DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Specification
The disclosure is objected to because of informalities indicated in an attached, marked-up copy of the specification showing tracking of changes.
Appropriate correction is required.
Claim Objections
Claims 1, 24 and 28 objected to because of the following informalities:
In Claim 1, line 11 will be read without the comma (“,”) at the end of the line.
In Claim 24, line 2 will be read without the comma (“,”) at the end of the line.
In Claim 28, lines 11-14 will be read as “a detector configured to detect one or more detectable signals produced in the detection zone when the fluid handling apparatus effects the flow of the population of labeled nanoparticles [[
Appropriate correction is required.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
Determining the scope and contents of the prior art.
Ascertaining the differences between the prior art and the claims at issue.
Resolving the level of ordinary skill in the pertinent art.
Considering objective evidence present in the application indicating obviousness or non-obviousness.
Claim(s) 1-4, 9, 12, and 19-20 is/are rejected under 35 U.S.C. 103 as being unpatentable over Wang et al. (US 2013/0167623 A1) in view of Nechev et al. (US 2019/0269795 A1).
Regarding independent Claim 1, Wang discloses a method of characterizing a nanoparticle in a population of labeled nanoparticles, the method comprising:
determining a retention time taken for the nanoparticle to flow from a first point in or proximal to a fluidic channel to or through a detection zone (Figure 1; [0029] “determine a time that each of the plurality of particles passes through the detection zone 18”) of the fluidic channel (Figure 1; [0029] “a detection zone 18 of the fluid channel 14”) when a fluidic sample that comprises the population of labeled nanoparticles ([0029] “a plurality of particles in a fluid”) flows through the fluidic channel (Figure 1; [0029] “a fluid channel 14”) to produce retention data (Table 2: Column 2 shows Retention Time);
determining a size measure of the nanoparticle from the retention data to produce nanoparticle size data (Figure 1; [0029] “a data processing system 22 … configured to determine a size of each of the plurality of particles based on the time that each of the plurality of particles passes through the detection zone 18”); and
detecting a detectable signal produced (Figure 1; [0029] “a detection system 20 arranged to detect each of the plurality of particles based on corresponding responses to the illumination light”, wherein “responses to the illumination light” are interpreted as signals produced) by one or more labels of one or more components of the nanoparticle (Figure 5C; [0056] “Each spike in the raw APD fluorescence data (FIG. 5C) represents a single DNA molecule”) when the nanoparticle flows through the detection zone (Figure 1; [0029] “each of the plurality of particles passes through the detection zone 18”) of the fluidic channel (Figure 1; [0029] “a detection zone 18 of the fluid channel 14”) to produce signal data (Figures 5A-5C), but does not specifically teach:
determining at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data, thereby characterizing the nanoparticle in the population of labeled nanoparticles.
However, Nechev, in the same field of lipid nanoparticles effective to deliver a nucleic acid payload, teaches determining at least one payload property of the nanoparticle ([0007] “a formulation comprising lipid nanoparticles comprising an RNAi agent payload”) from the signal data to produce nanoparticle payload property data ([0070] “The total RNA in the formulation can be determined by the signal from the sample containing the surfactant, relative to a standard curve”), thereby characterizing the nanoparticle in the population of labeled nanoparticles (it is interpreted that once a nanoparticle is determined to comprise or not comprise an agent payload, said nanoparticle can be characterized).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the method of Wang with the teachings of Nechev, for determining at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data, thereby characterizing the nanoparticle in the population of labeled nanoparticles, because “there remains a need for improved processes and apparatuses for the manufacture of lipid nanoparticles capable of carrying a therapeutic payload. The present invention provides such methods, processes and systems for the manufacture of lipid nanoparticles which sufficiently encapsulate a nucleic acid payload, specifically RNAi agents, for delivery to mammalian cells.” (Nechev, [0005])
Regarding Claim 2, modified Wang discloses the method of claim 1, wherein the first point in or proximal to the fluidic channel comprises an inlet (Figure 1; [0030] “an upper portion 30 and a lower portion 32 that fasten together to contain a sample and sample holder 34 therein” such that fluid channel 14 has an inlet reaching into sample holder 34) to the fluidic channel (Figure 1; [0029] “a fluid channel 14”).
Regarding Claim 3, modified Wang discloses the method of claim 1, wherein the population of labeled particles comprises lipid particles ([0077] “separation of other biomolecules, such as proteins, peptides, RNA, lipids, vesicles, and organelles, can also be performed”; [Claim 3] “plurality of particles comprises at least one of polymer, pharmaceutical, fluorophore, DNA, RNA, lipid, emulsion, carbohydrate, metabolite, antibody, or protein molecules, vesicle or cells”), but does not specifically teach that the population of labeled nanoparticles comprises lipid nanoparticles.
However, Nechev, in the same field of lipid nanoparticles effective to deliver a nucleic acid payload, teaches that the population of labeled nanoparticles comprises lipid nanoparticles ([0007] “a formulation comprising lipid nanoparticles”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the method of Wang with the teachings of Nechev, wherein the population of labeled nanoparticles comprises lipid nanoparticles, because using labeled lipid nanoparticles to deliver payloads like mRNA or drugs allows researchers to track, visualize, and quantify how the delivery system moves through cells and living organisms.
Regarding Claim 4, modified Wang discloses the method of claim 1, but does not specifically teach that at least two, at least three, or more components of the nanoparticles in the population of nanoparticles comprise different labels from one another.
However, Nechev, in the same field of lipid nanoparticles effective to deliver a nucleic acid payload, teaches that at least two ([0074] “5′-end Cy-3 and Cy-5.5 (fluorophore) labeled RNAi agents”, wherein a “5′-end Cy-3” modification attaches a bright, yellow-orange fluorescent cyanine dye (excitation ~550 nm, emission ~564 nm) to the 5′ terminus of an oligonucleotide like DNA or RNA; “Cy-5.5” is a synthetic far-red and near-infrared (NIR) fluorescent dye used to label proteins, peptides, and nucleic acids), at least three, or more components of the nanoparticles in the population of nanoparticles comprise different labels from one another.
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the method of Wang with the teachings of Nechev, wherein at least two, at least three, or more components of the nanoparticles in the population of nanoparticles comprise different labels from one another, because using two or more different fluorescent labels in nanoparticles helps track delivery and drug release at the same time.
Regarding Claim 9, modified Wang discloses the method of claim 1, but does not specifically teach that at least a first component of the nanoparticles in the population of labeled nanoparticles comprises a payload molecule.
However, Nechev, in the same field of lipid nanoparticles effective to deliver a nucleic acid payload, teaches that at least a first component of the nanoparticles in the population of labeled nanoparticles comprises a payload molecule ([0007] “a formulation comprising lipid nanoparticles comprising an RNAi agent payload”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the method of Wang with the teachings of Nechev, wherein at least a first component of the nanoparticles in the population of labeled nanoparticles comprises a payload molecule, because using nanoparticles to carry a payload molecule like a drug, protein, or DNA/RNA protects the fragile cargo from breaking down in the body, helps it dissolve in blood, and delivers it directly to specific cells (like cancer cells) while keeping healthy tissues safe.
Regarding Claim 12, modified Wang discloses the method of claim 1, wherein at least a second component of the nanoparticles in the population of labeled nanoparticles comprises a molecule selected from the group consisting of: a polymer, a copolymer, a lipid, a fluorophore, a carbohydrate, an emulsion, a vesicle, a cell, polyethylene glycol (PEG), cholesterol, a liposome, a carbon nanotube, silica, and gold ([0077] “separation of other biomolecules, such as proteins, peptides, RNA, lipids, vesicles, and organelles, can also be performed”; [Claim 3] “plurality of particles comprises at least one of polymer, pharmaceutical, fluorophore, DNA, RNA, lipid, emulsion, carbohydrate, metabolite, antibody, or protein molecules, vesicle or cells”).
Regarding Claim 19, modified Wang discloses the method of claim 1, comprising distinguishing nanoparticles comprising a payload molecule from nanoparticles lacking a payload molecule in the population of the labeled nanoparticles ([0078] “capillaries or microfluidic channels may be surface functionalized with moieties that interact with the separated molecules to enhance the distinction between molecules beyond size alone”) using at least the signal data (Figures 5A-5C).
Regarding Claim 20, modified Wang discloses the method of claim 1, comprising determining a distribution of the at least one payload property of the nanoparticles in the population of labeled nanoparticles (Figures 8A-8B; [0060] “time domain data in FIG. 8A is remapped into a spatial domain heat map in FIG. 8B to illustrate the distribution of molecules”).
Claim 21 is rejected under 35 U.S.C. 103 as being unpatentable over Wang et al. (US 2013/0167623 A1) and Nechev et al. (US 2019/0269795 A1) as applied to claim 1 above, and further in view of Chiu et al. (US 2021/0016276 A1).
Regarding Claim 21, modified Wang discloses the method of claim 1, but does not specifically teach determining a concentration of nanoparticles in the population of labeled nanoparticles using at least the signal data.
However, Chiu, in the same field of performing analyses of biological nanoparticles, teaches determining a concentration of nanoparticles in the population of labeled nanoparticles using at least the signal data ([0500] “FIG. 23C and FIG. 23D show examples of the detected signal intensity distributions for the ANEPPS labeled exosomes (FIG. 23C) and gold nanoparticles (FIG. 23D), respectively. FIG. 23C depicts an example signal intensity histogram from the membrane dye-labeled vesicles (n=636 events). FIG. 23D shows an example signal intensity histogram from the gold nanoparticles (n=538 events) measured together with membrane-dye labeled exosomes in the same experiment at the same time … The gold nanoparticles also can serve as a concentration standard: by comparing the spike (detected photon burst) frequency between the vesicle sample and the known concentration of the gold nanoparticles, the concentration of the vesicles in the sample can be determined”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the method of Wang with the teachings of Chiu, for determining a concentration of nanoparticles in the population of labeled nanoparticles using at least the signal data, because determining the concentration of labeled nanoparticles helps confirm delivery accuracy, track target tracking in cells, and ensure safe, repeatable results in medicine and research.
Claim 24 is rejected under 35 U.S.C. 103 as being unpatentable over Steinmiller et al. (US 2016/0305878 A1) in view of Wang et al. (US 2013/0167623 A1) and Nechev et al. (US 2019/0269795 A1).
Regarding independent Claim 24, Steinmiller discloses a kit, comprising:
a device (Figure 1C; [0045] “fluidic device 10”) comprising a fluidic channel (Figure 1C; [0035] “a plurality of channel segments (first 26, second 28, and third 30)”) having a detection zone (Figure 1C; [0045] “fluidic device 10 is positioned between a light source 36 and an optical detector 38 such that first side 20 (comprising one or more channel segments) faces the detector and second side 22 (comprising one or more optical elements) faces light source 36 and is exposed to light 42. The detector may be associated with one or more fluidic channel segments in the fluidic device, e.g., to determine light transmission through one or more of the channel segments”, whereby “one or more fluidic channel segments” are interpreted as detection zones); and
instructions for using the device ([0099] “a set of instructions for use of the kit”), but does not specifically teach to:
determine a retention time taken for a nanoparticle to flow from a first point in or proximal to the fluidic channel to or through the detection zone of the fluidic channel when a fluidic sample that comprises a population of labeled nanoparticles flows through the fluidic channel to produce retention data;
determine a size measure of the nanoparticle from the retention data to produce nanoparticle size data;
detect a detectable signal produced by one or more labels of one or more components of the nanoparticle when the nanoparticle flows through the detection zone of the fluidic channel to produce signal data; and
determine at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data.
However, Wang, in the same field of flow cytometry, teaches to:
determine a retention time taken for a nanoparticle to flow from a first point in or proximal to the fluidic channel to or through the detection zone (Figure 1; [0029] “determine a time that each of the plurality of particles passes through the detection zone 18”) of the fluidic channel (Figure 1; [0029] “a detection zone 18 of the fluid channel 14”) when a fluidic sample that comprises a population of labeled nanoparticles ([0029] “a plurality of particles in a fluid”) flows through the fluidic channel (Figure 1; [0029] “a fluid channel 14”) to produce retention data (Table 2: Column 2 shows Retention Time);
determine a size measure of the nanoparticle from the retention data to produce nanoparticle size data (Figure 1; [0029] “a data processing system 22 … configured to determine a size of each of the plurality of particles based on the time that each of the plurality of particles passes through the detection zone 18”); and
detect a detectable signal produced (Figure 1; [0029] “a detection system 20 arranged to detect each of the plurality of particles based on corresponding responses to the illumination light”, wherein “responses to the illumination light” are interpreted as signals produced) by one or more labels of one or more components of the nanoparticle (Figure 5C; [0056] “Each spike in the raw APD fluorescence data (FIG. 5C) represents a single DNA molecule”) when the nanoparticle flows through the detection zone (Figure 1; [0029] “each of the plurality of particles passes through the detection zone 18”) of the fluidic channel (Figure 1; [0029] “a detection zone 18 of the fluid channel 14”) to produce signal data (Figures 5A-5C).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the kit of Steinmiller with the teachings of Wang, to determine a retention time taken for a nanoparticle to flow from a first point in or proximal to the fluidic channel to or through the detection zone of the fluidic channel when a fluidic sample that comprises a population of labeled nanoparticles flows through the fluidic channel to produce retention data; determine a size measure of the nanoparticle from the retention data to produce nanoparticle size data; and detect a detectable signal produced by one or more labels of one or more components of the nanoparticle when the nanoparticle flows through the detection zone of the fluidic channel to produce signal data, because “CICS has a sheet-like observation volume that enables substantially 100% detection efficiency of single molecules within the separation capillary in contrast to standard laser-induced fluorescence (LIF).” (Wang, [0026])
Steinmiller is also silent with respect to: determine at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data.
However, Nechev, in the same field of lipid nanoparticles effective to deliver a nucleic acid payload, teaches to determine at least one payload property of the nanoparticle ([0007] “a formulation comprising lipid nanoparticles comprising an RNAi agent payload”) from the signal data to produce nanoparticle payload property data ([0070] “The total RNA in the formulation can be determined by the signal from the sample containing the surfactant, relative to a standard curve”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the kit of Steinmiller with the teachings of Nechev, to determine at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data, because “there remains a need for improved processes and apparatuses for the manufacture of lipid nanoparticles capable of carrying a therapeutic payload. The present invention provides such methods, processes and systems for the manufacture of lipid nanoparticles which sufficiently encapsulate a nucleic acid payload, specifically RNAi agents, for delivery to mammalian cells.” (Nechev, [0005])
Claim 25 is rejected under 35 U.S.C. 103 as being unpatentable over Steinmiller et al. (US 2016/0305878 A1) and Wang et al. (US 2013/0167623 A1) and Nechev et al. (US 2019/0269795 A1) as applied to claim 24 above, and further in view of Chiu et al. (US 2021/0016276 A1).
Regarding Claim 25, modified Steinmiller discloses the kit of claim 24, further comprising instructions for using the device (see claim 24 rejection), but does not specifically teach instructions for using the device to determine a concentration of nanoparticles in the population of labeled nanoparticles using at least the signal data.
However, Chiu, in the same field of performing analyses of biological nanoparticles, teaches instructions for using the device ([0065] “the kit further comprises instructions”) to determine a concentration of nanoparticles in the population of labeled nanoparticles using at least the signal data ([0500] “FIG. 23C and FIG. 23D show examples of the detected signal intensity distributions for the ANEPPS labeled exosomes (FIG. 23C) and gold nanoparticles (FIG. 23D), respectively. FIG. 23C depicts an example signal intensity histogram from the membrane dye-labeled vesicles (n=636 events). FIG. 23D shows an example signal intensity histogram from the gold nanoparticles (n=538 events) measured together with membrane-dye labeled exosomes in the same experiment at the same time … The gold nanoparticles also can serve as a concentration standard: by comparing the spike (detected photon burst) frequency between the vesicle sample and the known concentration of the gold nanoparticles, the concentration of the vesicles in the sample can be determined”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the kit of Steinmiller with the teachings of Chiu, further comprising instructions for using the device to determine a concentration of nanoparticles in the population of labeled nanoparticles using at least the signal data, because determining the concentration of labeled nanoparticles helps confirm delivery accuracy, track target tracking in cells, and ensure safe, repeatable results in medicine and research.
Claim(s) 28, 33-34, 37, 39, 44-45 and 49 is/are rejected under 35 U.S.C. 103 as being unpatentable over Chiu et al. (US 2021/0016276 A1) in view of Wang et al. (US 2013/0167623 A1) and Nechev et al. (US 2019/0269795 A1).
Regarding independent Claim 28, Chiu discloses a system for characterizing a nanoparticle in a population of labeled nanoparticles, comprising:
a device receiving area (Figure 17A: region between Sample channel (1) and Outlet channel (2) is interpreted as receiving area) configured to receive a device comprising a fluidic channel (Figure 17A; [0471] “a microfluidic chip”) having a detection zone ([0009] “the detection region is within the at least one constriction”; Figure 17A; [0471] “The microfluidic channel was fabricated to have a 2 μm-wide and 75 μm-long constriction”; [0471] “The length of the illumination laser line was around 20 μm for the 633 nm laser (FIG. 17A). The 633 nm laser line was made to be ten times longer than the width of the channel”, wherein the overlap of “the illumination laser line” and “the width of the channel” is the detection zone);
a fluid handling apparatus configured to effect a flow of a fluidic sample (Figure 17A; [0471] “For sample injection and flow, two holes were punched having approximately 1-2 mm diameter; hole 1 was used for loading approximately 5 μL of sample, and hole 2 was left empty as the waste outlet. Prior to loading the sample, the channel was first filled with buffer, after which residual solutions in both reservoirs were withdrawn; upon pipetting approximately 5 μL of sample solution, flow was readily initiated without external pump because of the height difference (approximately 5-10 mm) in the fluid level between the two reservoirs, which made operation simple”) that comprises the population of labeled nanoparticles ([0473] “the copy number of biomarkers on individual nanoparticles”) through the detection zone ([0009] “the detection region is within the at least one constriction”; Figure 17A; [0471] “The microfluidic channel was fabricated to have a 2 μm-wide and 75 μm-long constriction”; [0471] “The length of the illumination laser line was around 20 μm for the 633 nm laser (FIG. 17A). The 633 nm laser line was made to be ten times longer than the width of the channel”, wherein the overlap of “the illumination laser line” and “the width of the channel” is the detection zone) when the fluid handling apparatus (Figure 17A; [0471] “For sample injection and flow, two holes were punched having approximately 1-2 mm diameter; hole 1 was used for loading approximately 5 μL of sample, and hole 2 was left empty as the waste outlet”) is operably connected to the device (Figure 17A; [0471] “a microfluidic chip”) and when the fluidic sample is disposed in the fluidic channel (implicit for sample to be present in microfluidic channel when sample flow is initiated; [0471]);
a light source configured to introduce an incident light toward the detection zone (Figure 17A; [0471] “The 633 nm laser line was made to be ten times longer than the width of the channel so that the laser illumination was homogeneous across the width of the channel”) when the device (Figure 17A; [0471] “a microfluidic chip”) is received in the device receiving area (Figure 17A: region between Sample channel (1) and Outlet channel (2) is interpreted as receiving area);
a detector configured to detect one or more detectable signals produced (Figure 17A; [0472] “In addition to the two APD detectors used to detect fluorescence excited by the 633 nm and 405 nm lasers, a third APD is used to detect the back scattered light from the Au nanoparticles for use as an internal standard to calibrate the detected light intensities from the bionanoparticles”) in the detection zone ([0009] “the detection region is within the at least one constriction”; Figure 17A; [0471] “The microfluidic channel was fabricated to have a 2 μm-wide and 75 μm-long constriction”; [0471] “The length of the illumination laser line was around 20 μm for the 633 nm laser (FIG. 17A). The 633 nm laser line was made to be ten times longer than the width of the channel”, wherein the overlap of “the illumination laser line” and “the width of the channel” is the detection zone) when the fluid handling apparatus effects the flow of the population of labeled nanoparticles through the fluidic channel (Figure 17A; [0471] “For sample injection and flow, two holes were punched having approximately 1-2 mm diameter; hole 1 was used for loading approximately 5 μL of sample, and hole 2 was left empty as the waste outlet. Prior to loading the sample, the channel was first filled with buffer, after which residual solutions in both reservoirs were withdrawn; upon pipetting approximately 5 μL of sample solution, flow was readily initiated without external pump because of the height difference (approximately 5-10 mm) in the fluid level between the two reservoirs, which made operation simple”; [0473] “the copy number of biomarkers on individual nanoparticles”) and when the device (Figure 17A; [0471] “a microfluidic chip”) is received in the device receiving area (Figure 17A: region between Sample channel (1) and Outlet channel (2) is interpreted as receiving area);
a controller ([0299] “a computer, controller, chip with integrated circuits, circuit board, electronic element, software, and/or algorithm”) that comprises, or is capable of accessing, computer readable media ([0411] “the computer includes a storage subsystem that provides a computer-readable storage medium”) comprising non-transitory computer-executable instructions ([0411] “the storage subsystem stores software (programs, code modules, instructions)”; [0415] “A file storage subsystem provides a non-transitory persistent (non-volatile) storage”) which, when executed by at least one electronic processor ([0411] “software modules or instructions can be executed by one or more processors”), perform at least:
flowing the fluidic sample through the fluidic channel using the fluid handling apparatus (Figure 17A; [0471] “For sample injection and flow, two holes were punched having approximately 1-2 mm diameter; hole 1 was used for loading approximately 5 μL of sample, and hole 2 was left empty as the waste outlet. Prior to loading the sample, the channel was first filled with buffer, after which residual solutions in both reservoirs were withdrawn; upon pipetting approximately 5 μL of sample solution, flow was readily initiated without external pump because of the height difference (approximately 5-10 mm) in the fluid level between the two reservoirs, which made operation simple”) such that the population of labeled nanoparticles ([0473] “the copy number of biomarkers on individual nanoparticles”) flow through the detection zone of the fluidic channel ([0009] “the detection region is within the at least one constriction”; Figure 17A; [0471] “The microfluidic channel was fabricated to have a 2 μm-wide and 75 μm-long constriction”; [0471] “The length of the illumination laser line was around 20 μm for the 633 nm laser (FIG. 17A). The 633 nm laser line was made to be ten times longer than the width of the channel”, wherein the overlap of “the illumination laser line” and “the width of the channel” is the detection zone);
introducing the incident light from the light source toward the detection zone (Figure 17A; [0471] “The 633 nm laser line was made to be ten times longer than the width of the channel so that the laser illumination was homogeneous across the width of the channel”) when the device (Figure 17A; [0471] “a microfluidic chip”) is received in the device receiving area (Figure 17A: region between Sample channel (1) and Outlet channel (2) is interpreted as receiving area); and
detecting a detectable signal produced by one or more labels of one or more components of the nanoparticle (Figure 17A; [0472] “In addition to the two APD detectors used to detect fluorescence excited by the 633 nm and 405 nm lasers, a third APD is used to detect the back scattered light from the Au nanoparticles for use as an internal standard to calibrate the detected light intensities from the bionanoparticles”) when the nanoparticle flows through the detection zone of the fluidic channel ([0009] “the detection region is within the at least one constriction”; Figure 17A; [0471] “The microfluidic channel was fabricated to have a 2 μm-wide and 75 μm-long constriction”; [0471] “The length of the illumination laser line was around 20 μm for the 633 nm laser (FIG. 17A). The 633 nm laser line was made to be ten times longer than the width of the channel”, wherein the overlap of “the illumination laser line” and “the width of the channel” is the detection zone) to produce signal data (Figure 17B), but does not specifically teach:
determining a retention time taken for the nanoparticle to flow from a first point in or proximal to a fluidic channel to or through the detection zone of the fluidic channel when the population of labeled nanoparticles flows through the fluidic channel to produce retention data;
determining a size measure of the nanoparticle from the retention data to produce nanoparticle size data; and
determining at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data to thereby characterize the nanoparticle in the population of labeled nanoparticles.
However, Wang, in the same field of flow cytometry, teaches:
determining a retention time taken for the nanoparticle to flow from a first point in or proximal to a fluidic channel to or through the detection zone (Figure 1; [0029] “determine a time that each of the plurality of particles passes through the detection zone 18”) of the fluidic channel (Figure 1; [0029] “a detection zone 18 of the fluid channel 14”) when the population of labeled nanoparticles ([0029] “a plurality of particles in a fluid”) flows through the fluidic channel (Figure 1; [0029] “a fluid channel 14”) to produce retention data (Table 2: Column 2 shows Retention Time); and
determining a size measure of the nanoparticle from the retention data to produce nanoparticle size data (Figure 1; [0029] “a data processing system 22 … configured to determine a size of each of the plurality of particles based on the time that each of the plurality of particles passes through the detection zone 18”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the system of Chiu with the teachings of Wang, for determining a retention time taken for the nanoparticle to flow from a first point in or proximal to a fluidic channel to or through the detection zone of the fluidic channel when the population of labeled nanoparticles flows through the fluidic channel to produce retention data; and determining a size measure of the nanoparticle from the retention data to produce nanoparticle size data, because “CICS has a sheet-like observation volume that enables substantially 100% detection efficiency of single molecules within the separation capillary in contrast to standard laser-induced fluorescence (LIF).” (Wang, [0026])
Chiu is also silent with respect to: determining at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data to thereby characterize the nanoparticle in the population of labeled nanoparticles.
However, Nechev, in the same field of lipid nanoparticles effective to deliver a nucleic acid payload, teaches determining at least one payload property of the nanoparticle ([0007] “a formulation comprising lipid nanoparticles comprising an RNAi agent payload”) from the signal data to produce nanoparticle payload property data ([0070] “The total RNA in the formulation can be determined by the signal from the sample containing the surfactant, relative to a standard curve”) to thereby characterize the nanoparticle in the population of labeled nanoparticles (it is interpreted that once a nanoparticle is determined to comprise or not comprise an agent payload, said nanoparticle can be characterized).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the system of Chiu with the teachings of Nechev, for determining at least one payload property of the nanoparticle from the signal data to produce nanoparticle payload property data to thereby characterize the nanoparticle in the population of labeled nanoparticles, because “there remains a need for improved processes and apparatuses for the manufacture of lipid nanoparticles capable of carrying a therapeutic payload. The present invention provides such methods, processes and systems for the manufacture of lipid nanoparticles which sufficiently encapsulate a nucleic acid payload, specifically RNAi agents, for delivery to mammalian cells.” (Nechev, [0005])
Regarding Claim 33, modified Chiu discloses the system of claim 28, wherein the light source comprises a cylindrical illumination apparatus (Figure 17A; [0471] “a line-confocal detection system for the high-sensitivity and high-throughput detection of fluorescence”).
Regarding Claim 34, modified Chiu discloses the system of claim 28, wherein at least two, at least three, or more components of the nanoparticles in the population of nanoparticles comprise different labels from one another ([0183] “the biological nanoparticle is associated with a detectable agent. In certain embodiments, the detectable agent is a luminescent dye, a fluorescent dye, a fluorescently labeled antibody, a fluorescently labeled protein, a fluorescently labeled nucleic acid, a fluorescently labeled lipid, a fluorescently labeled carbohydrate, a fluorescently labeled small molecule, a membrane dye, a fluorogenic dye, a dye, a polymer dot, a fluorogenic substrate of an enzyme, or a combination thereof”).
Regarding Claim 37, modified Chiu discloses the system of claim 28, wherein the non-transitory computer-executable instructions which, when executed by the electronic processor, further perform at least: determining a concentration of nanoparticles in the population of labeled nanoparticles using at least the signal data ([0020] “the concentration of the sample is determined by counting a spike frequency of the sample”).
Regarding Claim 39, modified Chiu discloses the system of claim 28, wherein the non-transitory computer-executable instructions which, when executed by the electronic processor, further perform at least: producing the nanoparticle size data ([0035] “a computer with software for: ranking biological nanoparticles based on the presence or absence of an emitted detectable light intensity of the biological nanoparticles; and measuring a size value of biological nanoparticles based on the emitted detectable light intensity of the biological nanoparticles”) and the nanoparticle payload property data substantially simultaneously (it has been held that a “recitation with respect to the manner in which a claimed apparatus is intended to be employed does not differentiate the claimed apparatus from a prior art apparatus” if the prior art apparatus teaches all the structural limitations of the claim. Ex parte Masham, 2 USPQ2d 1647 (1987)).
Regarding Claim 44, modified Chiu discloses the system of claim 28, wherein the non-transitory computer-executable instructions which, when executed by the electronic processor, further perform at least: illuminating the detection zone of the fluidic channel substantially uniformly across an entire cross section of the fluidic channel (Figure 17A; [0471] “The 633 nm laser line was made to be ten times longer than the width of the channel so that the laser illumination was homogeneous across the width of the channel”) such that each of the nanoparticles in the population of labeled nanoparticles passes through illumination light upon passing through the detection zone ([0471] “this was important for single-molecule counting to minimize any variability in detection sensitivity as the molecules pass through the channel at different lateral positions”).
Regarding Claim 45, modified Chiu discloses the system of claim 28, and the non-transitory computer-executable instructions (see claim 28 rejection), but does not specifically teach the non-transitory computer-executable instructions which, when executed by the electronic processor, further perform at least: detecting each of the nanoparticles in the population of labeled nanoparticles based on corresponding responses to the illuminating to determine retention times taken for each of the labeled nanoparticles to flow from the first point in or proximal to the fluidic channel to or through the detection zone.
However, Wang, in the same field of flow cytometry, teaches the non-transitory computer-executable instructions which, when executed by the electronic processor (Figure 1; [0029] “a data processing system 22”), further perform at least: detecting each of the nanoparticles in the population of labeled nanoparticles based on corresponding responses to the illuminating (Figure 1; [0029] “a detection system 20 arranged to detect each of the plurality of particles based on corresponding responses to the illumination light”) to determine retention times taken for each of the labeled nanoparticles to flow from the first point in or proximal to the fluidic channel to or through the detection zone (Figure 1; [0029] “determine a time that each of the plurality of particles passes through the detection zone 18”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the system of Chiu with the teachings of Wang, for detecting each of the nanoparticles in the population of labeled nanoparticles based on corresponding responses to the illuminating to determine retention times taken for each of the labeled nanoparticles to flow from the first point in or proximal to the fluidic channel to or through the detection zone, because “CICS has a sheet-like observation volume that enables substantially 100% detection efficiency of single molecules within the separation capillary in contrast to standard laser-induced fluorescence (LIF).” (Wang, [0026])
Regarding Claim 49, modified Chiu discloses the system of claim 28, wherein the non-transitory computer-executable instructions which, when executed by the electronic processor, further perform ([0105] “ranking is performed by a computer and a software representing a ranking algorithm”) at least: distinguishing nanoparticles comprising a payload molecule from nanoparticles lacking a payload molecule in the population of the labeled nanoparticles using at least the signal data ([0202] “A detectable emission of light or any aspect thereof (e.g., an emission peak intensity, an emission intensity range, an emission peak wavelength, an emission wavelength range, an excitation peak wavelength, an excitation wavelength range, an absorption peak wavelength, an absorption wavelength range, an emission lifetime, or a spectral intensity) can be used to determine the presence or absence of a target bionanoparticle. As a result, the methods and systems described herein can be used to facilitate determining, measuring, or indicating the presence or absence of one or more distinct target biological nanoparticles in a fluidic sample, or can be used to facilitate determining or indicating the presence or absence of one or more detectable agents associated with a biological nanoparticle in a fluidic sample”).
Claim 52 is rejected under 35 U.S.C. 103 as being unpatentable over Chiu et al. (US 2021/0016276 A1) and Wang et al. (US 2013/0167623 A1) and Nechev et al. (US 2019/0269795 A1) as applied to claim 28 above, and further in view of Almarsson et al. (US 2018/0085474 A1).
Regarding Claim 52, modified Chiu discloses the system of claim 28, and the non-transitory computer-executable instructions (see claim 28 rejection), but does not specifically teach the non-transitory computer-executable instructions which, when executed by the electronic processor, further perform at least: determining an encapsulation efficiency measure of nanoparticles in the population of labeled nanoparticles.
However, Almarsson, in the same field of lipid nanoparticle compositions, teaches determining an encapsulation efficiency measure of nanoparticles in the population of labeled nanoparticles ([0076] “The encapsulation efficiency may be measured, for example, by comparing the amount of mRNA in a solution containing the nanoparticle composition before and after breaking up the nanoparticle composition with one or more organic solvents or detergents”).
Therefore, it would have been obvious to a person of ordinary skill in the art, before the effective filing date of the claimed invention, to modify the system of Chiu with the teachings of Almarsson, for determining an encapsulation efficiency measure of nanoparticles in the population of labeled nanoparticles, because measuring the encapsulation efficiency of nanoparticles determines the exact amount of a drug, protein, or genetic payload successfully trapped inside the carrier compared to the total amount used, and this metric is vital to ensure therapeutic effectiveness, prevent toxicity from free uncontained agents, and verify manufacturing consistency.
Conclusion
The prior art made of record and not relied upon is considered pertinent to applicant's disclosure.
US-2020/0158615-A1 discloses devices, device systems, and methods for analyzing cells (e.g., blood cells) or particles in a sample. In some embodiments, the disclosure provides various devices and device systems including: a light source; a collecting lens; and one, two, or more detectors. In other embodiments, the devices and device systems include a flow cell or a cartridge device with a flow cell. In further embodiments, the disclosure provides various methods including the steps of: using a light source to emit an irradiation light; using the irradiation light to illuminate a sample flow; using a collecting lens to collect both scattered light and fluorescent light from the sample flow; and using one, two, or more detectors to detect the collected scattered light and fluorescent light. Optionally, these methods include using a flow cell to form a sample flow.
Contact Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Akbar H Rizvi whose telephone number is (571) 272-5085. The examiner can normally be reached Monday - Friday, 9:30 am - 6:30 pm.
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/AKBAR H. RIZVI/
Examiner, Art Unit 2877
/TARIFUR R CHOWDHURY/Supervisory Patent Examiner, Art Unit 2877
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