-channels, the sensor arranged proximate to a second portion of at least one of the micro-channels downstream from the first inlet portion.

Claims

1-22. (canceled)

23. A system for determining drug effectiveness on a plurality of cells comprising: a cartridge comprising a plurality of micro-channels; an inlet receiving a ferrofluid and biological cell mixture, the inlet in communication with at least one of the plurality of micro-channels; a magnetic field source arranged proximate to at least one of the inlet and the plurality of micro-channels; at least one receptor region having receptors for binding with at least a first type of biological cell, the at least one receptor region arranged proximate to a first portion of at least one of the micro-channels, wherein the first portion is downstream from the inlet; at least one quartz-crystal-microbalance (QCM) sensor arranged proximate to the first portion of at least one of the micro-channels; and a controller; wherein: the magnetic field source generates a magnetic field that applies a force on the mixture so as to separate at least biological cells of the first type from the mixture, at least a first micro-channel of the plurality of micro-channels receives biological cells of the first type and directs the first type of cells to the at least one receptor region, each of the plurality of micro-channels receives a dosage of a drug to interact with the first type of biological cell captured by the receptors, and the controller has computer instructions operating thereon to cause the controller to determine, using the at least one QCM sensor: a first mass corresponding to the first type of cells captured by the receptors before the drug is received, and a second mass corresponding to the first type of cells after the drug interacts with the first type of cells captured by the receptors.

24. The system of claim 23, wherein separating at least the biological cells of the first type from the mix comprises at least one of separating, focusing and concentrating.

25. The system of claim 23, wherein the at least one receptor region comprises a plurality of receptor regions, each receptor region being functionalized with a specific receptor for at least one particular type of biological cell and each receptor corresponding to a specific micro-channel of the plurality of micro-channels; and the magnetic field applies a force on the biological cells in the mix to separate a plurality of types of biological cells from the mix and direct the types of cells into one and/or another micro-channel.

26. The system of claim 23, wherein the first type of biological cell comprises a biological cell of a predetermined size, shape, weight, charge and/or configuration.

27. The system of claim 23, further comprising thermal managing means surrounding at least one of the cartridge, the first micro-channel, and the remainder of the micro-channels to substantially maintain the micro-channels at a first temperature.

28. The system of claim 23, wherein the drug is received by at least one of: the cartridge, the inlet, and a second inlet into one or more of the plurality of micro-channels.

29. The system of claim 23, wherein the controller has computer instructions operating thereon to cause the controller to determine, based on the first mass and the second mass, a susceptibility of the first type of cells to the first drug.

30. The system of claim 29, wherein the controller has computer instructions operating thereon to determine, based on the first mass and the second mass, the cell growth rate of the first type of cells.

31. The system of claim 30, wherein the controller has operating thereon computer instructions to cause the controller to track a signal of the at least one QCM sensor, such that an increase in mass corresponds to an increase in the total cell volume of the first type of cells and tracks changes in the total mass of the cells bound to the surface.

32. The system of claim 23, wherein the controller has computer instructions operating thereon to cause the controller to determine: a first cell growth rate of the captured first type of cells before the drug is received, and a second cell growth rate of the captured first type of cells after the drug interacts with the captured first type of cells.

33. The system of claim 32, wherein the controller has computer instructions operating thereon to cause the controller to determine susceptibility of the first type of cells to the drug by comparing the first cell growth rate and the second cell growth rate.

Description

BRIEF DESCRIPTION OF THE DRAWING

[0022] FIG. 1 is a schematic of a drug susceptibility assay system, according to some embodiments of the present disclosure.

[0023] FIG. 2 is an enlarged schematic of a drug susceptibility assay system according to some embodiments of the present disclosure.

DETAILED DESCRIPTION OF SOME OF THE EMBODIMENTS

[0024] FIG. 1 illustrates a drug susceptibility assay system, utilizing a ferrofluid, according to some embodiments. As shown, an initial sample containing a mixture of cells 2 is mixed with a ferrofluid 3 (e.g., a biocompatible ferrofluid) in a reservoir 1. An external force, such as a pressure source, e.g., a pump 4, introduces the overall mixture into a channel inlet 8 that is connected to fluidic channel 5 that sits atop a magnetic field source 6. The magnetic field source 6, which is configured to apply a force, either directly or indirectly to particles/cells 2 of the mix (e.g., on non-magnetic particles/cells), such that the cells 2 are forced upward and focused. The magnetic source 6 may comprise at least one of a planar electrode(s), an electromagnet(s) and a permanent magnet(s), each of which may be arranged in an array. In some embodiments, the cells move along the channel ceiling (e.g., roll) and interact serially with receptor regions 7 on that surface. Specific interactions between the cells 2 and the receptor regions 7 result in the temporary, and in some embodiments permanent, capture of cells. In some embodiments, the captured cells are allowed to seed and grow over a predetermined period of time, for establishing their viability.

[0025] Thereafter, according to some embodiments, a predetermined dose of a drug is introduced into the flow, and the growth rate of the cells (or their morbidity) in response to the drug exposure is determined by a detecting means. In some embodiments, detecting means, such as an optical scanner 10, may be provided and configured to detect the target particles captured and/or moving along the receptor regions 7, such detecting means may also comprise, in addition or in place of, one or more of, for example: U.S. Pat. No. 4,448,534, WO2013/155525, WO2008/042003, U.S. Pat. No. 8,364,409, WO1991/001381, WO2013/054311 (as well as other detecting means familiar to those of skill in the art. The optical scanner may be, in some embodiments, impedance sensors, quartz crystal microbalance (QCM) sensors or surface plasmon resonance (SPR) sensors. The mixture flows through to the channel outlet 9, in some embodiments, to waste or back to the reservoir 1.

[0026] FIG. 2 illustrates a drug susceptibility assay utilizing a ferrofluid according to some embodiments. An initial sample containing a mixture of cells 23 mixed with ferrofluid 22 is configured to flow through a fluidic channel 21 that sits atop a magnetic field source 26 (for example). The magnetic field source 26, which may be configured to apply a force, either directly or indirectly to particles/cells 23 of the mix (e.g., on non-magnetic particles/cells), such that the cells 23 are forced upward and focused. The magnetic source 26 may comprise at least one of a planar electrode(s), an electromagnet(s) and a permanent magnet(s), each of which may be arranged in an array. In some embodiments, the cells move along the channel ceiling (e.g., roll) and interact serially with receptor regions 24 on that surface. In some embodiments, hydrodynamic barrier 25 is placed between receptor regions. Specific interactions between the cells 23 and the receptor regions 24 result in the temporary, and in some embodiments permanent, capture of cells.

[0027] This invention discloses a system and a method that combines the sensitivity of an impedance sensor or QCM with the sample manipulation, isolation and capture capability of a ferrofluidic device. A complex sample (such as whole blood) is mixed with a biocompatible ferrofluid and is introduced into a disposable cartridge that is placed on top of a magnetic field source (integrated current-carrying electrodes on a printed circuit board or its combination with other magnetic sources). As the ferrofluid-sample mixture is circulated within the fluidic network of the disposable cartridge, rare target cells or pathogens are separated, sorted, extracted, focused and directed towards the sensor surface that is functionalized with antibodies or other receptors corresponding to the specific cell or pathogen of interest. The target moieties are strongly pushed towards the sensor surface and are rapidly captured. The sensor channel is then flushed continuously with culture media and kept at an optimal culture temperature via thermal management hardware surrounding the cartridge.

[0028] Initially, the culture is allowed to grow (for up to 1 hour) to ensure that there are viable organisms captured over the sensor surface. Afterwards, an appropriate dosage of drug is introduced into the media. If the cells change the rate of their growth or stop dividing altogether, the susceptibility to the introduced drug may be quantified from the changes in the growth curve of those cells.

[0029] The signal from the functionalized sensor is taken differentially with respect to a non-functionalized sensor of matching geometry within the same channel. As the cells grow and divide, their total volume increases, leading to an increased differential signal on the impedance sensor. Similarly, if a QCM is used, changes in the total mass of all cells bound to the functionalized surface result in the signal.

[0030] Impedance and QCM based sensor geometries have been used to characterize cell cultures and/or determine drug susceptibility in the past. The advantage of the present invention is that it can extract and capture cells directly from a complex and large-volume sample without any additional sample preparation or pre-culture steps. Hence, within 2 hours of sample collection, drug susceptibility testing may be completed within this platform.

[0031] Another major advantage of this invention is its multiplexing capability. Bioferrofluidic sample extraction and purification is quite rapid, and the purified cells may be directed towards a very small sensor surface with great accuracy within the same cartridge. Hence, dozens of sensor surfaces may be used within a single cartridge to run simultaneous drug susceptibility tests of many different cell species.

[0032] Any and all references to publications or other documents, including but not limited to, patents, patent applications, articles, webpages, books, etc., presented in the present application, are herein incorporated by reference in their entirety.

[0033] Example embodiments of the devices, systems and methods have been described herein. As noted elsewhere, these embodiments have been described for illustrative purposes only and are not limiting. Other embodiments are possible and are covered by the disclosure, which will be apparent from the teachings contained herein. Thus, the breadth and scope of the disclosure should not be limited by any of the above-described embodiments but should be defined only in accordance with claims supported by the present disclosure and their equivalents. Moreover, embodiments of the subject disclosure may include methods, systems and devices which may further include any and all elements from any other disclosed methods, systems, and devices, including any and all elements corresponding to drug discovery and susceptibility. In other words, elements from one or another disclosed embodiments may be interchangeable with elements from other disclosed embodiments. In addition, one or more features/elements of disclosed embodiments may be removed and still result in patentable subject matter (and thus, resulting in yet more embodiments of the subject disclosure). Correspondingly, some embodiments of the present disclosure may be patentably distinct from one and/or another reference by specifically lacking one or more elements/features. In other words, claims to certain embodiments may contain negative limitation to specifically exclude one or more elements/features resulting in embodiments which are patentably distinct from the prior art which include such features/elements.