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 .
Remarks
This Office Action fully acknowledges Applicant’s remarks filed on 01/15/2026. Claims 1-6, 8, 10, 12, 13, 16, 17, 19-22, 24, 28, 30, 32, 34-36 are pending. Claims 34-36 are newly added.
Drawings
The drawings are objected to under 37 CFR 1.83(a). The drawings must show every feature of the invention specified in the claims. Therefore, the flow cell recited in claims 1and 21 must be shown or the feature(s) canceled from the claim(s). No new matter should be entered.
Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. The figure or figure number of an amended drawing should not be labeled as “amended.” If a drawing figure is to be canceled, the appropriate figure must be removed from the replacement sheet, and where necessary, the remaining figures must be renumbered and appropriate changes made to the brief description of the several views of the drawings for consistency. Additional replacement sheets may be necessary to show the renumbering of the remaining figures. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance.
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
1. Claims 1, 5, 6, 12, 13 and 20 are rejected under 35 U.S.C. 103 as being unpatentable over U.S. Patent Application Publication No. 2016/0266087 to Lo et al. (cited by applicant) in view of Irish Patent Application Publication No. IE20060796 to Kelleher et al. (cited by applicant)
Lo et al. discloses devices and methods for differentiating proteins by frequency domain fluorescence lifetime spectroscopy that provides for quantifying the concentration of a protein in a mixture ([0012], [0054]), wherein the protein of interest has an intrinsic fluorescence decay signature ([0042])
The method of Lo et al. includes directing light from a modulated light source onto a mixture of proteins, wherein the light has a wavelength of about 300 nm ([0048]), and taking a series of measurements of the fluorescence intensity of the mixture at a series of time points; wherein the phase shift between the modulated light source and the modulated fluorescent light is recorded and wherein decay times which differ from each other by less than a nanosecond up to 10 ns are deducible from the phase shift measurements ([0062], [0065]-[0073], [0084], [0086]), and quantifying the concentration of the protein of interest in the sample by reference to the fluorescence decay model (curve fit) ([0066]).
Lo et al. teaches excitation light in a wavelength of 300 nm and above, but does not teach using excitation light in a wavelength of 240-295 nm.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to conduct routine engineering optimization experimentation to determine excitation wavelengths to use for different proteins including wavelengths of 240 to 295 nm.
Lo et al. discuss sub-nanosecond excitation light pulses in paragraph [0042].
In paragraph [0042] Lo et al. also discusses how the time domain method provides significant information over shorter pulses. Lo et al. does not teach the specific time intervals recited in claim 1.
Kelleher et al. teaches using fluorescent excitation pulses that are less than a nanosecond (480 picoseconds) to excite particles to be analyzed using decay times of the emission. (page 18, second paragraph and Figs. 15(a) and 15 (b)).
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing dates to modify Lo et al. to use pulses of excitation light and measurements of fluorescent emission within a time frame of less than a nanosecond as taught by Kelleher et al. It would further have been obvious to for one skilled in the art to conduct routine engineering optimization experimentation to provide and record a series of time intervals between fluorescence measurements and preceding light pulses that differ from each other by less than a nanosecond as taught by Kelleher et al. and provide differences between largest and smallest time intervals that is at least 10 ns and/or a sufficient time to detect a decay of the fluorescence intensity towards a baseline level.
As to the newly added limitation that the mixture is introduced into a flow cell wherein the mixture is exposed to one or more pulses of light, Kelleher et al. teaches that detection can be accomplished using flow cytometry which renders it obvious to use a flow cell to differentiate proteins in a mixture that is passed through a flow cell and irritated with fluorescent radiation.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to differentiate proteins in mixtures via a flow cells, based on Kelleher et al. teaching flow cytometry for such analysis.
I.) As noted above, Lo et al. in view of Kelleher et al. renders all the limitations of applicant’s claim 1 obvious.
Therefore, Lo et al. in view of Kelleher et al. renders claim 1 obvious.
II.) Regarding applicant’s claim 5, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 5 depends.
Claim 5 recites that the mixture comprises more than one protein, which each have a different intrinsic fluorescence decay signature.
Lo et al. teaches different proteins that have different intrinsic fluorescence decay. [0012]
Therefore, Lo et al. in view of Kelleher et al. renders claim 5 obvious.
III.) Regarding applicant’s claim 6, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 6 depends.
Claim 6 recites that the fluorescence decay curve is measured multiple times, to allow a change in the concentration of the protein of interest over time and/or to determine the concentration of the protein of interest in more than one eluate fraction to be determined.
Lo et al. in view of Kelleher et al. does not teach that the fluorescence decay curve is measured multiple times, to allow a change in the concentration of the protein of interest over time.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to measure the fluorescence decay curve multiple times to detect any change in the concentration of the protein of interest over time.
Therefore, Lo et al. in view of Kelleher et al. renders claim 6 obvious.
IV.) Regarding applicant’s claim 12, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 12 depends.
Claim 12 recites that the one or more pulses of light have a pulse width is less than 10 ns.
Lo et al. in view of Kelleher et al. does not teach that the one or more pulses of light have a pulse width is less than 10 ns.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to conduct routine engineering optimization experimentation to determine light pulse widths that are appropriate for proteins of interest to be quantified, including pulses of light having a pulse width is less than 10 ns.
V.) Regarding applicant’s claim 13, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 13 depends.
Claim 13 recites that wherein the fluorescence decay curve is fitted to a single-exponential or double-exponential model.
Kelleher et al. teaches three exponential analysis of decay curves in Figs. 4, 5 and 7.
In Lo et al. in view Kelleher et al. it would have been obvious to fit the fluorescence decay curve using a double-exponential model.
Therefore, Lo et al. in view of Kelleher et al. renders claim 13 obvious.
VI.) Regarding applicant’s claim 20, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 20 depends.
Claim 20 recites each protein species has a characteristic ƬI, Ƭ2 and β that can be used to identify that species, where in a sum of two exponential decays model, ƬI and Ƭ2 are the first and second fluorescence decay times and β is the contribution of the first decay component.
Since each protein species has unique characteristics which would inherently contribute to fluorescent decay, it would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to use the unique characteristics of the individual proteins to identify the individual proteins.
Therefore, Lo et al. in view of Kelleher et al. renders claim 20 obvious.
VI.) Regarding applicant’s claim 34, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 34 depends.
Claim 34 recites that the light has a wavelength in the 250-280 nm range, optionally wherein the light has a wavelength of 266 nm.
Lo et al. in view of Kelleher et al. does not teach that the light has a wavelength in the range 250-280 nm range, optionally wherein the light has a wavelength of 266 nm.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to conduct routine engineering optimization experimentation to determine excitation wavelengths to use for different proteins including wavelengths of 240 to 295 nm.
Therefore, Lo et al. in view of Kelleher et al. renders claim 34 obvious.
VII.) Regarding applicant’s claim 35, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 35 depends.
Claim 35 recites that the one or more pulses of light have a pulse width which is less than 5 ns, optionally less than 2 ns.
Lo et al. in view of Kelleher et al. does not teach that the light has one or more pulses of light have a pulse width which is less than 5 ns, optionally less than 2 ns.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to conduct routine engineering optimization experimentation to determine suitable pulse widths to use for different proteins including a pulse width which is less than 5 ns, optionally less than 2 ns.
Therefore, Lo et al. in view of Kelleher et al. renders claim 35 obvious.
2. Claims 2, 3, 8 and 10 are rejected under 35 U.S.C. 103 as being unpatentable over Lo et al. in view of Kelleher et al. as applied to claims 1 and 5 above, and further in view of U.S. Patent Application Publication No. 2009/0316992 to Wheelock.
I.) Regarding applicant’s claim 2, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 2 depends.
Claim 2 recites that the quantification of the concentration of the protein of interest or the concentration of the conformational state of the protein of interest comprises deconvoluting the fluorescence decay curve to quantify the contribution of the intrinsic fluorescence decay signature from different proteins species or conformational states of the protein species to the fluorescence decay curve.
Kelleher et al. teaches that reconvolution (deconvolution) of peak decay data is a standard procedure is pulsed fluorescence decay measurements. (page 25, lines 14-24)
Lo et al. in view of Kelleher et al. does not teach quantification of the concentration of the protein of interest or the concentration of the conformational state of the protein of interest by deconvoluting the fluorescence decay curve to quantify the contribution of the intrinsic fluorescence decay signature from different proteins species or conformational states of the protein species to the fluorescence decay curve.
Wheelock teaches time-resolved emission of proteins that utilizes differences in fluorescent lifetime imaging to differentiate between fluorescence from specific protein labels and non-specific background fluorescence.
Wheelock teaches the use of deconvolution algorithms for multi-exponential decay, ([0044]) and using the entire area under the curve (AUC) for quantification purposes the AUC, corresponding to the fitted photon count for each specific component. [0051]
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to use deconvolution of analysis the fluorescence decay curve to quantify the contribution of the intrinsic fluorescence decay signature from different proteins species or conformational states of the protein species to the fluorescence decay curve as taught by Wheelock.
Therefore, Lo et al. in view of Kelleher et al. and Wheelock renders claim 2 obvious.
II.) Regarding applicant’s claim 3, as noted above Lo et al. in view of Kelleher et al. and Wheelock renders claim 2 obvious from which claim 3 depends.
Claim 3 recites that the quantification of the concentration of the protein of interest or the concentration of the conformational state of the protein of interest comprises calculating the area under the deconvoluted portion of the fluorescence decay curve that corresponds to the intrinsic fluorescence decay signature of the protein of interest or of a conformational state of the protein of interest.
As noted above, Wheelock teaches using the entire area under the curve (AUC) for quantification purposes the AUC, corresponding to the fitted photon count for each specific component. [0051]
Kelleher et al. teaches analysis of dependence of integrated photoluminescence (PL) intensity decays and values of averaged lifetimes on different concentrations of BSA protein, as shown in bar charts of Figs. 8-11 which are based on the areas of the deconvoluted peaks. (page 26, lines 7-10 and 15-20)
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. and Wheelock to calculate the area under the deconvoluted portion of the fluorescence decay curves that corresponds to the intrinsic fluorescence decay signature of the protein of interest or of a conformational state of the protein of interest for purposes of quantification of the concentration of the protein of interest or the concentration of the conformational state of the protein of interest as taught by Wheelock and Kelleher et al.
Therefore, Lo et al, in view of Kelleher et al. and Wheelock renders claim 3 obvious.
III.) Regarding applicant’s claim 8, as noted above Lo et al. in view of Kelleher et al. renders claim 5 obvious from which claim 8 depends.
Claim 8 recites that the concentration of more than one protein is quantified, and wherein the concentrations of the proteins are quantified by deconvoluting more than one intrinsic fluorescence decay signature from a single fluorescence decay curve.
As noted above, Kelleher et al. teaches that reconvolution (deconvolution) of peak decay data is a standard procedure is pulsed fluorescence decay measurements. (page 25, lines 14-24)
Lo et al. in view of Kelleher et al. does not teach quantification of the concentration of the protein of interest or the concentration of the conformational state of the protein of interest by deconvoluting the fluorescence decay curve to quantify the contribution of the intrinsic fluorescence decay signature from different proteins species or conformational states of the protein species to the fluorescence decay curve.
Wheelock teaches time-resolved emission of proteins that utilizes differences in fluorescent lifetime imaging to differentiate between fluorescence from specific protein labels and non-specific background fluorescence.
Wheelock teaches the use of deconvolution algorithms for multi-exponential decay, ([0044]) and using the entire area under the curve (AUC) for quantification purposes the AUC, corresponding to the fitted photon count for each specific component. [0051]
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to use deconvolution of analysis the fluorescence decay curve to quantify the contribution of the intrinsic fluorescence decay signature from different proteins species or conformational states of the protein species to the fluorescence decay curve as taught by Wheelock.
It would have further been obvious to quantify the concentration of more than one protein by deconvoluting more than one intrinsic fluorescence decay signature from a single fluorescence decay curve that includes fluorescence decay signature from a different proteins for purposes of quantifying different proteins in a mixture.
Therefore, Lo et al. in view of Kelleher et al. and Wheelock renders claim 8 obvious.
IV.) Regarding applicant’s claim 10, as noted above Lo et al. in view of Kelleher et al. renders claim 8 obvious from which claim 10 depends.
Claim 10 recites that the concentrations of the proteins are quantified by deconvoluting a first intrinsic fluorescence decay signature from a first mixture and a second intrinsic fluorescence decay signature from a second mixture, wherein the first and the second mixtures are eluate from a column at different elution times; or by deconvoluting a first intrinsic fluorescence decay signature and a second intrinsic fluorescence decay signature from a single mixture.
As noted above, it would have further been obvious to quantify the concentration of more than one protein by deconvoluting more than one intrinsic fluorescence decay signature from a single fluorescence decay curve.
Further, since different proteins have different intrinsic decay signatures, it would have been obvious to one of ordinary skill in the art to quantify concentrations of different proteins by deconvoluting a first intrinsic fluorescence decay signature and a second intrinsic fluorescence decay signature from a single mixture.
Therefore, Lo et al. in view if Kelleher et al. and Wheelock renders claim 10 obvious.
3. Claim 4, 17 and 19 are rejected under 35 U.S.C. 103 as being unpatentable over Lo et al. in view of Kelleher et al. as applied to claim 1 above, and further in view of Applicant’s Admitted Prior Art.
I.) Regarding applicant’s claim 4, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 4 depends.
Claim 4 recites that the mixture is a portion of an eluate from a chromatography column.
Lo et al. in view of Kelleher et al. does not teach that the mixture is a portion of an eluate from a chromatography column.
In the Background section of the disclosure applicant notes that chromatography is commonly used is protein separation for bioprocessing. On page 2, lines 1-2 applicant nots that “In a typical industrial bioprocess, chromatography is the key step for purifying the product from other proteins, cellular debris and components of the media.”
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to receive and quantify proteins from the eluate of a chromatography column as taught by Applicant’s Admitted Prior Art for purposes of bioprocessing.
Therefore, Lo et al. in view of Kelleher et al. and Applicant’s Admitted Prior Art renders claim 4 obvious.
II.) Regarding applicant’s claim 17, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 17 depends.
Claim 17 recites the method further comprises a generation of one or more decay chromatograms, DCs, by fitting a double, DC-2, exponential model to the fluorescence decay curve; and/or wherein the method further comprises a generation of one or more decay-associated chromatograms, DACs, by fitting equation (8) to the fluorescence decay curve, and calculating the contribution of each proteins species to the fluorescence intensity measured across the time window using equation (14).
Lo et al. in view of Kelleher et al does not teach generation of one or more decay chromatograms by fitting a double exponential model to the fluorescence decay curve; and/or wherein the method further comprises a generation of one or more decay-associated chromatograms and calculating the contribution of each proteins species to the fluorescence intensity measured across the time window of the chromatograms for quantifying proteins in the eluate of the chromatography column.
As noted above, it would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to receive and quantify proteins from the eluate of a chromatography column as taught by Applicant’s Admitted Prior Art for purposes of bioprocessing.
Chromatograms are time-based graphic records (as of concentration of eluted materials) of a chromatographic separation (https://www.merriam-webster.com/dictionary/chromatogram)
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to generation of one or more decay chromatograms by fitting a double exponential model to the fluorescence decay curve; and/or wherein the method further comprises a generation of one or more decay-associated chromatograms and calculating the contribution of each proteins species to the fluorescence intensity measured across the time window of the chromatograms for quantifying proteins in the eluate of the chromatography column.
One skilled in the art would find it obvious to use the variables in the formula recited in claim 17 to generate chromatograms using the same formula or an equivalent formula to generate the chromatograms.
Therefore, Lo et al. in view of Kelleher et al. and Applicant’s Admitted Prior Art renders claim 17 obvious.
III.) Regarding applicant’s claim 19, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 19 depends.
Claim 19 recites that the method further comprises quantifying two co-eluting proteins by simultaneous measurement of both a time decay and the fluorescence intensity.
Lo et al. in view of Kelleher et al. does not teach quantifying two co-eluting proteins by simultaneous measurement of both a time decay and the fluorescence intensity.
As noted above, it would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to receive and quantify proteins from the eluate of a chromatography column as taught by Applicant’s Admitted Prior Art for purposes of bioprocessing.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to teach quantify two co-eluting proteins by simultaneous measurement of both a time decay and the fluorescence intensity, for purposes of quantifying proteins in the eluate of a chromatography column.
Therefore Lo et al. in view of Kelleher et al. and Applicant’s Admitted Prior Art renders claim 19 obvious.
4. Claim 16 is rejected under 35 U.S.C. 103 as being unpatentable over Lo et al. in view of Kelleher et al. as applied to claim 1 above, and further in view of U.S. Patent No. 3,502,993 to Schurzinger et al.
I.) Regarding applicant’s claim 16, as noted above Lo et al. in view of Kelleher et al. renders claim 1 obvious from which claim 16 depends.
Claim 16 recites that contribution of a background noise signal, Ibackground(t), is calculated using the following equation: Ibackground(t) = cƟ where c is a baseline offset value and Ɵ is a width of the time window of a high-bandwidth digitizer or a sampling oscilloscope.
Lo et al. in view of Kelleher et al. does not teach calculating background noise signal by multiplying a baseline offset by the width of the time window of a high-bandwidth digitizer.
Schurzinger et al. teaches digitizing signals and compensating for changes in background levels by monitoring amplitude variations.
It would have been obvious to one of ordinary skilled in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to digitize signals and determine background levels by monitoring amplitude (width) variations.
Therefore, Lo et al. in view of Kelleher et al. and Schurzinger et al. renders claim 16 obvious.
5. Claims 21, 22, 24 and 30 are rejected under 35 U.S.C. 103 as being unpatentable over Lo et al. in view of Kelleher et al.
Lo et al. discloses devices and methods for differentiating proteins by frequency domain fluorescence lifetime spectroscopy that provides for quantifying the concentration of a protein of interest or the concentration of a conformational state of the protein of interest, in a mixture ([0012], [0054]), wherein the protein of interest or conformational state has an intrinsic fluorescence decay signature ([0042])
The apparatus of Lo et al. includes a modulated light source, a focusing optical fiber, a detecting optical fiber, and a detector. (Abstract). Lo et al. discloses a laser light source. [0006]
The method of Lo et al. includes directing light from a modulated laser light source onto a mixture of proteins, wherein the light has a wavelength of about 300 nm ([0048]), and taking a series of measurements of the fluorescence intensity of the mixture at a series of time points; wherein the phase shift between the modulated light source and the modulated fluorescent light is recorded and wherein decay times which differ from each other by less than a nanosecond up to 10 ns are deducible from the phase shift measurements ([0062], [0065]-[0073], [0084], [0086]), and quantifying the concentration of the protein of interest or of the conformational state of the protein of interest in the sample by reference to the fluorescence decay model (curve fit) ([0066]).
Lo et al. teaches photodetector temporal response (trigger system), as well as signal digitization and recording speed.
Lo et al. teaches excitation light in a wavelength of 300 nm and above, but does not teach using excitation light in a wavelength of 240-295 nm.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to conduct routine engineering optimization experimentation to determine excitation wavelengths to use for different proteins including wavelengths of 240 to 295 nm.
In paragraph [0042] Lo et al. also discusses how the time domain method provides significant information over shorter pulses.
As to the newly added limitation that the mixture is introduced into a flow cell wherein the mixture is exposed to one or more pulses of light, Kelleher et al. using fluorescent excitation pulses that are less than a nanosecond (480 picoseconds) to excite particles to be analyzed using decay times of the emission. (page 18, second paragraph and Figs. 15(a) and 15 (b)).
Kelleher et al. further teaches that detection can be accomplished using flow cytometry which renders it obvious to use a flow cell to differentiate proteins in a mixture that is passed through a flow cell and irritated with fluorescent radiation.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to differentiate proteins in mixtures via a flow cells, based on Kelleher et al. teaching flow cytometry for such analysis.
I.) As noted above, Lo et al. in view of Kelleher et al. renders obvious all of the element of claim 21.
Therefore, Lo et al. in view of Kelleher et al. renders claim 21 obvious.
II.) Regarding applicant’s claim 22, as noted above Lo et al. in view of Kelleher et al. renders claim 21 obvious from which claim 22 depends.
Claim 22 recites that the light source is a single light emitting diode, an array of light emitting diodes, and/or a laser; optionally wherein the laser is a diode pumped Q-switched solid state laser.
As noted above, Lo et al. teaches a laser light source.
Therefore, Lo et al. in view of Kelleher et al. renders claim 22 obvious.
III.) Regarding applicant’s claim 24, as noted above Lo et al. in view of Kelleher et al. renders claim 21 obvious from which claim 24 depends.
Claim 24 recites that the fluorescent emission is reflected towards the one or more detectors by an optionally ellipsoidal reflector or a lens; optionally wherein the fluorescent emission is reflected towards the one or more detectors via a filter assembly optionally comprising a long-pass optical filter.
Lo et al. teaches lens 160 that focuses fluorescent emission toward detector 130.
Therefore, Lo et al. in view of Kelleher et al. renders claim 24 obvious.
IV) Regarding applicant’s claim 30, as noted above Lo et al. in view of Kelleher et al. renders claim 21 obvious from which claim 30 depends.
Claim 30 recites that the one or more detectors are one or more photodiodes with a sub-nanosecond rise time; optionally wherein the one or more ultra-fast photodiodes is connected to a high bandwidth transimpedance amplifier.
Lo et al. teaches photodiode components. [0041]
Therefore, Lo et al. in view of Kelleher et al. renders claim 30 obvious.
V) Regarding applicant’s claim 36, as noted above Lo et al. in view of Kelleher et al. renders claim 21 obvious from which claim 36 depends.
Claim 36 recites that the light source is capable of addressing the mixture with pulses of light at a wavelength in the range 250-280 nm, optionally at a wavelength of 266 nm.
Lo et al, in view of Kelleher et al. does not teach that the light source is capable of addressing the mixture with pulses of light at a wavelength in the range 250-280 nm, optionally at a wavelength of 266 nm.
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to conduct routine engineering optimization experimentation to determine excitation wavelengths to use for different proteins including wavelengths in the range 250-280 nm, optionally at a wavelength of 266 nm.
Therefore, Lo et al. in view of Kelleher et al. renders claim 36 obvious.
6. Claim 28 is rejected under 35 U.S.C. 103 as being unpatentable over Lo et al. as applied to claim 21 above, and further in view of U.S. Patent Application Publication No. 2017/0290515 to Butte et al.
I.) Regarding applicant’s claim 28, as noted above Lo et al. renders claim 21 obvious from which claim 28 depends.
Claim 28 recites a beam splitter configured to split the emitted pulses of laser light into first and second portions, where the first portion is directed to a photodiode and the second portion is directed towards the protein of interest in the mixture of proteins, optionally wherein the trigger system is triggered by a signal from the photodiode.
Lo et al. does not teach a beam splitter configured to split the emitted pulses of laser light into first and second portions, where the first portion is directed to a photodiode and the second portion is directed towards the protein of interest in the mixture of proteins, optionally wherein the trigger system is triggered by a signal from the photodiode.
Butte et al. teaches a beam splitter that directs a portion of excitation light pulses to a photodiode that may be used to time delayed fluorescence signals. [0131]
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. to include a beam splitter configured to split the emitted pulses of laser light into first and second portions, where the first portion is directed to a photodiode as taught by Butte et al. and the second portion is directed towards the protein of interest in the mixture of proteins for purposes of using the photodiode to detect time-delayed fluorescence signals as taught by Butte et al.
7. Claim 32 is rejected under 35 U.S.C. 103 as being unpatentable over Lo et al. as applied to claim 21 above, and further in view of Applicant’s Admitted Prior Art.
I.) Regarding applicant’s claim 32, as noted above Lo et al. renders claim 21 obvious from which claim 32 depends.
Claim 32 recites a chromatography assembly, wherein said apparatus is assembled such that the mixture that is addressed by the light is eluate in an elution capillary of the chromatography assembly, optionally wherein the elution capillary is a straight capillary.
Lo et al. does not teach a chromatography column.
As noted above, in the Background section of the disclosure applicant notes that chromatography is commonly used is protein separation for bioprocessing. On page 2, lines 1-2 applicant nots that “In a typical industrial bioprocess, chromatography is the key step for purifying the product from other proteins, cellular debris and components of the media.”
It would have been obvious to one of ordinary skill in the art before applicant’s effective filing date to modify Lo et al. in view of Kelleher et al. to receive and quantify proteins from the eluate of a chromatography column as taught by Applicant’s Admitted Prior Art for purposes of bioprocessing and perform the quantifying of the proteins in the elute leaving the chromatography column. Providing an elution capillary from the chromatography would have been an obvious matter of size since such a size would be capable of the desired function.
Response to Arguments
Applicant's arguments filed 01/15/2026 have been fully considered but they are not persuasive.
On page 12 of applicant’s response applicant argues that Lo et al. teaches sinusoidal modulation of incident light.
As noted above, in paragraph [0042] Lo et al. teaches that pulsed light can be used. Therefore, Lo et al. teaches and otherwise renders pulsed light obvious.
Applicant’s argument that Lo et al. teaches analysis of solid tissue is not persuasive since Lo et al.’s teaching of differentiating proteins using pulsed fluorescence is applicable to detection/differentiating of other proteins.
Applicant’s argument’s that Lo et al. does not teach a flow cell is not persuasive since Kelleher et al. teaches detection can be accomplished using flow cytometry which renders it obvious to use a flow cell to differentiate proteins in a mixture that is passed through a flow cell and irradiated with fluorescent radiation.
On page 13 of applicant’s response applicant argues that Kelleher et al. teach distinguishing specific protein species.
As noted above, Lo et al. teaches differentiating proteins using pulsed fluorescence. Kelleher et al. has been relied upon as teaching the use of fluorescent excitation pulses that are less than a nanosecond (480 picoseconds) to excite particles to be analyzed using decay times of the emission.
The examiner notes that on page 14 of applicant’s response applicant notes that Lo et al. “does teach towards methods of measuring intrinsic fluorescent decay of proteins.” Although applicant’s point is that this teaching by Lo et al. is limited to proteins in solid tissues, one skilled in the art would recognize that this teaching would be obvious to apply to other types and sources of proteins.
On page 15 of applicant’s response applicant argues that Wheelock utilizes differences in fluorescent lifetimes to differentiate between fluorescence from specific protein labels and non-specific background fluorescence, and that Wheelock demonstrates time-resolved emission of the labels.
As noted above, Wheelock has been relied upon as teaching time-resolved emission of proteins that utilizes differences in fluorescent lifetime imaging to differentiate between fluorescence from specific protein labels and non-specific background fluorescence, and the use of deconvolution algorithms for multi-exponential decay, ([0044]) and using the entire area under the curve (AUC) for quantification purposes the AUC, corresponding to the fitted photon count for each specific component. [0051], rendering it obvious to modify Lo et al. in view of Kelleher et al. to use deconvolution of analysis the fluorescence decay curve to quantify the contribution of the intrinsic fluorescence decay signature from different proteins species or conformational states of the protein species to the fluorescence decay curve as taught by Wheelock.
On page 17 of applicant’s reply applicant argues that Schurzinger et al. does not teach that a background noise signal, I background (t), is calculated with reference to a baseline offset value and the width of the time window of a high-bandwidth digitizer or a sampling oscilloscope.
As noted above, Schurzinger et al. has been relied upon as teach digitizing signals and compensating for changes in background levels by monitoring amplitude variations, rendering it obvious to modify Lo et al. in view of Kelleher et al. to digitize signals and determine background levels by monitoring amplitude (width) variations.
The use of an appropriate baseline reference would have been obvious to one of ordinary skill in the art.
Conclusion
Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). 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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/M.S.G./Examiner, Art Unit 1798
/CHARLES CAPOZZI/Supervisory Patent Examiner, Art Unit 1798