Systems and methods for particle identification, selection, and purification - Patents.com

Abstract

A method for purifying particles includes generating charged particles from a sample, measuring at least one of the mass, charge magnitude, and mobility of the generated charged particles, and selectively passing each measured charged particle having at least one of (a) a measured mass equal to or within a selected particle mass range, (b) a measured charge magnitude equal to or within a selected charge magnitude range, (c) a mass-to-charge ratio equal to or within a selected mass-to-charge ratio range, and (d) a measured mobility equal to or within a selected mobility range to a particle collection target. In some embodiments, the collected particles can be harvested and amplified.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This patent application claims the benefit of and priority to U.S. Provisional Patent Application Serial No. 62 / 913460, filed October 10, 2019, U.S. Provisional Patent Application Serial No. 62 / 949559, filed December 18, 2019, and U.S. Provisional Patent Application Serial No. 62 / 972403, filed February 10, 2020, the disclosures of which are expressly incorporated herein by reference in their entireties.

[0002] Government Rights

[0002] This invention was made with Government support under Grant GM131100 awarded by the National Institutes of Health. The United States Government has certain rights in this invention.

[0003] Technical Field

[0003] The present disclosure relates generally to devices and methods for identifying, selecting, and purifying particles, and more specifically to devices and methods for identifying, selecting, and purifying particles based on one or more molecular characteristics. [Background technology]

[0004]

[0004] Spectrometry instruments provide identification of the chemical composition of a substance by measuring one or more molecular properties of the substance. Some such instruments are configured to analyze substances in solution, while others are configured to analyze charged particles of substances in the gas phase. The molecular information produced by many such charged particle measurement instruments is limited in the number and types of molecular properties that can be measured. Thus, purification using such instruments is similarly limited. Summary of the Invention

[0005]

[0005] The present disclosure may include one or more of the features recited in the appended claims and / or one or more of the following features and combinations thereof: In one configuration, a particle purification device may include an ion generator configured to generate charged particles from a sample, an ion processing region configured to receive the charged particles generated by the ion generator and measure at least one of a mass and a charge magnitude of the generated charged particles, a particle collection target, means for selectively passing charged particles exiting the ion processing region to the particle collection target, a processor, and a memory storing instructions executable by the processor for controlling the means for selectively passing charged particles to pass each of the measured charged particles having at least one of (a) a measured mass equal to or within a selected particle mass range, (b) a measured charge magnitude equal to or within a selected charge magnitude range, and (c) a mass-to-charge ratio equal to or within a selected mass-to-charge ratio range to the particle collection target.

[0006]

[0006] In another configuration, a method for purifying particles may include generating charged particles from a sample, measuring at least one of the mass and charge magnitude of the generated charged particles, and selectively passing each of the measured charged particles having at least one of (a) a measured mass equal to or within a selected range of particle masses, (b) a measured charge magnitude equal to or within a selected range of charge magnitudes, and (c) a mass-to-charge ratio equal to or within a selected range of mass-to-charge ratios to a particle collection target.

[0007]

[0007] In yet another configuration, a method for purifying particles may include generating charged particles from a sample, measuring charge magnitudes of the generated charged particles, and selectively passing each of the measured charged particles having a measured charge magnitude equal to or within a selected charge magnitude range to a particle collection target.

[0008] In yet another aspect, a method for purifying particles includes generating charged particles from a sample; measuring the mass of the generated charged particles; and selectively passing each of the measured charged particles having a measured mass equal to or within a range of the selected mass to a particle collection target.

[0009]

[0009] In a further configuration, a method for purifying particles may include generating charged particles from a sample, measuring the mass and charge magnitude of the generated charged particles, calculating the mass-to-charge ratio of the measured charged particles based on the measured mass and charge magnitude, and selectively passing each of the measured charged particles having a calculated mass-to-charge ratio equal to or within a selected mass-to-charge ratio range to a particle collection target.

[0010]

[0010] In another further configuration, a method for purifying particles may include generating charged particles from a sample, measuring at least one of the mass, charge magnitude, and mobility of the generated charged particles, and selectively passing each of the measured charged particles having at least one of (a) a measured mass equal to or within a selected particle mass range, (b) a measured charge magnitude equal to or within a selected charge magnitude range, (c) a mass-to-charge ratio equal to or within a selected mass-to-charge ratio range, and (d) a measured mobility equal to or within a selected mobility range to a particle collection target.

[0011] In another further aspect, a method for purifying particles includes generating charged particles from a sample; measuring the mobility of the generated charged particles; and selectively passing each of the measured charged particles having a measured mobility equal to or within a range of the selected mobility to a particle collection target.

[0012]

[0012] In another further configuration, a method for measuring particles in a sample of extracellular vesicles may include generating ions from the sample of extracellular vesicles and measuring the mass and charge of at least a subset of the generated ions using a charge detection mass spectrometer.

[0013]

[0013] In yet another configuration, a method for measuring exosomes in a sample aliquot may include generating ions from the sample aliquot, measuring the mass and charge of at least some of the generated ions using a charge detection mass spectrometer, and identifying a subset of the measured ions that are exosome ions from the measured masses of at least some of the generated ions. [Brief description of the drawings]

[0014] [Figure 1] FIG. 1 is a simplified diagram of an apparatus and process for purifying particles. [Diagram 2] FIG. 2 is a simplified flowchart of an embodiment of a process for controlling the apparatus of FIG. 1 to generate and measure charged particles and to generate a resultant spectrum from which a subpopulation of charged particles can be identified or selected for purification. [Diagram 3] FIG. 3 is a simplified flow chart of an embodiment of a process for controlling the apparatus of FIG. 1 to purify particles by generating, measuring, and filtering charged particles, and for collecting the purified particles. [Figure 4A] FIG. 4A is a scatter plot of particle charge versus mass produced from a urinary exome sample by an embodiment of the apparatus of FIG. 1, where the ion processing region is implemented in the form of a charge detection mass spectrometer. [Figure 4B] FIG. 4B superimposes on the plot of FIG. 4A an exemplary selection of subpopulations of particles by the apparatus of FIG. 1 for purification, where the selected subpopulations are defined by ranges of particular particle mass values. [Figure 4C] FIG. 4C superimposes on the plot of FIG. 4A another exemplary selection of particle subpopulations for purification, where the selected subpopulations are defined by a range of particular particle charge values. [Figure 4D] FIG. 4D superimposes on the plot of FIG. 4A yet another example selection of particle subpopulations for purification, where the selected subpopulations are defined by a particular range of particle charge values ​​and a particular range of particle mass values. [Figure 4E] FIG. 4E superimposes on the plot of FIG. 4A yet another exemplary selection of particle subpopulations for purification, where the selected subpopulations are defined by ranges of mass-to-charge ratio values ​​for particular particles. [Figure 4F]FIG. 4F overlays the plot of FIG. 4A with a further example selection of particle subpopulations for purification, where the selected subpopulations are defined by a range of specific particle mass-to-charge ratio values ​​and a range of specific particle mass values. [Figure 4G] FIG. 4G overlays on the plot of FIG. 4A yet another example selection of particle subpopulations for purification, where the selected subpopulations are defined by a range of specific particle mass-to-charge ratio values ​​and a range of specific particle charge values. [Figure 4H] FIG. 4H superimposes on the plot of FIG. 4A yet another exemplary selection of particle subpopulations for purification, where the selected subpopulations are defined by a range of specific particle mass-to-charge ratio values, a range of specific particle mass values, and a range of specific particle charge values. [Diagram 5] FIG. 5 is a simplified flow chart of another process embodiment for controlling the apparatus of FIG. 1 to identify, collect, and / or purify populations and / or subpopulations of specified types of charged particles. [Figure 6A] FIG. 6A is a scatter plot of charge versus mass of particles produced from an exosome-enriched bovine milk sample by an embodiment of the apparatus of FIG. 1 and using the process illustrated in FIG. 5, where the ion processing region of the apparatus is implemented in the form of a charge detection mass analyzer. [Figure 6B] FIG. 6B overlays multiple boundaries onto the scatter plot of FIG. 6A, illustrating the processing of the plotted data into various subpopulations of charged particles. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0015]

[0028] For the purposes of promoting an understanding of the principles of the present disclosure, reference will now be made to a number of exemplary embodiments illustrated in the accompanying drawings, and specific language will be used to describe the same.

[0016]

[0029] This disclosure relates to devices and techniques for identifying and / or purifying particles based on one or more molecular properties, examples of which may include, but are not limited to, mass, charge, mass-to-charge ratio, mobility, etc. For purposes of this disclosure, the terms "charged particle" and "ion" may be used interchangeably, and both terms are intended to refer to any particle having a net positive or negative charge. The terms "purify" and "purification" are intended to refer to the identification and extraction, i.e., separation, of a subpopulation of charged particles generated from a sample based on one or more molecular properties.

[0017]

[0030] Referring now to FIG. 1, a diagram of an apparatus 10 for purifying particles is shown. FIG. 1 further illustrates an exemplary process 12 for collecting the purified particles and, in some embodiments, processing the collected and purified particles. In the illustrated embodiment, the apparatus 10 includes, by way of example, an ion source region 14 having an outlet coupled to an inlet of a charged particle processing region 16. The outlet of the charged particle processing region 16 is coupled to an inlet of a charged particle deflector (CPD) or steering device (CPSD) 18. In some embodiments, the apparatus 10 further includes, optionally, a conventional ion trap (IT) 20 including an inlet and an outlet opposite the inlet, as illustrated by the dashed lines in FIG. 1, which is coupled to the outlet of the charged particle deflector or steering device 18. In such an embodiment, the outlet of the ion trap 20 defines the charged particle outlet of the apparatus 10. In other embodiments in which the ion trap 20 is omitted, the outlet of the apparatus 10 is the outlet of a charged particle deflector or steering device 18 .

[0018]

[0031] The ion source region 14 illustratively includes an ion generator 22 configured to generate ions, i.e., charged particles, from a sample 24. The ion generator 22 illustratively is implemented in the form of any conventional device or apparatus for generating ions from a sample. As one example, which should not be considered limiting in any way, the ion generator 22 can be or include a conventional electrospray ionization (EIS) source, a matrix-assisted laser desorption ionization (MALDI) source, or other conventional ion generator configured to generate ions from the sample 24. The sample 24 for generating ions can be any biological or other material. In some embodiments, the sample 24 is dissolved, dispersed, or carried in a solution, while in other embodiments, the sample may not be in or part of a solution.

[0019]

[0032] In an exemplary embodiment, the voltage source VS1 is electrically connected to the processor 26 via a number of signal paths F, which may be any positive integer, and further electrically connected to the ion source region 14 via a number of signal paths G, which may also be any positive integer. In some embodiments, the voltage source VS1 may be implemented in the form of a single voltage source, while in other embodiments, the voltage source VS1 may include any number of separate voltage sources. In some embodiments, the voltage source VS1 may be configured or controlled to create and provide one or more time-invariant (i.e., DC) voltages of selectable magnitude. Alternatively or additionally, the voltage source VS1 may be configured or controlled to create and provide one or more switchable time-invariant voltages, i.e., one or more switchable DC voltages. Alternatively or additionally, the voltage source VS1 may be configured or controllable to create and provide one or more time-varying signals of selectable shape, duty cycle, peak magnitude, and / or frequency.

[0020]

[0033] The processor 26 may, by way of example, be conventional and may include a processing circuit or multiple processing circuits. The processor 26 may, by way of example, include or be coupled to a memory 28 that stores instructions that, when executed by the processor 26, cause the processor 26 to control the voltage source VS1 to produce one or more output voltages for selectively controlling the operation of the ion generator 22. In some embodiments, the processor 26 may be implemented in the form of one or more conventional microprocessors or controllers, and in such embodiments, the memory 28 may be implemented in the form of one or more conventional memory units that store instructions in the form of one or more instructions or instruction sets executable by a microprocessor. In other embodiments, the processor 26 may alternatively or additionally be implemented in the form of a field programmable gate array (FPGA) or similar circuit, and in such embodiments, the memory 28 may be implemented in the form of a programmable logic block included in the FPGA and / or external to the FPGA, within which instructions may be programmed and stored. In yet another embodiment, processor 26 and / or memory 28 may be implemented in the form of one or more application specific integrated circuits (ASICs). Those skilled in the art will recognize other ways in which processor 26 and / or memory 28 may be implemented, and it will be understood that any such other ways of implementation are anticipated by and are considered to be within the scope of this disclosure. In some alternative embodiments, voltage source VS1 may itself be programmable to selectively produce one or more constant and / or time-varying output voltages.

[0021]

[0034] In the illustrated embodiment, the voltage source VS1 is configured to generate one or more voltages that cause the ion generator 22 to generate ions from the sample 24 in response to a control signal generated by the processor 26, for example. In some embodiments, the sample 24 is disposed within the ion source region 14, as illustrated in FIG. 1, while in other embodiments, the sample 24 can be disposed outside the ion source region 14. In one illustrated embodiment, which should not be considered limiting in any way, the sample 24 is provided in the form of a solution and the ion generator 22 is a conventional electrospray ion (EIS) source that is configured to generate ions from the sample 24 in the form of a fine mist of charged droplets in response to one or more voltages provided by VS1. As mentioned above, it will be understood that ESI and MALDI represent only two examples of many conventional ion generators, and that the ion generator 22 can be or include any such conventional device or apparatus for generating ions from a sample, whether in the form of a solution or not.

[0022]

[0035] The ion processing region 16 includes, by way of example, a number M of ion processing stages or devices 161-16, which may be any positive integer. M one or more ion processing devices 161-16 M may, for example, be operable to process the charged particles generated in the ion source region 14 and transmitted to the ion processing region 16, such as by measuring one or more molecular properties of the charged particles, by filtering the charged particles based on the one or more molecular properties to provide a subpopulation or subset of charged particles having at least one specified molecular property, and / or by separating, e.g., breaking down, the charged particles.

[0023]

[0036] In an exemplary embodiment, the voltage source VS2 is electrically connected to the processor 26 via a number H of signal paths, which may be any positive integer, and further electrically connected to the ion processing region 16 via a number J of signal paths, which may also be any positive integer. In some embodiments, the voltage source VS2 may be implemented in the form of a single voltage source, while in other embodiments, the voltage source VS2 may include any number of individual voltage sources. In some embodiments, the voltage source VS2 may be configured or controlled to create and provide one or more time-invariant (i.e., DC) voltages of selectable magnitude. Alternatively or additionally, the voltage source VS2 may be configured or controlled to create and provide one or more switchable time-invariant voltages, i.e., one or more switchable DC voltages. Alternatively or additionally, the voltage source VS2 may be configured or controllable to create and provide one or more time-varying signals of selectable shape, duty cycle, peak magnitude, and / or frequency. Generally, one or more outputs of voltage source VS2 may be coupled to, for example, one or more ion processing devices 161-16 in ion processing region 16. M and the number of such outputs and / or the type (one or more) of voltages produced thereat are determined by one or more of the ion processing devices 161-16. M It will be appreciated that the number and / or type of ion processing device(s) forming the ion processing region 16 may depend on the number and / or type of ion processing device(s) forming the ion processing region 16. In any case, the memory 28 may illustratively have instructions stored therein that, when executed by the processor 26, cause the processor 26 to control the one or more ion processing devices 161-16 of the ion processing region 16. M The inverter is adapted to control a voltage source VS2 to produce one or more output voltages for selectively controlling the operation of the inverter.

[0024]

[0037] Ion Processing Device (1 or more) 161-16 MExamples of the ion processing device(s) 161-16 may include, but are not limited to, one or more devices and / or apparatus for separating, collecting, and / or filtering the charged particles according to one or more molecular properties, in any order and / or combination. Examples of the one or more devices and / or apparatus for separating the charged particles according to one or more molecular properties may include, but are not limited to, one or more mass spectrometers or mass analyzers, one or more ion mobility spectrometers, one or more gas chromatographs, etc. Ion processing device(s) 161-16 may include, but are not limited to, one or more mass spectrometers. M Examples of mass analyzers in this embodiment include, but are not limited to, any mass analyzer operative to at least measure the mass-to-charge ratio of ions and to direct the measured ions from the mass analyzer to the charged particle deflector or steering device 18. In such embodiments where the mass analyzer is operative to only measure the mass-to-charge ratio of ions, the mass analyzer may be conventional. In other such embodiments, the mass analyzer may be provided, for example, in the form of a mass analyzer configured to measure both the mass and charge magnitudes of the charged particles generated in the ion source region 14 and directed to the ion processing region 16. In one example of this embodiment, which should not be considered limiting in any way, the mass analyzer may be implemented, for example, in the form of a charge-detecting mass analyzer (CDMS), in which case the ion processing device(s) 161-16 may be coupled to the ion processing device(s) 161-16. MThe ion processing device(s) 161-162 of FIG. 1 includes a conventional through-ion mass analyzer or mass analyzer and one or more corresponding CDMS charge detectors. In some embodiments, the one or more CDMS charge detectors can be provided in the form of one or more electrostatic linear ion traps (ELITs), and in other embodiments, the one or more CDMS charge detectors can be provided in the form of at least one orbitrap. In some embodiments, the CDMS detector(s) can include at least one ELIT and at least one orbitrap. Although CDMS is, by way of example, a single-particle technique that typically operates to measure the mass and charge magnitude values ​​of a single ion, some CDMS detectors are designed and / or operated to measure the mass and charge of more than one charged particle at a time. The ion processing device(s) 161-162 of FIG. M Some examples of CDMS devices and / or techniques that may be implemented in or as part of a mass spectrometer, and some examples of CDMS charge detectors and / or techniques, are disclosed in co-pending International Patent Application Nos. PCT / US2019 / 013251, PCT / US2019 / 013274, PCT / US2019 / 013277, PCT / US2019 / 013278, PCT / US2019 / 013280, PCT / US2019 / 013283, PCT / US2019 / 013284, and PCT / US2019 / 013285, all of which were filed on January 11, 2019, the disclosures of which are expressly incorporated herein by reference in their entireties.

[0025]

[0038] In another embodiment including a mass analyzer configured to measure both the mass and charge magnitude of the charged particles generated in the ion source region 14 and transmitted to the ion processing region 16, such a mass analyzer can be provided in the form of a conventional mass analyzer (e.g., a quadrupole mass analyzer, etc.) configured to selectively pass ions of a specified mass-to-charge ratio or ions within a specified range of mass-to-charge ratios, or in the form of a similarly configured through-ion mass analyzer, in either case followed by an electric field-free drift region including a charge detector array (CDA) configured to measure the charge magnitude or charge state of the charged particles emerging from the mass analyzer or mass analyzer. The ion processing device(s) 161-16 in FIG. M Some example configurations of such mass analyzers that may be implemented as or as part of the mass analyzer are disclosed in co-pending U.S. patent application Ser. No. 62 / 949555 and / or co-pending U.S. patent application Ser. No. 62 / 949554, both of which were filed on December 18, 2019, the disclosures of which are expressly incorporated herein by reference in their entireties.

[0026]

[0039] Ion processing device(s) 161-16 as described above M In some embodiments including a mass analyzer configured to measure both the mass and charge of the charged particles provided by the ion source region 14, as shown in Figure 1, the associated charge detector(s) or charge detector array are electrically connected to respective inputs(s) of a number N of charge detection amplifiers CA, the outputs(s) of the number N of charge detection amplifiers CA being electrically connected to a processor 26, where N can be any positive integer. Each of the charge amplifiers CA, by way of example, is conventional and is responsive to charge induced by the charged particles at the one or more individual charge detectors to produce a corresponding charge detection signal at its output and provide the charge detection signal to the processor 26.

[0027]

[0040] In any embodiment that includes one or more conventional mass analyzers, such mass analyzers may be provided in the form of one or any combination of a time-of-flight (TOF) mass analyzer, a reflector mass analyzer, a Fourier transform ion cyclotron resonance (FTICR) mass analyzer, a quadrupole mass analyzer, a triple quadrupole mass analyzer, a magnetic sector mass analyzer, an orbitrap mass analyzer, and the like.

[0028]

[0041] Ion processing device (one or more) including one or more ion mobility spectrometers 161-16 M In embodiments, examples of ion mobility spectrometers include, but are not limited to, single-tube linear ion mobility spectrometers, multiple-tube linear ion mobility spectrometers, circular-tube ion mobility spectrometers, etc. Ion processing device(s) 161-161 may include one or more devices and / or apparatus for collecting charged particles. M In the embodiment, examples of the one or more devices and / or apparatus for collecting charged particles include, but are not limited to, a quadrupole ion trap, a hexapole ion trap, an ion funnel, etc. Ion processing device(s) 161-16 including one or more devices and / or apparatus for filtering charged particles. M In the embodiment, examples of the one or more devices and / or apparatus for filtering charged particles include, but are not limited to, one or more devices or apparatus for filtering charged particles according to mass-to-charge ratio, one or more devices or apparatus for filtering charged particles according to particle mobility, etc. Ion processing device(s) 161-16 including one or more devices and / or apparatus for separating charged particles. MIn embodiments, examples of the one or more devices and / or apparatus for separating charged particles include, but are not limited to, one or more devices or apparatus for separating charged particles by collision-induced dissociation (CID), surface-induced dissociation (SID), electron capture dissociation (ECD), and / or photoinduced dissociation (PID), etc.

[0029]

[0042] Ion Processing Device (1 or more) 161-16 M It will be understood that the ion processing device(s) 161-16 may include one or any of the above apparatus, devices, or stages in any order and in any combination, and some embodiments may include multiple adjacent or separate ones of any such apparatus, devices, or stages. M The ion processing device(s) 161-16 may include one CDMS configured to measure the mass and charge of the charged particles as described above and then provide the measured charged particles to the charged particle deflector or steering device 18. As another non-limiting implementation of the apparatus 10 illustrated in FIG. M As briefly described above, the ion processing device(s) 161-16 may include a mass analyzer including a charge detector array configured to measure the mass and charge of the charged particles and to provide the measured charged particles to the charged particle deflector or steering device 18. In any of these examples, the processor 26 is illustratively programmed to control the voltage source VS2 to cause the mass analyzer to measure the mass and charge of the charged particles. As yet another non-limiting implementation of the apparatus 10 illustrated in FIG. 1, the ion processing device(s) 161-16 may include a mass analyzer including a charge detector array configured to measure the mass and charge of the charged particles and to provide the measured charged particles to the charged particle deflector or steering device 18. Mmay include a mass-to-charge ratio filter, for example in the form of a quadrupole mass analyzer. In this example, the processor 26 is illustratively programmed to control a voltage source VS2 to cause the mass-to-charge ratio filter to selectively pass only ions having a specified mass-to-charge ratio, or ions having a mass-to-charge ratio within a specified range of mass-to-charge ratios, towards the charged particle deflector or steering device 18. In some such embodiments, the ion processing device(s) 161-16 M may further include a mass analyzer disposed between the mass-to-charge ratio filter and the charged particle deflector or steering device 18, which is configured to measure the mass and charge of the charged particles exiting the mass-to-charge ratio filter. In some such embodiments, the ion processing device(s) 161-16 M may further include a particle separation stage or device disposed between the mass-to-charge ratio filter and the mass analyzer, which is configured to separate the charged particles exiting the mass-to-charge ratio filter. In such an embodiment, the processor 26 is illustratively programmed to control the voltage source VS2 to thereby operate the illustrative device(s) and / or stage(s) in a conventional manner. M It will be understood that other examples and combinations of the above will occur to those of ordinary skill in the art, and such examples and combinations are considered to be within the scope of this disclosure. In any case, the processor 26 controls the voltage source VS2 to thereby power the ion processing device(s) 161-16. M to operate in a conventional manner and / or as described herein.

[0030]

[0043] In embodiments including this, the charged particle deflector or charged particle steering device 18 is configured, for example, to selectively pass only those charged particles having one or more specified molecular properties or having one or more molecular properties within a range of molecular properties through its outlet, while the remaining charged particles are blocked, in the case of a charged particle deflector, for example by steering such charged particles towards a conductive structure, and in the case of a charged particle steering device, are directed away from the outlet where the charged particles are collected, for example through another passageway or outlet where the charged particles are not collected or stored.

[0031]

[0044] In one exemplary embodiment, the charged particle deflector or steering device 18 can be implemented in the form of a conventional single inlet, single outlet charge detector configured and controllable to selectively pass or block ions. In another exemplary embodiment, the charged particle deflector or steering device 18 can be implemented in the form of a conventional single inlet, multiple outlet charge steering device configured and controllable to selectively steer ions entering the single inlet through one of a number of different ion outlets for collecting purified charged particles. In either case, another voltage source VS3 is electrically connected to the processor 26 via a number K of signal paths, which may be any positive integer, and further electrically connected to the charged particle deflector or steering device 18 via a number L of signal paths, which may also be any positive integer. In some embodiments, the voltage source VS3 can be implemented in the form of a single voltage source, while in other embodiments the voltage source VS3 can include any number of individual voltage sources. In some embodiments, voltage source VS3 can be configured or controlled to generate and provide one or more time-invariant (i.e., DC) voltages of selectable magnitude. Alternatively or additionally, voltage source VS3 can be configured or controlled to generate and provide one or more switchable time-invariant voltages, i.e., one or more switchable DC voltages. Alternatively or additionally, voltage source VS3 can be configured or controllable to generate and provide one or more time-varying signals of selectable shape, duty cycle, peak magnitude, and / or frequency. It will be understood that in general, one or more outputs of voltage source VS3 are coupled to, by way of example, charged particle deflector or steering device 18, and that the number of such outputs and / or the type (s) of voltages generated thereat are dependent on the type of charged particle deflector or steering device 18 implemented.In either case, the memory 28 illustratively has instructions stored therein that, when executed by the processor 26, cause the processor 26 to control the voltage source VS3 to produce one or more output voltages for selectively controlling the operation of the charged particle deflector or steering device 18.

[0032]

[0045] In some embodiments where the charged particle deflector or steering device 18 is implemented in the form of a single inlet, single outlet charge detector, the processor 26 is illustratively operative to deflect the charged particles entering the inlet towards a conductive structure, e.g., a conductive plate, tube, or rod, by controlling the voltage source VS3 to create an electric field E large enough to drive and accelerate the charged particles P towards the conductive structure. The processor 26 is illustratively operative in such embodiments to direct the charged particles entering the inlet through the outlet, by controlling the voltage source VS3 to create conditions, e.g., small or no electric field, within the deflector that allow the charged particles to pass therethrough. In some embodiments where the charged particle deflector or steering device 18 is implemented in the form of a single inlet, multiple outlet charge detector, the processor 26 is operative, for example, to steer ions entering the inlet into a path where the purified charged particles are not collected and / or through an outlet by controlling the voltage source VS3 to create an electric field E of sufficient magnitude to steer the charged particles P through such an outlet. The processor 26 is operative, in such embodiments, for example, to steer the charged particles entering the inlet through an outlet where the purified charged particles are collected by controlling the voltage source VS3 to create conditions in the charge steering device that allow the charged particles to pass through an individual outlet. Many alternative embodiments of the charged particle deflector or steering device 18 are illustrated and described in U.S. Patent Application Serial No. 62 / 52 / 949555, filed December 18, 2019, which is incorporated herein by reference, although it will be understood that such embodiments are provided by way of example only. Other charged particle deflection and / or steering apparatus or devices will occur to those skilled in the art, and it will be understood that any other such charged particle deflection and / or steering apparatus or devices are considered to be within the scope of this disclosure.

[0033]

[0046] In some embodiments, the ion trap 20 may be coupled to the charged particle deflector or steering device 18, as briefly described above and illustrated in dashed line representation in FIG. 1. In such embodiments, a further voltage source VS4 is electrically connected to the processor 26 via a number P of signal paths, which may be any positive integer, and further electrically connected to the ion trap 20 via a number Q of signal paths, which may also be any positive integer. In some embodiments, the voltage source VS4 may be implemented in the form of a single voltage source, while in other embodiments, the voltage source VS4 may include any number of individual voltage sources. In some embodiments, the voltage source VS4 may be configured or controlled to create and provide one or more time-invariant (i.e., DC) voltages of selectable magnitudes. Alternatively or additionally, the voltage source VS4 may be configured or controlled to create and provide one or more switchable time-invariant voltages, i.e., one or more switchable DC voltages. Alternatively or additionally, voltage source VS4 can be configured or controllable to produce and provide one or more time-varying signals of selectable shape, duty cycle, peak magnitude, and / or frequency. One or more outputs of voltage source VS4 are illustratively coupled to ion trap 20, and memory 28 illustratively stores therein instructions that, when executed by processor 26, cause processor 26 to control ion trap 20 to produce one or more output voltages for controlling ion trap 20 to selectively trap and store charged particles within ion trap 20, and to selectively release and accelerate trapped particles from ion trap 20.

[0034]

[0047] Processor 26 is further coupled to one or more peripheral devices (PD) 30 via a number R of signal paths, which may be, by way of example, any positive integer. In some embodiments, peripheral device 30 includes at least one of a conventional display monitor, a printer, and / or other output device, and in such embodiments, memory 28 stores instructions that, when executed by processor 26, cause processor 26 to control one or more such output peripheral devices 30 to display and / or record an analysis of the operation of apparatus 10, including, by way of example, but not limited to, particle spectrum information measured by apparatus 10.

[0035]

[0048] As briefly described above, by way of example, the apparatus 10, under the control of the processor 26 via the control of the voltage sources VS1, VS2, VS3, and in some embodiments VS4, is operable to purify the charged particles generated by the ion generator 22 by selectively passing only a subpopulation or subset of the generated charged particles having one or more molecular properties or having molecular properties within a range of one or more molecular properties. In such an embodiment, for example, the subpopulation or subset may include, by way of example, only charged particles having a specified mass or masses within a specified mass range. In another embodiment, the subpopulation or subset may include, by way of example, only charged particles having a specified charge or masses within a specified charge magnitude or charge state range. In yet another embodiment, the subpopulation or subset may include, by way of example, only charged particles that are of a specified charge or charge magnitude or charge state range and are a specified mass, or only charged particles that are of a specified charge or charge magnitude or charge state range and are a specified mass range. In yet another embodiment, the subpopulation or subset may, for example, include only charged particles of a specified mass-to-charge ratio or only charged particles having a mass-to-charge ratio value within a specified range of mass-to-charge ratio values. In some such embodiments, the subpopulation or subset may further include only such charged particles that also have a specified mass value or have a mass value within a specified range of mass values, and / or only charged particles that also have a specified charge magnitude or charge state value or have a charge magnitude or charge state value within a specified range of charge magnitude or charge state values. In yet another embodiment, the subpopulation or subset may, for example, include only charged particles of a specified mobility or only charged particles having a mobility within a specified range of mobility values.In some such embodiments, the subpopulations or subsets may be further limited to a specified charged particle mass value or range of mass values, or to a specified charge magnitude or charge state or a specified range, or to a specified mass-to-charge ratio or a specified range of charged particle devices 161-16 implemented in any particular embodiment of device 10. M The number and / or type (one or more) of charged particle devices 161-16 may also be dependent on the particular subpopulation or subset of charged particles desired to be purified, and the number and / or type (one or more) of charged particle devices 161-16 may be determined by ... M Those of skill in the art will recognize that a variety of different types and combinations of can be used. Additionally, those of skill in the art will be aware of other molecular feature subpopulations or subsets and / or combinations thereof that may be sought for purification, and it will be understood that such other molecular feature subpopulations or subsets and / or combinations thereof, and various devices and device combinations for collecting them, are considered to be within the scope of this disclosure.

[0036]

[0049] Also depicted in Figure 1 is a simplified process 12 for collecting particles and, in some embodiments, processing the collected and purified particles. In some embodiments, for example, a subpopulation or subset of the charged particles exiting the apparatus 10 is collected at a surface 40A of a particle collection target 40 via particle deposition, e.g., low energy deposition, or via other conventional particle collection techniques. The particle collection target 40, or at least its surface 40A, is illustratively a non-reactive or inert material that does not bind or react with the purified charged particles exiting the apparatus 10. In some embodiments, the particle collection surface 40A of the particle collection target 40 can be viscous or oily, or otherwise configured or constructed therein such that the purified charged particles exiting the apparatus 10 can be effectively collected over a period of time. In another embodiment in which an ion trap 20 is included, the purified charged particles exiting the device 10 may be trapped and collected in the ion trap 20 for a period of time and then released en masse from the ion trap 20 to a particle collection surface 40A of a particle collection target 40. In either case, the particle collection surface 40A of the particle collection target 40 is illustratively configured to not only collect the purified charged particles exiting the device 10, but also to provide for collection of the collected and purified particles therefrom. In some embodiments, for example, the purified particles collected at the particle collection surface 40A of the particle collection target 40 may be collected by rinsing the surface 40A with a solution 45 provided from a solution source 42 and directing the resulting combination 46 of the purified particle-bearing solution 45 to a suitable container 44. It will be understood that those skilled in the art will recognize other techniques, apparatus, devices, etc. for harvesting the purified charged particles collected at surface 40A of particle collection target 40, and any other such techniques, apparatus, devices, etc. are contemplated as being within the scope of this disclosure.

[0037]

[0050] In some embodiments, the harvested purified particle population can be amplified, i.e., replicated or multiplied, with a conventional particle amplifier or particle amplification process 48. In implementations in which the purified particles are or include DNA, for example, the particle amplifier or amplification process 48 can be in the form of, for example, a conventional polymerase chain reaction (PCR) device or process for amplifying or replicating the particles over several orders of magnitude, e.g., thousands or millions of copies. It will be understood that one of skill in the art will recognize other devices and / or processes for amplifying harvested purified particles, whether or not the harvested purified particles are or include DNA, and any such other particle amplification devices and / or processes are considered to be within the scope of this disclosure.

[0038]

[0051] In some cases, it may be desirable to view all or at least a portion of the molecular property spectrum of the sample 24 to identify or facilitate identification of subpopulations or subsets thereof for purification. In this regard, FIG. 2 shows a simplified flow chart illustrating a process 100 for operating the apparatus 10 of FIG. 1 to measure one or more molecular properties of charged particles generated from the sample 24 and to process such measurements to produce a multi-dimensional, e.g., two or more dimensional, molecular property spectrum. At least some steps of the process 100 are stored in the memory 28 in the form of instructions executable by the processor 26 to measure, analyze, and visualize the spectrum. The process 100 begins at step 102, where the processor 26 illustratively operates to control the voltage source VS1 to cause the ion generator 22 to generate charged particles from the sample 24 and transmit the generated charged particles to the ion processing region 16. Thereafter, in step 104, the processor 26 operates to control the voltage source VS2, thereby causing one or more instruments or devices in the ion processing region 16 to measure two or more molecular properties.

[0039]

[0052] In some embodiments, the ion processing region 16 may include or be implemented in the form of a mass analyzer configured to measure the mass of the particles and the charge of the particles, as described above with respect to FIG. 1. In some such embodiments, for example, such a mass analyzer may be implemented in the form of a charge detection mass analyzer (CDMS), while in other embodiments, such a mass analyzer may be implemented in the form of a mass analyzer followed by a charge detection array (CDA), a mass-to-charge filter, or other device configured to measure mass-to-charge ratio (conventional MS), some examples of which are illustrated and described in U.S. Patent Application Serial No. 62 / 949555, filed December 18, 2019, which is incorporated herein by reference. In other embodiments, the ion processing region 16 may include or be implemented in the form of an ion mobility spectrometer (IMS) followed by such a charge detection array. In yet other embodiments, the ion processing region 16 may include or be implemented in the form of a combination of mass analyzers, ion mobility spectrometers, and charged particle charge measurement devices or devices. In some such embodiments, for example, the ion processing region 16 may include an IMS followed by a CDMS, or an IMS followed by a conventional MS followed by a CDA. In other such embodiments, by way of further example, the ion processing region 16 may include a conventional MS followed by an IMS followed by a CDA, or a conventional MS followed by a CDA followed by an IMS, or a CDMS followed by an IMS. In these exemplary embodiments of the ion processing region 16, the processor 26, by way of example, operates to control the voltage source VS2 in step 104, such that the spectrometer device(s) measures the charge magnitude or charge state of the generated charged particles and the mass and / or mobility values ​​of the generated charged particles, as illustrated in FIG. 2.

[0040]

[0053] Following step 104, processor 26 is operative to process the measurements made in step 104 and generate a charged particle spectrum therefrom. In one example where sample 24 is a solution of urinary exosomes and ion processing region 16 is implemented in the form of a conventional MS or CDMS followed by a CDA, processor 26 is illustratively operative in step 106 to generate a scatter plot of the magnitude of the charge of the charged particle (in units of elementary charge, e) versus the mass of the charged particle (in units of megadaltons, MDa), as shown in FIG. 4A.

[0041]

[0054] Following step 106, process 100 proceeds to step 108, where the spectrum produced in step 106 is analyzed, e.g., visually or automatically, by processor 26 to determine an appropriate subpopulation or subset of particles for purification. The subpopulation or subset of particles may be selected based on one or any combination of the following values ​​or ranges (one or more) of the particle's mass, mass-to-charge ratio, charge (size or charge state), or mobility, by way of example.

[0042]

[0055] 3, there is shown a simplified flow chart of a process 200 for purifying particles from a sample 24 using any of the various embodiments of the apparatus 10 shown in FIG. 1. In some embodiments, the implementation of the apparatus 10 used to perform the process 100 shown in FIG. 2 can also be used to perform the process 200 shown in FIG. 3 following the process 100. In other embodiments, for example, where the values ​​and / or ranges of molecular properties for purification are known in advance, the process 100 illustrated in FIG. 2 can be avoided and the configuration of the apparatus 10 can be specifically selected to achieve or facilitate the desired purification. In any case, at least some steps of the process 200 can be stored in the memory 28, for example, in the form of instructions executable by the processor 26 to perform the purification of a selected subpopulation or subset of charged particles generated from the sample 24 illustrated in FIG. 1. Process 200 begins at step 202, where processor 26 illustratively operates to control voltage source VS1 to cause ion generator 22 to generate charged particles from sample 24 and provide the generated charged particles to ion processing region 16. Thereafter, at step 204, processor 26 controls voltage source VS2 to cause one or more apparatus or devices in ion processing region 16 to measure two or more molecular properties. Various combinations of apparatus or devices can be implemented in ion processing region 16 to measure any two or more molecular properties, and some examples of such apparatus or devices and such one or more molecular properties are provided in the description of process 100 above.In these exemplary embodiments of the ion processing region 16, the processor 26 is, by way of example, operative to control the voltage source VS2 in step 204, which causes the spectrometer device(s) to measure the charge magnitude or charge state of the generated charged particles and the mass and / or mobility values ​​of the generated charged particles, as shown in the example of FIG. 3, although it will be understood that the ion processing region 16 in step 204 may alternatively be implemented in a different manner, i.e., with different devices, and / or the one or more molecular properties may be measurable molecular properties other than or in addition to the particle's mass, mass-to-charge ratio, mobility, and charge (magnitude or charge state).

[0043]

[0056] Following step 204, the process 200 proceeds to step 206, where the processor 26 operates to control the voltage source VS3 such that the charged particle deflector or steering device 18 causes only charged particles within a selected subpopulation or subset of the charged particle beam generated by the ion generator 22 to pass through its charged particle outlet or through a designated one of its multiple charged particle outlets. As discussed above, the subpopulation or subset of charged particles generated by the ion generator 22 may be selected based on one or any combination of measurements of the particle's mass, mass-to-charge ratio, charge (magnitude or charge state), or mobility value(s) or range(s). Once such measurements are known to processor 26 as individual charged particles exit ion processing region 16, processor 26 operates to control charged particle deflector or steering device 18 via control of voltage source VS3 to selectively pass for collection only those charged particles having one or a combination of measured molecular characteristic values ​​as defined by a selected subpopulation or subset of the charged particles generated by ion generator 22. For purposes of illustrating the operation of steps 204 and 206 of process 200, several example subpopulations or subsets of the urinary exosome spectrum illustrated in FIG. 4A, and several example configurations and implementations of apparatus 10 for purifying such subpopulations, are described below with respect to FIGS. 4B-4H.

[0044]

[0057] Following step 206, process 200 proceeds to step 208, where charged particles exiting charged particle deflector or charged particle steering device 18 through its one outlet or through a selected one of its multiple outlets are collected. In an embodiment of apparatus 10 including ion trap 20, step 208 illustratively includes control of voltage source VS4 by processor 26 to supply one or more voltages to ion trap 20 such that ion trap 20 collects and stores such charged particles exiting charged particle deflector or charged particle steering device 18 through its one outlet or through a selected one of its multiple outlets. Following expiration of the collection time period during which the ion trap 20 operates to collect and store the exiting charged particles therein, the processor 26 is further operative to control the voltage source VS4 to provide one or more voltages to the ion trap 20, in step 208, thereby causing the ion trap 20 to release the stored ions and direct them towards the collection surface 40A of the collection target 40. In an embodiment of the apparatus 10 that does not include an ion trap 20, step 208 illustratively includes collecting the charged particle discharge onto the collection surface 40A of the collection target 40 as the charged particles exit the charged particle deflector or charged particle steering device 18. Thereafter, in step 210, the purified charged particles collected on the collection surface 40A of the collection target 40 are collected, for example, as described above with respect to FIG. 1. In some embodiments, the process 200 includes another step 212 following step 210, in which amplification, i.e., replication or multiplication, of the collected particles is performed in a conventional manner as described above.

[0045]

[0058] 5, there is shown a simplified flow chart of a process 500 for controlling any of the various embodiments of the apparatus 10 shown in FIG. 1 to identify, collect, and / or purify a population and / or subpopulation of charged particles of a specified type to purify particles from a sample 24. The process 500 begins at step 502, where a sample is provided in which particles of a specified type are present. The specified particles can be any particle, e.g., a molecule, or collection thereof, that is within or part of a cell, and / or that is transported between cells, and that has a mass in the megadalton range or greater. Examples of the types of particles present in the sample, and for which the sample is selected and provided, can be or include, but are not limited to, exosomes, endosomes, microvesicles in general, ectosomes, apoptotic bodies, retroviruses, exomeres, chylomicrons, DNA, RNA, proteins, lipids, acids, carbohydrates, enzymes, viruses, bacteria, and the like. Other examples of samples and / or particles of interest present in a sample that are considered to be within the scope of this disclosure include, but are not limited to, any cell that releases exosomes or extracellular vesicles, that is or is part of any biological tissue(s), fluid(s), and / or other biological material(s), any molecule or collection of molecules that is encapsulated in a bio-layer, such as, for example, a virus, any non-compartmentalized organelle that is grouped but not bound by or within a bio-layer, any extracellular vesicle that has been modified, for example, by the addition of one or more small molecules or by the addition of a drug, such as an anti-cancer drug, to produce a detectable mass shift.

[0046]

[0059] In some embodiments, the sample provided in step 502 in which the particular type of particles is present may be sample 24 shown in FIG. 1 from which charged particles are generated for analysis in apparatus 10. In some alternative embodiments, process 500 may include step 504, shown in dashed outline, in which the sample provided in step 502 is enriched for the particular particle type. Following step 504, in embodiments that include it, process 500 may, by way of example and in one embodiment, proceed to step 506, in which the enriched sample 24, i.e., the sample provided in step 502 and enriched for the particular particle type in step 504, is used to perform process 100 as illustrated in FIG. 2. In embodiments that do not include step 504, step 502 is followed by step 506, in which case process 100 as illustrated in FIG. 2 is performed using sample 24 in which the specified type of particles is present. In some embodiments, step 108 of process 100 in which subpopulations of particle spectra are identified and / or selected for refinement can include performing one or more conventional statistical and / or modeling processes on a data set of particles to identify and / or select one or more subpopulations of particles. An example of such a statistical process is described below with respect to Example 8.

[0047]

[0060] In some embodiments, process 500 ends after performing step 506. In some alternative embodiments, process 500 proceeds from step 506 to step 508, where the enriched sample 24 is used to perform process 200 as illustrated in Figure 3 to purify the specified type of particles or one or more subpopulations thereof identified in step 506. In some alternative embodiments, as described above with respect to Figure 3, process 500 can proceed to step 508, directly from step 504 in embodiments that include step 504, or directly from step 502 in embodiments that do not include step 504.

[0048]

[0061] In some embodiments including step 504, the process(es) used to enrich the sample for the specified particle type may depend on the sample type and / or the specified particle type, and in any case will be known to those of skill in the art. In such embodiments, the enriched sample resulting from step 504 is sample 24, shown in FIG. 1, from which charged particles are generated for analysis by device 10. An example of such a process used to enrich exosomes from a bovine milk sample is described below in Example 8, but should not be considered limiting in any way. In other embodiments, including or not including step 504, various configurations and / or implementations of ion processing region 16 of device 10 may be used to enrich and / or assist in enriching the sample for the specified particle type. For example, some embodiments may include unwanted particles known to be present in one or more ranges of particle mass, mass-to-charge ratio, and / or mobility that differ from the range(s) of particle mass, mass-to-charge ratio, and / or mobility in a sample of a specified type, and in such embodiments, ion processing region 16 may be variously configured to filter out some or all of such unwanted particles prior to performing steps 506 and / or 508, as described above.

[0049] example Example 1

[0062] 4B, the urinary exome plot of FIG. 4A is reproduced with an example of the selection of a subpopulation or subset 300 of particles by the apparatus of FIG. 1 for purification according to steps 204 and 206 of the process 200 illustrated in FIG. 3. In this example, the selected subpopulation 300 is simply defined by a specified range of particle mass values, between 20 MDa and 30 MDa. In order for the processor 26 to control the voltage source VS3 in step 206 to cause the charged particle deflector or charged particle steering device 18 to pass therethrough only charged particles having particle masses within the specified particle mass range of 20-30 MDa towards the particle target 40, the particle measurement information produced by the one or more apparatus or devices of the ion processing region 16 must include particle mass information or particle measurement information that allows the processor 26 to determine particle mass prior to step 206. 4A, the ion processing region 16 is illustratively implemented in the form of a conventional MS followed by a CDA or a CDMS, either configured to measure particle mass directly or to determine particle mass from measurements of charged particles made by the apparatus(es) in step 204. However, it will be appreciated that the ion processing region 16 may alternatively be or include any apparatus or device, or combination of apparatus or devices, configured to measure particle mass or configured to measure one or more properties or characteristics of a particle from which particle mass can be determined or estimated.In either case, once the particle mass information has been determined in step 204, in step 206 the processor 26 operates to control the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 passes the charged particle exiting the ion processing region 16 towards the particle target 40 only if the mass of the particle is within a specified particle mass range of 20-30 MDa, otherwise controls the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 either blocks the passage of the particle towards the target 40 or steers the charged particle away from the target 40.

[0050] Example 2

[0063] 4C, the urinary exome plot of FIG. 4A is reproduced with another example of the selection of another subpopulation or subset 302 of particles by the apparatus of FIG. 1 for purification according to steps 204 and 206 of the process 200 illustrated in FIG. 3. In this example, the selected subpopulation 302 is simply defined by a specified range of particle charge magnitude values ​​between 750e and 900e. In order for the processor 26 to control the voltage source VS3 in step 206 to cause the charged particle deflector or charged particle steering device 18 to pass therethrough only charged particles having particle charge values ​​within the specified particle charge range of 750-900e towards the particle target 40, the particle measurement information produced by the one or more apparatus or devices of the ion processing region 16 prior to step 206 must include particle charge information or particle measurement information that allows the processor 26 to determine particle charge. 4A, ion processing region 16 is illustratively implemented in the form of a conventional MS followed by a CDA or a CDMS, either configured to measure the charge of the particles directly or to determine the charge of the particles from measurements of the charged particles made by an apparatus(es) in step 204. However, it will be appreciated that ion processing region 16 may alternatively be or include any apparatus or device, or combination of apparatus or devices, configured to measure the charge of the particles or configured to measure one or more characteristics or properties of the particles from which the charge of the particles can be determined or estimated.In either case, once the particle charge information has been determined in step 204, in step 206 the processor 26 operates to control the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 passes the charged particle exiting the ion processing region 16 towards the particle target 40 only if the particle's charge is within a specified particle charge magnitude range of 750-900e, otherwise controls the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 blocks the passage of the particle towards the target 40 or steers the charged particle away from the target 40.

[0051] Example 3

[0064] Referring now to Figure 4D, the urinary exome plot of Figure 4A is further reproduced with an example of selection of yet another subpopulation or subset 304 of particles by the apparatus of Figure 1 for purification according to steps 204 and 206 of process 200 illustrated in Figure 3. In this example, the selected subpopulation 304 is defined by a specified range of particle mass values ​​between 10 MDa and 15 MDa, and a specified range of particle charge magnitude values ​​between 600e and 700e. In order for the processor 26 to control the voltage source VS3 in step 206 such that the charged particle deflector or steering device 18 passes only charged particles having a particle mass value within a specified particle mass value range of 10-15 MDa and a charge value within a specified particle charge range of 600-750e towards the particle target 40, the particle measurement information produced by the one or more apparatus or devices in the ion processing region 16 must include particle mass and charge information or particle measurement information that allows the processor 26 to determine the particle mass and charge prior to step 206. In this example, as described above with respect to Figure 4A, the ion processing region 16 is illustratively implemented in the form of a conventional MS followed by a CDA or a CDMS, either of which are configured to measure the particle mass and charge directly in step 204 or to determine the particle mass and charge from measurements of charged particles made by the apparatus(es). However, it will be understood that the ion processing region 16 may alternatively be or include any apparatus or device, or combination of apparatus or devices, configured to measure the mass and charge of a particle or to measure one or more characteristics or properties of a particle that can determine or estimate both the mass and charge of the particle.In any case, once the particle mass and charge information has been determined in step 204, in step 206 the processor 26 operates to control the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 passes the charged particle exiting the ion processing region 16 towards the particle target 40 only if the particle's mass is within a specified particle mass range of 10-15 MDa and the particle's charge magnitude is within a specified particle charge magnitude range of 600-750e, otherwise controls the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 blocks the passage of the particle towards the target 40 or steers the charged particle away from the target 40.

[0052] Example 4

[0065] Referring now to Figure 4E, the urinary exome plot of Figure 4A is once again recreated with an example of the selection of yet another subpopulation or subset 400 of particles by the apparatus of Figure 1 for purification according to steps 204 and 206 of process 200 illustrated in Figure 3 superimposed thereon. As is evident from the plot of Figure 4A, the total population of the urinary exome appears to fall along a number of different oblique or graded subpopulations, subsets, or families, each of which is grouped with respect to or along a different constant mass-to-charge ratio (1 or greater) value or range. In this example, the selected subpopulation 400 is defined by a designated one of such mass-to-charge ratio (1 or greater) values ​​or ranges. In order for the processor 26 to control the voltage source VS3 in step 206 such that the charged particle deflector or steering device 18 passes only charged particles having a specified mass-to-charge ratio value or having a mass-to-charge ratio within a specified range of mass-to-charge ratio values ​​therethrough towards the particle target 40, the particle measurement information produced by the one or more apparatus or devices in the ion processing region 16 must include particle mass-to-charge ratio information or particle measurement information that allows the processor 26 to determine a mass-to-charge ratio prior to step 206. In this example, as described above with respect to Figure 4A, the ion processing region 16 is illustratively implemented in the form of a conventional MS followed by a CDA or a CDMS, either of which are configured to measure the mass and charge of the particles directly in step 204 or to determine the mass and charge of the particles from measurements of the charged particles made by the apparatus(es). However, it will be understood that the ion processing region 16 may alternatively be or include any apparatus or device, or combination of apparatus or devices, configured to measure the mass and charge of a particle or to measure one or more characteristics or properties of a particle that can determine or estimate both the mass and charge of the particle.In either case, the processor 26, in this embodiment, operates to calculate the mass-to-charge ratio of the particle as a function of the measured mass and charge of the particle.

[0053]

[0066] Once the particle mass-to-charge ratio information has been determined in step 204, in step 206 the processor 26 operates to control the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 passes the charged particle exiting the ion processing region 16 towards the particle target 40 only if the mass-to-charge ratio of the particle has a specified mass-to-charge ratio or has a mass-to-charge ratio within a specified mass-to-charge ratio value range, otherwise controls the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 blocks the passage of the particle towards the target 40 or steers the charged particle away from the target 40.

[0054]

[0067] In an alternative embodiment of the apparatus 10, the charged particle deflector or steering device 18 may be omitted and the ion processing region 16 may be implemented in the form of a conventional mass analyzer or mass-to-charge ratio filter, such as a quadrupole mass-to-charge ratio filter. In this embodiment, the charge of the particles need not be measured in the ion processing region 16 and therefore the one or more charge amplifiers CA may also be omitted. In this embodiment of the apparatus 10, step 206 of the process 200 may be omitted and the processor 26 in step 204 controls the voltage source VS2 such that the mass analyzer or mass-to-charge ratio filter passes therethrough only those charged particles having a mass-to-charge value within a selected mass-to-charge value range 400 towards the particle target 40.

[0055] Example 5

[0068] Referring now to Figure 4F, the urinary exome plot of Figure 4A is once again recreated with an example of a further selection of a further subpopulation or subset 402 of particles by the apparatus of Figure 1 for purification according to steps 204 and 206 of process 200 illustrated in Figure 3. In this example, as in Example 4, the selected subpopulation 402 is defined by a specified one of a number of different families of mass-to-charge ratios or mass-to-charge ratio ranges, and further defined by a specified range of mass values ​​between 10 MDa and 20 MDa. In order for the processor 26 to control the voltage source VS3 in step 206 such that the charged particle deflector or steering device 18 passes only charged particles having a specified mass-to-charge ratio or a mass-to-charge ratio within a specified mass-to-charge ratio range and having a mass value within a specified mass value range therethrough towards the particle target 40, the particle measurement information produced by the one or more apparatus or devices in the ion processing region 16 must include particle mass and mass-to-charge ratio information or particle measurement information that allows the processor 26 to determine the mass and mass-to-charge ratio of the particle prior to step 206. In this example, as described above with respect to Figure 4A, the ion processing region 16 is illustratively implemented in the form of a conventional MS followed by a CDA or a CDMS, either of which are configured to measure the mass and charge of the particle directly in step 204 or to determine the mass and charge of the particle from measurements of the charged particles made by the apparatus(es). However, it will be appreciated that ion processing region 16 may alternatively be or include any apparatus or device, or combination of apparatus or devices, configured to measure the mass and charge of a particle, or configured to measure one or more properties or characteristics of a particle that allow both the mass and charge of the particle to be determined or estimated. In either case, processor 26 operates to calculate the mass-to-charge ratio of the particle as a function of the measured mass and charge of the particle.

[0056]

[0069] Once the particle mass and mass-to-charge ratio information has been determined in step 204, in step 206 the processor 26 operates to control the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 passes the charged particle exiting the ion processing region 16 towards the particle target 40 only if the mass-to-charge ratio and mass of the particle are within a specified particle mass and mass-to-charge ratio range 402, otherwise the processor 26 controls the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 blocks the passage of the particle towards the target 40 or steers the charged particle away from the target 40.

[0057] Example 6

[0070] Referring now to Figure 4G, the urinary exome plot of Figure 4A is once again recreated with an example of a further selection of a further subpopulation or subset 404 of particles by the apparatus of Figure 1 for purification according to steps 204 and 206 of process 200 illustrated in Figure 3. In this example, similar to examples 4 and 5, the selected subpopulation 404 is defined by a specified one of a family of a plurality of different fixed mass-to-charge ratios or mass-to-charge ratio ranges, and further defined by a specified range of charge magnitude values ​​between 300e and 450e. In order for the processor 26 to control the voltage source VS3 in step 206 such that the charged particle deflector or steering device 18 passes therethrough towards the particle target 40 only those charged particles having a specified mass-to-charge ratio or having a mass-to-charge ratio within a specified mass-to-charge ratio range and having a charge magnitude value within a specified charge magnitude value range, the particle measurement information produced by the one or more apparatus or devices in the ion processing region 16 must include particle mass-to-charge ratio and charge magnitude information or particle measurement information that allows the processor 26 to determine the particle mass-to-charge ratio and charge magnitude prior to step 206. In this example, as described above with respect to Figure 4A, the ion processing region 16 is illustratively implemented in the form of a conventional MS followed by a CDA or a CDMS, either of which are configured to measure the particle mass and charge directly in step 204 or to determine the particle mass and charge from measurements of the charged particles made by the apparatus(es). However, it will be appreciated that ion processing region 16 may alternatively be or include any apparatus or device, or combination of apparatus or devices, configured to measure the mass and charge of a particle, or configured to measure one or more properties or characteristics of a particle that allow both the mass and charge of the particle to be determined or estimated. In either case, processor 26 operates to calculate the mass-to-charge ratio of the particle as a function of the measured mass and charge of the particle.

[0058]

[0071] Once the particle mass-to-charge ratio and charge magnitude information has been determined in step 204, in step 206 the processor 26 operates to control the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 passes the charged particle exiting the ion processing region 16 towards the particle target 40 only if the particle's mass-to-charge ratio and charge magnitude are within a specified particle mass-to-charge ratio and charge magnitude value range 404, otherwise the processor 26 controls the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 blocks the passage of the particle towards the target 40 or steers the charged particle away from the target 40.

[0059] Example 7

[0072] Referring now to Figure 4H, the urinary exome plot of Figure 4A is again reproduced with an example of a further selection of a further subpopulation or subset 406 of particles by the apparatus of Figure 1 for purification according to steps 204 and 206 of process 200 illustrated in Figure 3. In this example, similar to examples 4, 5 and 6, the selected subpopulation 406 is defined by a specified one of a family of a plurality of different fixed mass-to-charge ratios or mass-to-charge ratio ranges, and further defined by a specified range of mass values ​​between 15 MDa and 25 MDa, and a specified range of charge magnitude values ​​between 300e and 450e. In order for the processor 26 to control the voltage source VS3 in step 206 such that the charged particle deflector or steering device 18 passes therethrough towards the particle target 40 only those charged particles having a specified mass-to-charge ratio or having a mass-to-charge ratio within a specified mass-to-charge ratio range and having a charge magnitude value within a specified charge magnitude value range and a mass value within a specified mass value range, the particle measurement information produced by the one or more apparatus or devices in the ion processing region 16 must include mass-to-charge ratio and charge magnitude information or particle measurement information that allows the processor 26 to determine the mass, mass-to-charge ratio and charge magnitude of the particle prior to step 206. In this example, as described above with respect to Figure 4A, the ion processing region 16 is illustratively implemented in the form of a conventional MS followed by a CDA or a CDMS, either of which is configured to measure the mass and charge of the particle directly in step 204 or to determine the mass and charge of the particle from measurements of the charged particles made by the apparatus(es). However, it will be understood that the ion processing region 16 may alternatively be or include any apparatus or device, or combination of apparatus or devices, configured to measure the mass and charge of a particle or to measure one or more characteristics or properties of a particle that can determine or estimate both the mass and charge of the particle.In either case, the processor 26 is operative to calculate the mass-to-charge ratio of the particle as a function of the measured mass and charge of the particle.

[0060]

[0073] Once the particle mass-to-charge ratio and charge magnitude information has been determined in step 204, in step 206 the processor 26 operates to control the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 passes the charged particle exiting the ion processing region 16 towards the particle target 40 only if the particle's mass-to-charge ratio, charge magnitude, and mass are within a specified particle mass-to-charge ratio, charge magnitude, and mass value range 406, otherwise controls the voltage source VS3 so that the charged particle deflector or charged particle steering device 18 blocks the particle from passing towards the target 40 or steers the charged particle away from the target 40.

[0061]

[0074] Although sample 24 used in the example shown in FIGS. 4A - 4H is a urinary exosome, it will be understood that in other applications, sample 24 can be any material, whether essentially biological or not, whether liquid or otherwise. Additional examples of biological substances and materials that can be used as sample 24 can include, but are not limited to, other exosomes, endosomes, microvesicles in general, ectosomes, apoptotic bodies, retroviruses, exomeres, kyromicrons, DNA, RNA, proteins, lipids, acids, carbohydrates, enzymes, viruses, bacteria, etc. In some embodiments, purified (and in some cases, amplified) particles can be used for the research, assembly, and / or manufacture of gene therapy products and / or other products. Also, the example shown in FIGS. 4A - 4H illustrates a sub - population or subset of charged particles generated from sample 24 defined by various values or ranges of particle mass, charge, and / or mass - to - charge ratio, but it will be understood that the apparatus 10 illustrated in FIG. 1 and the processes 100, 200 for operating the apparatus 10 are not so limited. In particular, it will be understood that the apparatus 10 can be configured and the processes 100 and / or 200 can be modified to collect sub - populations or subsets of charged particles alternatively or additionally defined by values or ranges of particle mobility and / or other molecular properties. As one specific example without any limitation to consider, the ion processing region 16 can be configured to include an apparatus for measuring, and otherwise determining, the mass, charge, and mobility of particles, and various purified multi - dimensional sub - populations of charged particles can be defined by values and / or ranges of particle mass, mass - to - charge ratio, magnitude or state of charge, and mobility.

[0062] Example 8

[0075] Referring again to FIG. 5, process 500 was performed using a sample of bovine milk, where the specified type of particle in the bovine milk was an exosome. In step 502, pooled unprocessed (raw) bovine milk (from approximately 20 cows) was provided. Then, in step 504, the raw bovine milk sample was enriched for exosomes, as described below. Of the approximately 200 minutes of collected bovine milk sample, a 50 milliliter aliquot of the raw milk was skimmed and then further centrifuged to reduce the amount of apoptotic bodies in the sample. This was accomplished, by way of example, by centrifuging the raw milk at 2000×g for 10 minutes at 4° Celsius to remove the concentrated milk fat layer. The remaining suspension was sequestered and centrifuged at 12000×g for 20 minutes at 4° Celsius to remove cells and other debris. Acetic acid was then added to the supernatant (at a concentration of 1 volume percent) and mixed for 5 minutes to precipitate non-EV (non-extracellular vesicle) proteins, particularly casein, which has an isoelectric point of 4.6. The precipitate was separated by conventional centrifugation at 10,000×g for 10 minutes at 4°C. The resulting supernatant, a mixture of proteins, lipids, and other species, including EVs (called whey), was ultracentrifuged at 210,000×g for 70 minutes at 4°C. The resulting pellet was resolubilized in 500 microliters of 100 mM ammonium acetate and centrifuged at 10,000×g for 5 minutes at 4°C to remove remaining precipitate. The resulting exosome-containing EV supernatant was then diluted 100-fold with a solution of 100 mM ammonium acetate to form sample 24, from which ions were generated using the apparatus 10 of FIG. 1.

[0063]

[0076] The apparatus 10 was configured as described below by way of example, but it will be understood that the configuration of the apparatus 10 described below is merely one of several different possible configurations of the apparatus 10 illustrated in FIG. 1 and described above. For this particular example, the ion generator 22 is an electrospray ionization (EIS) device having a borosilicate capillary emitter of about 5 μm diameter, and an emitter potential of about 1.4 kV was used to generate ions from the concentrated sample 24. The electrosprayed ions are directed through a capillary interface into the source region of the apparatus 10, which is illustratively configured as a CDMS device described above. In this particular embodiment, the ion processing region 16 includes a hybrid ion funnel-ion carpet interface, such as that illustrated and described in co-pending International Application PCT / US2019 / 0132274, which is incorporated herein by reference, through which ions from the source capillary are directed. Following this interface, the ions were sent through an RF-only hexapole where the ions were made to collide, which thermalized the distribution of ion kinetic energies. As the ions exited the hexapole, they entered an RF-only quadrupole, which acts as a low-pass filter tuned to transmit larger ions with mass-to-charge (m / z) ratio values ​​above about 12000. Removal of the low m / z species ensured that the measurement time was optimized for the high m / z ions. A DC offset voltage of 100 V on the hexapole was used to set the nominal ion energy per charge. The ions were then collected into the entrance of a dual hemispherical deflection energy analyzer, and the energy-selected ions exiting the energy analyzer were then admitted to an electrostatic linear ion trap (ELIT) containing a charge detection cylinder.

[0064]

[0077] As each ion entered the ELIT, it induced a charge in the charge detection cylinder. At the start of collection and each trapping event, both end caps of the ELIT were in transmission mode, allowing the ion to travel through the trap. The trapping event was initiated by switching the rear end cap from transmission to trapping mode, reflecting the ion back through the charge detection cylinder to the entrance of the ELIT. Following a short delay of 0.3 ms, the front end cap was switched to trapping mode, and the trapped ion oscillated back and forth in the ELIT. After 100 ms, the trapping event was terminated and both end caps of the ELIT were switched back to transmission mode. After a 1 ms delay, the process was repeated for each of the ions. During the 100 ms measurement time, each ion oscillated through the charge detection cylinder of the ELIT, inducing a periodic signal that was amplified by a charge-sensitive preamplifier CA, digitized, and then analyzed using fast Fourier transform. The mass-to-charge ratio of the ions is derived from the fundamental frequency of the measurement, and the charge is derived from the magnitude of the fundamental frequency. The mass distribution was generated by multiplying the mass-to-charge ratio value by the measured charge for each ion and binning the resulting masses.

[0065]

[0078] In step 506, process 100 of FIG. 2 is performed as described above, and in step 106 of process 100, a scatter plot of the magnitude of charge (in units of elementary charge, e) of the charged particles versus the mass of the charged particles (in units of megadaltons, MDa), shown in FIG. 6A, is generated. The scatter plot is then processed in step 108 to determine a subpopulation of the plot data that can be identified as exosomes. In one embodiment of process 500, all particles having a mass greater than 9.8 megadaltons (commonly understood to be the minimum mass of an exosome) are considered to be exosomes, and are represented in FIG. 6A by the dashed vertical exosome mass threshold line EM THAll charged particles to the right of are so identified. In some embodiments, process 200 can be performed as described above, for example, to collect and / or purify the exosomes identified in step 108.

[0066]

[0079] In an alternative embodiment of process 500, the data of the scatter plot of FIG. 6A is processed in step 108 to determine one or more subpopulations of charged particles that can be identified as exosomes and / or to determine whether there are multiple subpopulations of charged particles that are distinguishable from each other, i.e., whether there are subpopulations that can be families of particles from the CDMS data. In this regard, in step 108, the processor 26 is programmed to perform a conventional two-dimensional Gaussian mixture model (GMM) to fit the two-dimensional mass-to-charge data of FIG. 6A, which assumes that the particle subpopulations are classified into families of related mass and charge, and that the distribution of these families follows a normal distribution. With this assumption, the processor 26 is programmed to perform a conventional clustering analysis on the charged particle data, the result of which is a distribution of multiple two-dimensional mass-to-charge subpopulations. When combined, the sum of these subpopulations captures the main features of the two-dimensional CDMS data. For simplicity, the number of possible subpopulations was limited to between 1 and 10 two-dimensional Gaussians. Except for this restriction, the analysis was unsupervised, and the algorithm determined the number of subpopulations that, when summed, best fit the two-dimensional CDMS data set, and the location and width associated with each subpopulation. For the CDMS data set shown in FIG. 6A, the analysis collapsed into a best-fit model constructed from six independent subpopulations S1-S6 shown in the example of FIG. 6B. Since each of the six subpopulations of charged particles illustrated in FIG. 6B are Gaussian distributed, it will be understood that the boundaries of each of S1-S6 are merely approximate and are included in FIG. 6B merely to indicate the location and approximate size of the subpopulations S1-S6 relative to one another. In another embodiment, one or more other conventional statistical models can be used to analyze the particle mass and charge data set produced by the CDMS 10.

[0067]

[0080] The data in FIG. 6B show that the lowest mass subpopulation S1 observed in sample 24 corresponds to a relatively narrow distribution centered at mass (m)=5.7±1.6 MDa and charge (z)=145±38e. This subpopulation S1 comprises approximately 27% (975 of 3586) of the total number of charged particles in this data set. The highest mass subpopulation S6 observed in sample 24 corresponds to a broad distribution centered at m=27.7±5.4 MDa and z=594±76e. This subpopulation S6 comprises approximately 22% (772 of 3586) of the total number of charged particles in this data set. The S2 subpopulation (or family) centered at m=10.2±1.9 MDa and z=189±44e comprises only 3% of the total number of charged particles in this data set, making it the least abundant subpopulation. The S3 (m=12.5±2.9MDa, z=296±31e) subpopulation, which contains about 4% of the total number of charged particles, and the S4 (m=17.6±2.6MDa, z=488±76e) subpopulation, which contains about 18% of the total number of charged particles, are substantially better determined on the basis of charge than mass. This implies that these subpopulations or families consist of similarly sized particles that differ substantially at the molecular level, thus influencing the charge of each particle more than the mass of each particle. The S5 (m=23.4±3.4MDa, z=550±113e) subpopulation, which contains about 26% of the total number of charged particles, appears to be better determined by mass than by charge dimensions, indicating that they are more similar in charge properties than size.

[0068]

[0081] From the Gaussian model cluster analysis undertaken in step 108 of the process 100 described herein, it can be estimated which fraction of particles in the dataset are exosomes. With respect to the average mass of the various subpopulations, only the S1 subpopulation (m=5.7±1.6 MDa) is too small to be exosomes (based on the smallest exosome mass being about 9.8 MDa). Since S1 represents 27% of the total number of charged particles in the dataset, the remaining 73% of the charged particles in the dataset are within the mass range considered to be exosomes. In some embodiments, the process 200 can be performed as described above to collect and / or purify the exosomes identified in this embodiment of step 108, as an example.

[0069]

[0082] While the invention has been illustrated and described in detail in the drawings and description above, it is understood that this is to be considered as illustrative and not limiting in nature, and that only exemplary embodiments thereof have been shown and described, and that protection is desired for all changes and modifications that fall within the spirit of the invention. For example, in some embodiments where the ion processing region 16 is implemented in the form of a CDMS, the charge detector of the CDMS is controlled, for example, by selective control of voltage source VS2, to selectively release charged particles or to prevent release of particles therefrom. In embodiments of the CDMS where the charge detector is, for example, an electrostatic linear ion trap (ELIT), the voltage applied by voltage source VS2 to one or both of its end caps is controlled, for example, to allow ions trapped and oscillating therein to exit the ELIT in the direction of particle collection target 40, or to destabilize the oscillation of ions within the ELIT so that they contact structures therein and thus prevent ions from being released from the ELIT. In such embodiments, such control of the ELIT may obviate the need for a charged particle deflector or charged particle steering device 18, such that it may be omitted. In an embodiment of the CDMS in which the charge detector is an Orbitrap, voltage source VS2 can be similarly controlled to have the same effect.

Claims

1. 1. A particle purification device comprising: an ion generator configured to generate charged particles from the sample; an ion processing region configured to receive the charged particles generated by the ion generator and measure at least one of a mass and a charge magnitude of the generated charged particles; a particle collection target having a collection surface; an ion trap disposed between the particle collection target and a charged particle deflection or steering device; a means for selectively passing charged particles exiting the ion processing region into the ion trap, comprising: a processor-controllable charged particle deflection or steering device for selectively passing charged particles into the ion trap; the processor; a memory storing instructions executable by the processor, (i) controlling, by the processor, the charged particle deflection or steering device to pass each of the measured charged particles having at least one of: (a) a measured mass equal to or within a selected particle mass range; (b) a measured charge magnitude equal to or within a selected charge magnitude range; and (c) a mass to charge ratio equal to or within a selected mass to charge ratio range into the ion trap for trapping in the ion trap; and (ii) controlling the ion trap by the processor to release charged particles trapped in the ion trap and accelerate the charged particles towards the collection surface of the particle collection target for collection; a memory storing instructions for causing the 16. A particle purification device comprising:

2. 2. The particle purification device of claim 1, wherein the instructions stored in the memory further include instructions executable by the processor to cause the processor to control the charged particle deflection or steering device to otherwise block the passage of the measured charged particles to the particle collection target.

3. 3. The particle purification device of claim 1 or claim 2, wherein the ion processing region includes a charge detection mass analyzer configured to receive the charged particles generated by the ion generator and to measure the mass and charge magnitude of the generated charged particles.

4. 3. The particle purification device of claim 1 or claim 2, comprising at least one of a mass analyzer, a mass spectrometer, and a mass to charge ratio filter configured to pass charged particles having a mass to charge ratio that is a selected mass to charge ratio or that is within a selected range of mass to charge ratios, followed by a charge detection array configured to measure a magnitude of charge of charged particles exiting the one of the mass analyzer, the mass spectrometer, and the mass to charge ratio filter.

5. 1. A method for purifying particles, comprising the steps of: Producing charged particles from a sample; Measuring at least one of the mass and charge of the generated charged particles; selectively passing, by a charged particle deflection or steering device, each of the measured charged particles having at least one of: (a) a measured mass equal to or within a selected particle mass range; (b) a measured charge magnitude equal to or within a selected charge magnitude range; and (c) a mass-to-charge ratio equal to or within a selected mass-to-charge ratio range, into the ion trap for trapping therein; controlling the ion trap to release charged particles trapped in the ion trap and accelerate the charged particles towards a collection surface of a particle collection target for collection; A method comprising:

6. 6. The method of claim 5, further comprising collecting at the collection surface of the particle collection target the measured charged particles selectively passed thereto.

7. The method of claim 6 further comprising harvesting the charged particles collected at the collection surface of the particle collection target.

8. 8. The method of claim 7, further comprising amplifying the collected charged particles.

9. 10. The particle purification device of claim 1, wherein the sample contains exosomes, The instructions stored in the memory further include instructions executable by the processor to determine, from the measured masses of at least some of the generated charged particles, a subset of the generated charged particles having masses that identify corresponding generated charged particles that are exosome ions; The instructions stored in the memory further comprise instructions executable by the processor to control the charged particle deflection or steering device to pass the exosome ions through the ion trap. Particle purification device.

10. 6. The method of claim 5, wherein generating charged particles from the sample comprises generating charged particles from a sample that contains exosomes; said measuring includes at least measuring a mass of said charged particle; The method further includes identifying a subset of the generated charged particles that are exosome ions from the measured masses of at least some of the generated charged particles; The method, wherein said selectively passing comprises passing said exosome ions through said ion trap.

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