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 .
Withdrawal of Rejections
The response and amendments filed on 10/21/2025 are acknowledged. Any previously applied minor objections and/or minor rejections (i.e., formal matters), not explicitly restated here for brevity, have been withdrawn necessitated by Applicant’s formality corrections and/or amendments. For the purposes of clarity of the record, the reasons for the Examiner’s withdrawal, and/or maintaining, if applicable, of the substantive or essential claim rejections are detailed directly below and/or in the Examiner’s Response to Arguments section.
Briefly, the previous 35 U.S.C. 112(b) rejections have been withdrawn necessitated by Applicant’s amendments. The previous 35 U.S.C. 103 rejections have been withdrawn necessitated by Applicant’s amendments; however, new grounds of rejection are set forth below.
The following rejections and/or objections are either reiterated or newly applied. They constitute the complete set presently being applied to the instantly application.
Claim Objections
Claim 29 is objected to because of the following informalities: “va lve” should be “valve”. Appropriate correction is required. This is an objection, not a rejection, because this appears to be a typological error.
New Grounds of Rejection Necessitated by Amendments
Claim Rejections - 35 USC § 103, Obviousness
The text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
Claims 10-11, 13, 15-17, 20-22, and 27-28 are rejected under 35 U.S.C. 103 as being unpatentable over Govind (WO 2018/226907; Date of Publication: December 13, 2018 – cited in the IDS filed on 07/14/2023 – previously cited) and Saxl (A fluorescence lifetime-based fibre-optic glucose sensor using glucose/galactose-binding protein; 2011 – cited in the IDS filed on 07/14/2023 – previously cited).
Govind’s general disclosure relates to “a fully integrated microfluidic system capable of producing single-dose amounts of biotherapeutics at the point-of-care wherein protein production, purification and product harvest are all integrated as a single microfluidic device which is portable and capable of continuous-flow production of biotherapeutics at the microscale using a cell-free reaction system” (see, e.g., Govind, abstract). Moreover, Govind discloses that the microfluidic device can capture proteins through substrate binding to binding proteins within chromatography columns (see, e.g., Govind, [0104]), wherein the protein binding can be analyzed using fluorescence (see, e.g., Govind, [0048]).
Regarding claim 10 pertaining to the apparatus, Govind teaches a “factory-on-a-chip microfluidic device”, which can obtain fluid, such as cell-free extracts and reagents, from a sample (see, e.g., Govind, [009], [0015]). Govind teaches microfluidic columns that comprise solid supports comprising immobilized binding protein (see, e.g., Govind, [0051]-[0053]). Govind teaches that the sampling device is connected to the column to allow for protein capture (see, e.g., Govind, [0049]). Govind teaches that the apparatus can comprise a sensor that measures fluorescence (see, e.g., Govind, [0048]). Govind teaches adding the sample to the apparatus, wherein the sample is added to the chromatography column (i.e., microfluidic column) in wash buffer (see, e.g., Govind, [0028]-[0029] & [0104]). Moreover, Govind teaches loading protein, such as GCSF, into the column using a syringe pump to control the flow rate, wherein the flow rate is 0.2 mL/min (see, e.g., Govind, [0104]). Govind teaches that the apparatus can comprise a sensor that measures fluorescence (see, e.g., Govind, [0048]). Additionally, Govind teaches that “devices were stored in a clean and sterile environment until use” (see, e.g., Govind, [0081]).
Regarding claim 11 pertaining to introducing the solid support, Govind teaches introducing a chromatography resin, wherein the resin is preferably an immobilized metal affinity resin and/or an ion exchange resin (see, e.g., Govind, [0014]).
Regarding claim 13 pertaining to the syringe pump, Govind teaches loading protein, such as GCSF, into the column using a syringe pump to control the flow rate, wherein the flow rate is 0.2 mL/min (see, e.g., Govind, [0104]).
Regarding claim 15 pertaining to the flow rate and the amount of substrate binding to binding protein, Govind teaches that “controlled pumping the lysate into the column at an efficiently monitored flow rate would improve the binding” (see, e.g., Govind, [0028]) and “Slightly better controlled pumping of the lysate into the column with a monitored flow rate slightly improved the binding of GFP” (see, e.g., Govind, [0029]). Moreover, Govind teaches equation 5, wherein the diffusion coefficient can be used to solve for the concentration of the particle (see, e.g., Govind, [0098]). Equation 5 also takes into account the concentration of the species (i.e., solute) within the fluid, as well as the fluid velocity (i.e., flow rate) (see, e.g., Govind, [0097]). Additionally, Govind teaches equations 3-4, wherein the diffusion coefficient can be used to solve for the concentration of species in the fluid, wherein the species (i.e., substrate) is available to bind to the binding protein (see, e.g., Govind, [0097]). One of ordinary skill in the art would understand that they can solve the equations set forth by Govind, wherein the flow rate and diffusion can be used to solve for the concentration of substrate that is available to bind to the binding protein. Moreover, one of ordinary skill in the art would readily understand that controlling the flow rate of the substrate, and knowing the starting amount of substrate, would allow for one to determine the amount of substrate that is available to bind to the binding protein, and controlling the flow rate will improve binding, based on the teachings of Govind (see, e.g., Govind, [0028]).
Regarding claim 16 pertaining to controlling the flow rate, Govind teaches controlling the flow rate effectively through the use of a syringe pump, wherein the flow rate is 0.2 mL/min (see, e.g., Govind, [0012], [0104]). One of ordinary skill in the art would recognize that controlling the flow rate would control the amount of protein binding to the binding protein on the chromatography columns, which would control the amount of fluorescence.
Regarding claim 18 pertaining to introducing elution buffer to the system, Govind teaches adding elution buffer to the chromatography columns, wherein the elution buffer is 150 mM imidazole, 1x PBS, pH 7.2 (see, e.g., Govind, [0028]).
Regarding claim 20 pertaining to the method being continuous, Govind teaches that the microfluidic device is capable of continuous flow production (see, e.g., Govind, [008]).
Regarding claim 21 pertaining to the volume of the microfluidic column, Govind teaches that the microfluidic columns can hold a volume ranging from 25-200 µL (see, e.g., Govind, [0014]).
Regarding claim 22 pertaining to the solid support, Govind teaches that the solid support can be “spherical beads with affinity ligands bound to the bead surfaces, wherein the beads are formed of cellulose, poly-styrene-divinylbenzene copolymer, polymethylmethacrylate, or other suitable material” (see, e.g., Govind, [0052]).
Regarding claim 26 pertaining to the apparatus, Govind teaches that the apparatus can contain a syringe pump to control the flow rate (see, e.g., Govind, [0104]).
However, Govind does not teach: wherein the substrate binds to the binding protein and an increase in fluorescence occurs (claim 10, part c); or flowing the binding protein to the solid support for a time necessary to effectuate immobilization of the binding protein to the solid support (claim 11); or wherein the increase in fluorescence is a function of the concentration of substrate bound to the binding protein (claim 17); or wherein the substrate is glucose and the binding protein is a glucose binding protein (claim 27); or wherein the solid support is Ni-NTA agarose beads (claim 28).
Saxl’s general disclosure relates to “fibre-optic biosensors using glucose/galactose binding protein (GBP) labelled with the environmentally sensitive fluorophore, Badan. GBP–Badan was attached via an oligohistidine-tag to the surface of Ni–nitrilotriacetic acid (NTA)-functionalized agarose or polystyrene beads. Fluorescence lifetime increased in response to glucose, observed by fluorescence lifetime imaging microscopy of the GBP–Badan-beads. Either GBP–Badan agarose or polystyrene beads were loaded into a porous chamber at the end of a multimode optical fibre. Fluorescence lifetime responses were recorded using pulsed laser excitation, high speed photodiode detection and time-correlated single photon counting” (see, e.g., Saxl, abstract). Moreover, Saxl discloses that glucose binding to glucose binding protein results in fluorescence as a readout and can be correlated to glucose concentration within a sample (see, e.g., Saxl, Figures 2-3). Furthermore, Saxl teaches that fluorescence-based monitoring of substrate levels “has the advantages of sensitivity and lack of electrochemical interference” (see, e.g., Saxl, Introduction, pg. 968).
Regarding claim 10, part (c) pertaining to the binding of the substrate of the binding protein resulting in increased fluorescence, Saxl teaches that the addition of glucose, and the binding of glucose to glucose binding protein, results in increased fluorescence (see, e.g., Saxl, Figures 2-3). Moreover, Saxl teaches that upon binding to glucose, glucose binding protein undergoes a large change in molecular conformation “which can be detected by either fluorescence resonance energy transfer or by labelling with an environmentally sensitive fluorophore” (see, e.g., Saxl, Introduction, pg. 968).
Regarding claim 11 pertaining to immobilizing the binding protein to the solid support, Saxl teaches that the Ni-NTA agarose beads were incubated with glucose binding protein for four hours (see, e.g., Saxl, “Attachment of GBP-Badan to agarose or polystyrene beads”, pg. 970-971).
Regarding claim 17 pertaining to fluorescence and substrate concentration, Saxl teaches that the concentration of the substrate is positively correlated to fluorescence (see, e.g., Saxl, Figure 2 & Results, pg. 970). Furthermore, increased glucose results in increased binding to glucose binding protein, thereby resulting in increased fluorescence (see, e.g., Saxl, Figures 2-3).
Regarding claim 27 pertaining to the substrate and binding protein, Saxl teaches that the substrate is glucose and the binding protein is glucose binding protein (see, e.g., Saxl, Introduction & Figure 2 & Results, pg. 970).
Regarding claim 28 pertaining to the solid support, Saxl teaches that the solid support is Ni-NTA polystyrene beads (see, e.g., Introduction & Results, pg. 970). Furthermore, Saxl teaches that the sensor’s “response was best conserved using Ni-NTA functionalized agarose beads as an immobilization strategy” and the immobilization of glucose binding protein to Ni-NTA does not impair the fluorescence response to glucose (see, e.g., Saxl, Conclusion, pg. 972).
It would have been first obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce Govind’s microfluidic apparatus, wherein binding of glucose (i.e., substrate) to glucose binding protein (i.e., binding protein) within the chromatography columns would result in increased fluorescence corresponding to the concentration of glucose in the sample, as taught by Saxl. One would have been motivated to do so because Saxl teaches that fluorescence-based monitoring of substrate levels “has the advantages of sensitivity and lack of electrochemical interference” (see, e.g., Saxl, Introduction, pg. 968). Moreover, Saxl teaches that binding of glucose to glucose binding protein results in a large increase in fluorescence intensity, which can be used to determine the concentration of glucose in the sample (see, e.g., Saxl, Introduction, pg. 969 & Figure 2). Additionally, regarding the use of glucose and glucose binding protein, Saxl teaches that upon binding to glucose, glucose binding protein undergoes a large change in molecular conformation “which can be detected by either fluorescence resonance energy transfer or by labelling with an environmentally sensitive fluorophore” (see, e.g., Saxl, Introduction, pg. 968). Furthermore, Govind teaches that the microfluidic columns can be connected to sensors monitoring fluorescence for protein analysis (see, e.g., Govind, [0010]). Therefore, based on the teachings of Govind and Saxl, it would have been obvious to produce Govind’s apparatus, wherein the microfluidic columns are linked to a fluorimeter for measuring the binding of glucose and glucose binding protein in order to determine the concentration of glucose within a sample.
It would have been secondly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce Govind’s microfluidic apparatus, wherein the chromatography columns use Ni-NTA for immobilizing glucose binding protein, as taught by Saxl. One would have been motivated to do so because Saxl teaches that Ni-NTA is used for immobilization of glucose binding protein within the column in order to allow for glucose binding protein to bind to glucose within the sample (see, e.g., Saxl, Introduction, pg. 969 & Results and Discussion, pg. 969). Moreover, Saxl teaches that the sensor’s “response was best conserved using Ni-NTA functionalized agarose beads as an immobilization strategy” and the immobilization of glucose binding protein to Ni-NTA does not impair the fluorescence response to glucose (see, e.g., Saxl, Conclusion, pg. 972). Moreover, Govind teaches that the columns can comprise “immobilized metal affinity resin and/or an ion exchange resin” (see, e.g., Govind, [0014]), wherein Ni-NTA has been used in immobilized metal-affinity chromatography for purification methods (see, e.g., Saxl, Results and Discussion, pg. 969). Therefore, based on the teachings of Govind and Saxl, it would have been obvious to use Ni-NTA within the chromatography columns to immobilize glucose binding protein for binding to glucose within a sample. One would have expected success because Govind and Saxl both teach methods of immobilizing binding proteins to allow for substrate binding and measurement of binding via fluorescence.
Claims 12 and 23 are rejected under 35 U.S.C. 103 as being unpatentable over Govind and Saxl as applied to claims 10-11, 13, 15-17, 20-22, and 27-28 above, and further in view of Ge (On the Possibility of Real-Time Monitoring of Glucose in Cell Culture by Microdialysis Using a Fluorescent Glucose Binding Protein Sensor; 2008 – cited in the IDS filed on 07/14/2023 – previously cited).
The teachings of Govind and Saxl, herein referred to as modified-Govind-Saxl, as discussed above as it pertains to a microfluidic system for measuring the concentration of glucose in a sample.
However, modified-Govind-Saxl does not teach: wherein the sample solution is a culture media solution or a bioreactor solution (claim 12); or wherein the sampling device comprises a microdialysis device (claim 23).
Ge’s general disclosure relates to “glucose sensors with micromolar sensitivity for applications in minimally invasive sampling techniques such as fast microdialysis and extraction of interstitial fluid by iontophoresis and laser poration” (see, e.g., Ge, abstract), wherein the microdialysis device is used to monitor glucose in a solution (see, e.g., Ge, “Experimental Setup”, pg. 692). Moreover, Ge discloses monitoring of glucose consumption in cell culture and fermentation through direct assay of the media in both large and small culture volumes (see, e.g., Ge, Introduction, pg. 692).
Regarding claim 12 pertaining to the sample solution, Ge teaches that the sample solutions are “buffer solutions and mammalian cell culture” (see, e.g., Ge, “Experimental Setup”, pg. 692).
Regarding claim 23 pertaining to a microdialysis device, Ge teaches that a microdialysis device to monitor glucose in a solution, such as buffer solutions and mammalian cell culture (see, e.g., Ge, “Experimental Setup”, pg. 692). Furthermore, Ge teaches that the use of a microdialysis device allows for monitoring of glucose levels in small culture volumes (see, e.g., Ge, Introduction, pg. 692).
It would have been first obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to employ modified-Govind-Saxl’s microfluidic apparatus to measure substrate concentrations in buffer solutions and mammalian cell culture, as taught by Ge. One would have been motivated to do so because Ge teaches that measuring glucose levels allows for real-time monitoring of glucose levels within the culture, which can be correlated to cell growth, because glucose levels decrease as cell grow and consume glucose (see, e.g., “Mammalian Cell Culture”, pg. 696 & Figure 8). Moreover, modified-Govind-Saxl teaches that crude protein can be captured within a sample by the microfluidic device and sensors can be employed for monitoring various aspects of the protein (see, e.g., Govind, [0010]), such as protein concentration using fluorescence (see, e.g., Saxl, Figure 2 & Results, pg. 970). Moreover, modified-Govind-Saxl teaches capturing, eluting, and providing concentrations of G-CSF (see, e.g., Govind, [0035]), which one of ordinary skill in the art would understand can be applied to various samples, such as cell culture samples with glucose as the substrate. Therefore, based on the teachings of modified-Govind-Saxl and Ge, it would have been obvious to measure glucose concentrations in cell culture using a microfluidic device because this would allow for one of ordinary skill in the art to measure glucose concentrations and glucose consumption by cells and correlate it to cell growth.
It would have been secondly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce modified-Govind-Saxl’s microfluidic apparatus, wherein the microfluidic apparatus contains a microdialysis device as the sampling device, as taught by Ge. One would have been motivated to do so because Ge teaches that the use of a microdialysis device allows for monitoring of glucose levels in small culture volumes (see, e.g., Ge, Introduction, pg. 692). Moreover, Ge teaches that physiological concentrations of glucose, which are in the millimolar range, require a dilution step prior to measurement; however, sampling techniques such as microdialysis inherently dilutes the sample (see, e.g., Ge, Results and Discussion, pg. 693). Furthermore, modified-Govind-Saxl teaches the use of dialysis cassettes and bags for collecting protein within a sample (see, e.g., Govind, [00101]-[00103]), and dilution of the sample before immobilization via the chromatography columns (see, e.g., Govind, [0025]). Therefore, based on the teachings of modified-Govind-Saxl and Ge, it would have been obvious to produce a microfluidic device, wherein the sampling device is a microdialysis device used to collect/sample small amounts of glucose in a sample for further downstream analysis, such as measuring protein concentration. One would have expected success because modified-Govind-Saxl and Ge both teach the dialysis techniques for protein sampling.
Claims 14 and 24-25 are rejected under 35 U.S.C. 103 as being unpatentable over Govind and Saxl as applied to claims 10-11, 13, 15-17, 20-22, and 27-28 above, and further in view of Kostov (Portable system for the detection of micromolar concentrations of glucose; 2014 – cited in the IDS filed on 07/14/2023 – previously cited).
The teachings of Govind and Saxl, herein referred to as modified-Govind-Saxl, as discussed above as it pertains to a microfluidic system for measuring the concentration of glucose in a sample.
However, modified-Govind-Saxl does not teach: determining the concentration of the concentration of the substrate in the sample solution using the measured fluorescence and a calibration curve (claim 14); or wherein the fluorometer is a microfluorometer (claim 24); or wherein the microfluidic column is positions flush with the fluorimeter to facilitate fluorescent measurements (claim 25).
Kostov’s general disclosure relates to “a USB powered, all-solid-state, hand-held fluorometric system for measurement of low glucose concentrations. It utilizes the dual-labeled GBP as sensor and a dedicated low cost, dual emission microfluorometer” (see, e.g., Kostov, Introduction, pg. 2). Moreover, Kostov discloses the construction of calibration curves in order to measure the concentration of substrate within a sample, such as the amount of glucose within a biological sample (see, e.g., Kostov, Results and Discussion, pgs. 7-9 & Figures 5-6).
Regarding claim 14 pertaining to determine the concentration of the substrate, Kostov teaches constructing a calibration curve (see, e.g., Kostov, “Calibration”, pg. 6) and using the measured fluorescence to determine the concentration of the substrate within the sample (see, e.g., Kostov, Results and Discussion, pgs. 7-9 & Figures 5-6).
Regarding claim 24 pertaining to the microfluorometer, Kostov teaches using a dual emission microfluorometer (see, e.g., Kostov, Introduction, pg. 2). Moreover, Kostov teaches that using a microfluorometer system is a promising tool for accurate measurements of low glucose concentrations in biological samples (see, e.g., Kostov, Conclusion, pg. 9).
Regarding claim 25 pertaining to the column being flush with the fluorimeter, Kostov teaches that the fluorometric system contains a cuvette, which holds the sample, and a microfluorometer, wherein the cuvette holding the sample is sandwiched between the photodiodes of the microfluorometer (see, e.g., Kostov, “Optics”, pg. 5 & Figure 2).
It would have been first obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to employ modified-Govind-Saxl’s microfluidic device in order to determine the concentration of a substrate by constructing a calibration curve, as taught by Kostov. One would have been motivated to do so because Kostov teaches that the calibration curve can be constructed to determine the amount of binding between glucose and glucose binding protein, as well the concentration of glucose in a sample via fluorescent measurements (see, e.g., Kostov, “Calibration”, pg. 6). Moreover, Kostov teaches that over-labelling or under-labelling the molecules will affect the calibration curve and the fluorescence readouts (see, e.g., Kostov, “Fluorometer design and operation”, pg. 8); therefore, one of ordinary skill in the art would understand that constructing the calibration curve would also ensure that molecules are labelled correctly. Furthermore, modified-Govind-Saxl teaches the measurement of glucose concentrations by via fluorescent read-outs of glucose binding to glucose binding protein within a microfluidic device (see, e.g., Saxl, Figures 2-3) (see, e.g., Govind, [009], [0015]). Therefore, based on the teachings of modified-Govind-Saxl and Kostov, it would have been obvious to construct a calibration curve when measuring glucose binding to glucose binding protein because this would allow for one to accurately calculate the concentration of glucose in a sample by measuring fluorescence.
It would have been secondly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce modified-Govind-Saxl’s microfluidic device, wherein the microfluidic device contains a microfluorometer, as taught by Kostov. One would have been motivated to do so because Kostov teaches that using a microfluorometer system is a promising tool for accurate measurements of low glucose concentrations in biological samples (see, e.g., Kostov, Conclusion, pg. 9). Moreover, Kostov teaches that using a microfluorometer allows for the production of a handheld glucose detection system that can measure the amount of glucose in a sample using fluorescence via glucose binding to glucose binding protein (see, e.g., Kostov, “Theory of Operation”, pgs. 2-3 & “Microfluorometer system”, pg. 4). Furthermore, modified-Govind-Saxl teaches the measurement of glucose concentrations by via fluorescent read-outs of glucose binding to glucose binding protein within a microfluidic device (see, e.g., Saxl, Figures 2-3) (see, e.g., Govind, [009], [0015]). Therefore, based on the teachings of modified-Govind-Saxl and Kostov, it would have been obvious to construct a microfluidic device to measure glucose concentrations in a sample, wherein the microfluidic device contains a microfluorometer because the microfluorometer allows for detection of low glucose concentrations in a biological sample via fluorescent readouts resulting from glucose binding to glucose binding protein, and allows for the device to be handheld.
It would have been thirdly obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce modified-Govind-Saxl’s microfluidic device, wherein the microfluidic device requires the column to be flush with the microfluorometer, as taught by Kostov. One would have been motivated to do so because Kostov teaches that the labelled glucose binding protein can be excited at a specific wavelength, and the emission can be detected at a specific wavelength (see, e.g., Kostov, “Optics”, pgs. 4-5); therefore, “sandwiching” the cuvette between photodiodes allows for the photodiodes to select for the emission wavelengths that are emitted from excitation of the glucose binding protein within the sample (see, e.g., Kostov, “Optics”, pgs. 4-5); thereby acting as a detector for the microfluorometer (see, e.g., Kostov, “Fluorometer design and operation”, pg. 7). Additionally, based on the teachings of Kostov’s handheld glucose monitoring device (see, e.g., Kostov, “Theory of Operation”, pgs. 2-3 & “Microfluorometer system”, pg. 4), one of ordinary skill in the art would understand that placing the microfluidic device flush with the microfluorometer would allow for the production of a smaller device for measuring glucose concentrations in a sample. Furthermore, modified-Govind-Saxl teaches the measurement of glucose concentrations by via fluorescent read-outs of glucose binding to glucose binding protein within a microfluidic device, also known as a factory-on-a-chip microfluidic device, which is small in scale (see, e.g., Saxl, Figures 2-3) (see, e.g., Govind, [009], [0015]). Therefore, based on the teachings of modified-Govind-Saxl and Kostov, it would have been obvious to produce a microfluidic device for measuring glucose concentrations, wherein the microfluidic device is flush with the microfluorometer because this allows for the photodiodes within the microfluorometer to detect the emission wavelengths emitted from the glucose binding proteins. One would have expected success because modified-Govind-Saxl and Kostov both teach measuring glucose concentrations in a sample using fluorescence.
Claims 29-31 are rejected under 35 U.S.C. 103 as being unpatentable over Govind and Saxl as applied to claims 10-11, 13, 15-17, 20-22, and 27-28 above, and further in view of Gamble (U.S. Patent Number 7,178,386; Date of Publication: February 20, 2007 – newly cited).
The teachings of Govind and Saxl, herein referred to as modified-Govind-Saxl, as discussed above as it pertains to a microfluidic system for measuring the concentration of glucose in a sample.
However, modified-Govind-Saxl does not teach: collecting the substrate-containing buffer in a sample tube prior to introduction to the microfluidic column, wherein the sample tube is positioned upstream of the microfluidic column with a first valve positioned therebetween, wherein during collection of the substrate-containing buffer in the sample tube, the first valve is in a first position, prohibiting entry of the substrate-containing buffer to the microfluidic column (claim 29); or introducing the substrate-containing buffer collected in the sample tube to the microfluidic column for fluorescence measurement, said introduction comprising positioning the first valve in a second position and directing additional buffer to the sample tube instead of the aseptic sampling device using one or more additional valves (claim 30); or wherein subsequent to the fluorescence measurement, continued introduction of buffer to the sample tube and the microfluidic device elutes the substrate from the binding protein (claim 31).
Gamble’s general disclosure relates to “A parallel fluid processing system including multiple fluid process regions containing solid material in fluid communication with a common first fluid source may be used to conduct analyses and/or synthesis in parallel. A parallel fluid processing data correction method includes supplying and processing a calibrant in each fluid process region, measuring a first physical parameter and deriving at least one correction factor based on the parameter, supplying and processing at least one second fluid in each fluid process region, and then applying the correction factor to yield corrected process data. Retention time correction, peak area correction, and other useful data corrections may be performed. Parallel fluid processing may be performed with microfluidic devices and systems” (see, e.g., Gamble, abstract).
Regarding claims 29-31 pertaining to the sample tube, valves, and microfluidic column, Gamble teaches a sample source (see, e.g., Gamble, Figure 2, Sample Source 40), which one of ordinary skill in the art would recognize can be a sample tube, that is positioned upstream of a column with a first rotary valve positioned in between (see, e.g., Gamble, Figure 2). Additionally, Gamble teaches that the rotary valve can be controlled to inject a predetermined about of sample from the sample source into the column (see, e.g., Gamble, pg. 31, col. 3, lines 37-40). One of ordinary skill in the art would readily understand that if the valve can inject a predetermined amount of sample, that the valve can be opened and closed in order to prohibit entry or allow entry of the substrate-containing buffer into the column. Furthermore, Gamble teaches a waste reservoir 44 for substrate that has been eluted from the binding protein within the column (see, e.g., Gamble, Figure 2). Gamble teaches a high pressure pump 34 within the device to pressurize mobile phase solvent from the reservoir so that when the sample source and the solvent interact at injection valve 38, the mixture is sent through column 10 and the mixture is eluted from the detector 42 to the waste reservoir 44 (see, e.g., Gamble, Figure 2). Gamble teaches that system 30 is often operated on a nearly continuous basis (see, e.g., Gamble, pg. 31, col. 3, lines 52-53 & pg. 35, col. 11, lines 9-15).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to produce a microfluidic system for measuring the concentration of glucose in a sample, as taught by modified-Govind-Saxl, wherein the system has a sample tube located upstream of the microfluidic column and wherein the sample is introduced to the column by a valve and subsequently eluted, as taught by Gamble. One would have been motivated to do so because Gamble teaches that by introducing a sample upstream of the microfluidic column, wherein the sample has to pass through a rotary valve, that the rotary valve can be controlled to inject a predetermined about of sample from the sample source into the column (see, e.g., Gamble, pg. 31, col. 3, lines 37-40), wherein one of ordinary skill in the art would understand that the valve can be opened and closed to allow or prohibit entry of the sample into the column for detection. Moreover, modified-Govind-Saxl teaches adding a sample to an apparatus, wherein the sample is added to a chromatography column (i.e., microfluidic column) in wash buffer (see, e.g., Govind, [0028]-[0029] & [0104]). Moreover, modified-Govind-Saxl teaches loading protein, such as GCSF, into the column using a syringe pump to control the flow rate, wherein the flow rate is 0.2 mL/min (see, e.g., Govind, [0104]). Additionally, modified-Govind-Saxl teaches “controlled pumping the lysate into the column at an efficiently monitored flow rate would improve the binding” (see, e.g., Govind, [0028]) and elution of the protein from the microfluidic chromatography column (see, e.g., Govind, [0028]-[0029]). Therefore, based on the teachings of modified-Govind-Saxl and Gamble, it would have been obvious to combine the teachings and apparatuses taught by modified-Govind-Saxl and Gamble because injecting a sample through a valve before introduction into the microfluidic column allows for a pre-measured amount of sample, as a specific flow rate, to be introduced into the column for detection of fluorescence.
Examiner’s Response to Arguments
Applicant’s arguments filed on 10/21/2025 have been fully considered but they are not persuasive.
Regarding Applicant’s arguments pertaining to the sampling device and Govind’s teachings pertaining to the mixer/de-bubbler unit (remarks, pages 6-7), these arguments are not persuasive because nowhere in the Examiner’s non-final office action did the Examiner state that the mixer/de-bubbler unit taught by Govind is the claimed sampling device. Instead, the Examiner was reiterating the teachings of Govind which teach “Microfluidic purification unit (120) in Figure 1 is communicatively connected to the microfluidic mixer/de-bubbler unit mixer device and contain a modular chip based purification column or columns for protein capture, buffer-exchange and polishing the protein harvest” (see, e.g., Govind, [0050]). Therefore, these teachings show that the microfluidic device contains a mixer/de-bubbler unit (see, e.g., Govind, [0049]), and a column for protein capture, but the Examiner did not mean to convey that the mixer/de-bubbler unit is the sampling device. Despite this, Govind does indeed teach a sampling device, wherein the device captures proteins from a sample on a purification column (see, e.g., Govind, Figure 1).
Regarding Applicant’s arguments pertaining to the sampling device being inserted into the sample solution (remarks, pages 6-7), these arguments are not persuasive because Saxl teaches inserting a fiber-optic glucose sensor into a sample solution in order to measure the amount of glucose within a sample through interaction of the glucose with the GBP-Badan labelled beads (see, e.g., Saxl, Figure 5 & “Glucose sensor responses”, pg. 971).
Regarding Applicant’s arguments pertaining to substrate-containing buffer (remarks, pages 7-8), these arguments are not persuasive because the broadest reasonable interpretation (BRI) of independent claim 10(b) pertains to a substrate diffusing from a sample solution into a buffer when mixed together. Govind teaches a microfluidic device, wherein a sample can be mixed and diluted with a buffer before microfluidic purification (see, e.g., Govind, [0010], Figure 7). Furthermore, it is unclear from Applicant’s instant Specification if there is 0% sample solution that mixes or diffuses in the buffer when the substrate diffuses from the sample solution to the buffer to form the substrate-containing buffer. In other words, there is no evidence that 100% of the substrate diffuses from the sample solution into the buffer without any residual mixture of the sample solution into the buffer at the same time. Without this evidence, the skilled artisan is unable to discern if there is crude material within the microfluidic bioreactor. Moreover, if there is sample solution, at any amount, that mixes with the buffer solution, then Applicant is merely claiming a step of mixing the sample solution and buffer together. MPEP716.01(c)(II) states that “Arguments presented by the applicant cannot take the place of evidence in the record. In re Schulze, 346 F.2d 600, 602, 145 USPQ 716,718 (CCPA 1965) and In re De Blauwe, 736 F.2d 699, 705, 222 USPQ 191, 196 (Fed. Cir.1984)”; therefore, Applicant’s arguments are not sufficient to overcome the prior arts of record because Applicant has not provided objective evidence.
In response to applicant’s argument that there is no teaching, suggestion, or motivation to combine the references of Govind and Ge (remarks, page 9), the examiner recognizes that obviousness may be established by combining or modifying the teachings of the prior art to produce the claimed invention where there is some teaching, suggestion, or motivation to do so found either in the references themselves or in the knowledge generally available to one of ordinary skill in the art. See In re Fine, 837 F.2d 1071, 5 USPQ2d 1596 (Fed. Cir. 1988), In re Jones, 958 F.2d 347, 21 USPQ2d 1941 (Fed. Cir. 1992), and KSR International Co. v. Teleflex, Inc., 550 U.S. 398, 82 USPQ2d 1385 (2007). In this case, both modified-Govind-Saxl and Ge’s teachings pertain to dialysis techniques for protein sampling. Moreover, Ge teaches real-time monitoring of glucose samples through fluorescent measurements (see, e.g., “Mammalian Cell Culture”, pg. 696 & Figure 8), which can be applied to the method(s) and device(s) taught by modified-Govind-Saxl. Therefore, based on the teachings of modified-Govind-Saxl and Ge there is motivation to combine the teachings.
Conclusion
Claims 10-17, 20-25, and 27-31 are rejected.
No claims are allowed.
THIS ACTION IS MADE FINAL. Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
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/NATALIE IANNUZO/Examiner, Art Unit 1653
/SHARMILA G LANDAU/Supervisory Patent Examiner, Art Unit 1653
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