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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on December 2nd, 2025 has been entered.
Response to Amendment
The Amendment filed December 2nd, 2025 has been entered. Examiner acknowledges the cancellation of claims 14, 17, 21, 25 and 29. Claims 12-13, 15-16, 18-20, 22-24, 26-28 and 30-36 remain pending in the application. Applicant’s amendments to the Claims have overcome the 112(b) rejections previously set forth in the Final Office Action mailed September 30th, 2025. Therefore, these rejections have been withdrawn. However, in light of these amendments, a new grounds of rejection is made in view of 35 USC § 112(b).
Response to Arguments
Applicant's arguments filed December 2nd, 2025 have been fully considered but they are not persuasive.
Regarding the rejection of claims 12-17, 21, 25 and 29 under 35 U.S.C. §103 over Ase (WO2019009295A1) in view of Cabral (WO2019068177A1) applicant has argued that the combination fails to render the amended claims obvious stating that Ase is directed to dialysis water systems and oligoDNA detection using a fluorometric method and does not teach or suggest fluorescent detection of endotoxins while asserting that Cabral, while disclosing fluorescent chemosensors for bacterial membrane components, is limited to laboratory-scale detection and does not disclose or suggest integration with industrial water purification systems. In response to applicant's arguments against the references individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). Further, Applicant’s arguments regarding pharmacopeial thresholds, specific detection limits, high-temperature operation, and other asserted “industrial” constraints are not commensurate with the scope of the pending claims, which do not recite a numerical detection limit, a required operating temperature, a minimum flowrate, or any other specific operational parameter establishing the asserted constraints.
Regarding the rejection of claims 18, 19, 22, 23, 26, 27, 30 and 31 under 35 U.S.C. § 103 over Ase in view of Cabral, and further in view of Kasai (""Staphylococcus aureus Detection by Fluorescent Silica Nanoparticles Modified with Metal Dipicolylamine Complexes") and Ozawa ("Effect of Cyclodextrins on Saccharide Sensing Function of a Fluorescent Phenylboronic Acid in Water") that Applicant has argued that the cited references fail to render the claims obvious as amended claims now expressly recite the use of dpa-HCC as the fluorescent substance and further specify its coordination with metal ions such as Cu2+, Ni2+, or Co2+, each of which is critical for enabling selective binding to phosphate groups present in endotoxins and neither Kasai nor Ozawa teaches or suggests the use of dpa-HCC specifically, nor its application in detecting endotoxins under stringent industrial water purification conditions. However, Kasai directly teaches fluorescent probes based on metal-dipicolylamine complexes including M-dpa-HCC, and describes their use for selective recognition of phosphate-containing biological targets in water. Thus, Kasai is highly relevant to the claimed limitations directed to a fluorescent substance having a dipicolylamino recognition site, coordination with metal ions (e.g., Cu2+/Ni2+/Co2+), and phosphate-related recognition behavior. Applicant’s assertions that dpa-HCC is “uniquely tailored” are conclusory argument absent objective evidence. The prior art already teaches the claimed probe motif (Kasai, see title and par. 1 of p. 749 “Cu-dpa-HCC/FSiNP”) and teaches endotoxin/LPS detection by fluorescence (Cabral par. [0008-0009]); combining known fluorescence recognition motifs to detect known biologically relevant negatively charged targets represents predictable use of prior art elements according to their established functions. Further, as above, applicant’s “industrial” performance arguments are not commensurate with the claims, which do not recite thresholds/operating constraints.
Regarding the rejection of claims 20, 24, 28 and 32 under 35 U.S.C. § 103 as being unpatentable over the same combination of Ase and Cabral, in further view of Ozawa applicant argues that the cited references do not render the amended claims obvious as the amended claims recite that the fluorescent substance includes a phenylboronic acid and new supporting claims 33-36 explicitly identify the compound as C1-APB. Further, Applicant argues that neither Cabral nor Ozawa teaches nor suggests using C 1-APB, an engineered fluorescent probe with an amide-bonded spacer, pyrene fluorophore, and phenylboronic acid recognition site, in the context of a pharmaceutical water production facility or method and Ozawa’s work focuses on saccharide sensing in solution. However, the rejection does not rely on Ozawa as expressly teaching endotoxin detection. Ozawa is relied upon for teaching phenylboronic-acid fluorescent probes such as C1-APB (an engineered fluorescent probe with an amide-bonded spacer, pyrene fluorophore, and phenylboronic acid recognition site, which Ozawa teaches in the abstract and throughout the publication may be used as a saccharide sensor) that bind saccharide/diol moieties in water and produce a measurable fluorescence response, and for explaining why boronic acid chemosensors are desirable (e.g., stability and strong fluorescence response). Cabral supplies the teaching of detecting endotoxin/LPS in aqueous samples and motivates selecting recognition chemistries that interact with endotoxin/LPS structural features. Because endotoxin/LPS includes saccharide/diol-containing moieties, it would have been obvious to apply Ozawa’s phenylboronic acid saccharide-recognition chemistry as an alternative recognition element within the combined Ase/Cabral framework. Again, as above, applicant’s “industrial” performance arguments are not commensurate with the claims, which do not recite thresholds/operating constraints.
Further applicant argues that the claimed invention requires highly specialized fluorescent probes uniquely suited for endotoxin detection in purified or injection water systems. However, the structure of the pending claims contradicts this assertion. The independent claims define two parallel claim families in which the same water-production and endotoxin-monitoring system is implemented using entirely different fluorescent recognition chemistries. For examples, claim 12 recites a fluorescent substance having a dipicolylamino recognition site coordinated with a metal ion, whereas claim 20 recites the same system employing a phenylboronic acid recognition site. Similarly, claims 13 and 24, claims 15 and 28, and claims 16 and 32 respectively recite otherwise identical system or method limitations differing only in the selected recognition chemistry. The claim structure therefore demonstrates that the claimed system is not limited to a particular specialized probe architecture, but instead encompasses multiple known fluorescent recognition motifs capable of interacting with structural features of endotoxin molecules. This supports the rationale that a person of ordinary skill in the art would have reasonably substituted known fluorescent recognition chemistries, such as those taught by the cited references, within the known water monitoring framework.
For the foregoing reasons, applicant’s arguments are not persuasive. The cited references collectively teach each claimed limitation, and the proposed combinations represent predictable substitutions of known detection chemistries/recognition motifs within known water-facility sampling and fluorescence monitoring networks, with a reasonable expectation of success.
Claim Objections
Claims 12, 24, 28 and 32 objected to because of the following informalities: drafting errors, typos. Appropriate correction is required.
Claim 12 recites “connecting the fluorescent site to the recognition site via spacer spacer…”. The phase “via spacer” lacks proper antecedent basis and should be amended to “via said spacer”.
Claims 24, 28 and 32 each recite phrases such as “said fluorescent substance is having a structure…”. The phrase “is having” is grammatically improper and should be amended for clarity (e.g., “has a structure” or “having a structure”).
Claims 28 and 32 each recite phrases such as: “subjecting a test subject sample to….detection of an endotoxin, said detection comprises detecting…”. This construction is awkward and should be revised to clarity (e.g., by combining the clauses or restructuring the sentence).
Claim 32 recites “wherein; said recognition site recognizes…”. The punctuation “wherein;” is improper and should be corrected.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 12, 13, 15 and 16 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claim 12, claim 12 recites in part: “ a supply unit configured to supply a fluorescent substance to the sample…” however, the claim previously introduces “a test subject sample”, and the phrase “the sample” lacks proper antecedent basis and renders the claim unclear. Additionally, claim 12 recites “said fluorescent substance has a dipicolylamino group represented by Formula 1, as spacer recognition site…”. The phrase “as spacer recognition site” is unclear because it is not evident whether the dipicolylamino group itself constitutes the recognition site or whether it is associated with a spacer that forms part of the recognition site. Therefore, the metes and bounds of the claim are unclear.
Regarding claim 13, reciting that the sample collection unit collects the test subject sample “directly from spacer injection water tank…” lacks a clear meaning in the context of the claim. It is unclear whether this phrase was intended to refer to the previously recited injection water tank or some other structure.
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.
Claims 12-13, 15-16, 18-19, 22-23, 26-27, and 30-31 are rejected under 35 U.S.C. 103 as being unpatentable over Ase (WO-2019009295-A1, an English machine translation is provided with this office action and is used for claim mapping in the prior art rejection below) in view of Cabral (WO-2019068177-A1), and further in view of Kasai ""Staphylococcus aureus Detection by Fluorescent Silica Nanoparticles Modified with Metal Dipicolylamine Complexes".
Regarding claim 12, Ase discloses a purified water production facility, comprising: a purified water production unit that obtains purified water from raw water (Ase Fig. 1 "raw water line" #s 11-13) via at least one of a reverse osmosis membrane device (Ase abstract and Fig. 1, #5) that performs reverse osmosis filtration of the raw water or an ion exchange device that performs ion exchange of the raw water; a sample collection unit that collects the test subject sample from the purified water (Ase Fig. 1 #19 "No. 2 Intake line"); and an endotoxin detection device (Ase par. 6 of p. 5 "ET concentration measuring device"). Ase also discloses collecting samples and the analysis of collected samples by performing fluorescence measurement of the water sample.
Ase however, does not explicitly disclose the endotoxin detection device comprising: a sample introduction unit into which a test subject sample is introduced; a supply unit configured to supply a fluorescent substance to the sample, said fluorescent substance having a structure in which a fluorescent site and a recognition site that recognizes a specific site of a molecular structure of an endotoxin are connected by a spacer; a reaction unit in which the sample and said fluorescent substance react with each other; and a detection unit configured to detect light emission or color development of the fluorescent substance subjected to the reaction, wherein: said spacer in said fluorescent substance has from 1 to 10 carbon atoms. and each chemical bond of a straight chain connecting the fluorescent site to the recognition site via said spacer is only a single bond; and said fluorescent substance has a dipicolylamino group represented by Formula 1, as spacer recognition site to which a metal ion Mn+, wherein n is a natural number, is coordinated:
[Formula 1]
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Cabral teaches performing fluorescence-based detection of bacterial membrane components including lipopolysaccharide (LPS) using a fluorescent probe assay in which a sample is introduced into a well plate (Cabral "96-well plate" par. [0200]), mixed with the fluorescent probe, allowed to react (Cabral par. [0200]), and analyzed by a fluorescence detector (Cabral par. [0124] "Tecan Infinite M1000"). Cabral teaches the fluorescent substance having a structure (Cabral par. [0200] claims 1 and 24) in which a fluorescent site and a recognition site that recognizes a specific site of a molecular structure of an endotoxin are connected by a spacer (Cabral claim 1 "linker"); and a detection unit (Cabral par. [0124] "Tecan Infinite M1000") configured to detect light emission or color development of the fluorescent substance subjected to the reaction.
Cabral further teaches fluorescent probes comprising dipicolylamine metal complexes such as Zn2+-DPA structures which function as recognition sites interacting with biological molecular structures.
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(Cabral p. 80)
Cabral also teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains (Cabral pars. [0086-0088]). Such linkers described by Cabral include spacers having between one and ten carbon atoms and consisting of single-bond carbon chains, corresponding to the claimed spacer connecting the fluorescent site and recognition site.
Kasai teaches fluorescent probes comprising a fluorescent moiety and a dipicolylamine (DPA) recognition site coordinated with a metal ion, including probes in which the fluorescent site and recognition site are connected by a spacer structure. Kasai further teaches that such metal-dipicolylamine complexes interact with biologically relevant phosphate-containing structures, producing fluorescence responses suitable for detection in aqueous environments.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to use the fluorescent endotoxin detection approach taught by Cabral within the purified-water monitoring system of Ase in order to monitor endotoxin contamination in purified water samples produced by the purification facility. A person of ordinary skill in the art would have recognized that fluorescent probe assays provide rapid and sensitive detection of endotoxin in aqueous samples and therefore would have been suitable for monitoring purified water systems. It would have further been obvious to employ the metal-dipicolylamine recognition motif taught by Kasai as the fluorescent probe because Kasai teaches that such probes bind biologically relevant molecular structures in aqueous environments and generate detectable fluorescence responses.
Regarding claim 13, Ase, in view of Cabral and further in view of Kasai discloses an injection water production facility, comprising: a purified water production unit that obtains purified water from raw water (Ase Fig. 1 "raw water line" #s 11-13) via at least one of a reverse osmosis membrane device (Ase abstract and Fig. 1, #5) that performs reverse osmosis filtration of the raw water or an ion exchange device that performs ion exchange of the raw water; an injection water production unit (Ase Fig. 1 #7 "Ultrafiltration membrane device") that obtains injection water from the purified water via a polymer membrane filtration device (Ase p.6 line 10 "FS 10 FC-FUST 653" has a polyethersulfone (PES) membrane) that filters the purified water or a distiller that distills the purified water; an injection water tank that maintains and stores said injection water in a heated state (Ase p.6 line 23 and #6a of Fig. 1); a delivery line that delivers said injection water provided in said injection water tank to a predetermined place of use (Ase teaches collecting samples from the water distribution system using intake lines (Ase Fig. 1 #18 "intake line"), which are connected to the purified/injection water distribution lines downstream of the treatment units. A person of ordinary skill in the art would have understood that such sampling points may be located along delivery lines or near storage tanks within the distribution system); a sample collection unit that collects a test subject sample from the injection water (Ase Fig. 1 #19 "No. 2 Intake line"); and an endotoxin detection device (Ase par. 6 of p. 5) "ET concentration measuring device") comprising: a sample introduction unit into which said test subject sample is introduced (Cabral par. [0200] teaches fluorescence-based detection of bacterial membrane components including lipopolysaccharide (LPS) using a fluorescent probe assay in which a sample is introduced into a “96- well plate”); a supply unit configured to supply a fluorescent substance to said test subject sample (Cabral teaches combining a volume of the sensor (supplying the fluorescence substance) with the analyte in a “384-well plate”), said fluorescent substance having a structure (Cabral par. [0200] claims 1 and 24) in which a fluorescent site and a recognition site that recognizes a specific site of a molecular structure of an endotoxin are connected by a spacer (Cabral claim 1 "linker"); a reaction unit in which said test subject sample and said fluorescent substance react with each other (Cabral par. [0200] teaches incubating the mixture for 10 minutes away from light indicating a reaction unit in which the fluorescent probe interacts with the analyte); and a detection unit configured to detect light emission or color development of the fluorescent substance subjected to the reaction (Cabral par. [0124] "Tecan Infinite M1000"), wherein: said sample collection unit collects said test subject sample directly from said injection water tank or from the delivery line at a downstream side of the injection water production unit and at an upstream side of the injection water tank (Ase teaches collecting samples from the injection water distribution system using intake lines connected to the water supply lines (Ase Fig. 1 #19 “No. 2 intake line”) These intake lines obtain samples from the injection water flowing through the distribution network including storage tanks and delivery lines (Ase Fig. 1 #6a injection water tank and #18 delivery line)); said spacer in said fluorescent substance has from 1 to 10 carbon atoms, and each chemical bond of a straight chain connecting the fluorescent site to the recognition site via said spacer is only a single bond (Cabral par. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains within 1 to 10 carbon atoms); and said fluorescent substance has a dipicolylamino group represented by Formula 2 (Cabral p. 80 teaches fluorescent probe incorporating dipicolylamino groups. Kasai par. 1 and Fig. 1 likewise teaches fluorescent probes comprising a metal-coordinated dpa “M-dpa-HCC”), as the recognition site to which a metal ion Mn+, wherein n is a natural number, is coordinated:
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Regarding claim 15, Ase, in view of Cabral and further in view of Kasai discloses a method of producing purified water, the method comprising: treating raw water (Ase Fig. 1 "raw water line" #s 11-13) by at least one of reverse osmosis filtration or ion exchange to obtain purified water (Ase abstract and Fig. 1, #5); and subjecting a test subject sample collected from said purified water to detection of an endotoxin (Ase par. 6 of p. 5 "ET concentration measuring device"), using a fluorescent substance that comprises a fluorescent site and a recognition site (Cabral par. [0200] and claims 1 and 24) connected by a spacer (Cabral claim 1 "linker"), wherein: said recognition site recognizes a specific site of a molecular structure of said endotoxin (Cabral claim 1(b) and par. [0009] “lipopolysaccharides (LPS)… of gram negative bacteria”), said spacer in said fluorescent substance has from 1 to 10 carbon atoms (Cabral pars. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains), and each chemical bond of a straight chain connecting the fluorescent site to said recognition site via said spacer is only a single bond (Cabral pars. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains within 1 to 10 carbon atoms); and said fluorescent substance has a dipicolylamino group represented by Formula 3 (Cabral p. 80 teaches use of a dipicolylamino group as does Kasai par. 1 and Fig. 1 “M-dpa-HCC”), as said recognition site to which a metal ion Mn+, wherein n is a natural number, is coordinated:
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Regarding claim 16, Ase, in view of Cabral and further in view of Kasai discloses a method of producing injection water, the method comprising: treating raw water (Ase Fig. 1 "raw water line" #s 11-13) by at least one of reverse osmosis filtration or ion exchange to obtain purified water (Ase abstract and Fig. 1, #5); obtaining injection water from said purified water by filtration via polymer membrane filtration or by distillation (Ase p.6 line 10 "FS 10 FC-FUST 653" has a polyethersulfone (PES) membrane); and subjecting a test subject sample collected from the injection water to detection of an endotoxin (Ase par. 6 of p. 5 "ET concentration measuring device"), using a fluorescent substance that comprises a fluorescent site and a recognition site (Cabral par. [0200] and claims 1 and 24) connected by a spacer (Cabral claim 1 "linker"), wherein: the recognition site recognizes a specific site of a molecular structure of said endotoxin (Cabral claim 1(b) and par. [0009] “lipopolysaccharides (LPS)… of gram negative bacteria”), said spacer in said fluorescent substance has from 1 to 10 carbon atoms, and each chemical bond of a straight chain connecting said fluorescent site to said recognition site via said spacer is only a single bond (Cabral pars. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains within 1 to 10 carbon atoms); and said fluorescent substance has a dipicolylamino group represented by Formula 4 (Cabral p. 80 teaches use of a dipicolylamino group as does Kasai par. 1 and Fig. 1 “M-dpa-HCC”), as said recognition site to which a metal ion Mn+. wherein n is a natural number. is coordinated:
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Regarding claim 18, Ase, in view of Cabral and further in view of Kasai discloses the purified water production facility according to claim 12, wherein said fluorescent substance is dpa-HCC represented by Formula 5 (Kasai par. 1 and Fig. 1 “M-dpa-HCC”), having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site, and wherein said spacer has one carbon atom, and said specific site is a phosphate group of said endotoxin (Cabral par. [0009] teaches fluorescent sensing systems for detecting gram-negative bacteria and further explains that lipopolysaccharide is a major component of gram-negative bacterial membranes. Cabral par.[0091] additionally teaches that binding occurs between positively charged metal ions and negatively charged phosphate-containing membrane components. These teachings indicate that the sensing system interacts with phosphate-containing sites associated with gram-negative bacterial membrane components, including LPS which is a key component of endotoxin):
[Formula 5]
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Regarding claim 19, Ase, in view of Cabral and further in view of Kasai discloses the purified water production facility according to claim 18, wherein the metal ion is a copper ion (Cu2+), a nickel ion (Ni2+), or a cobalt ion (Co2+) (Kasai last par. of left col. on p. 750 teaches that the metal-dpa probes may include transition metals such as Ni2+, Cu2+ and Zn2+).
Regarding claim 22, Ase, in view of Cabral and further in view of Kasai discloses the injection water production facility according to claim 13, wherein the fluorescent substance is dpa-HCC represented by Formula 7 (Kasai Fig. 1A), having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site, and wherein said spacer has one carbon atom, and said specific site is a phosphate group of an endotoxin (Cabral par. [0009] teaches fluorescent sensing systems for detecting gram-negative bacteria and further explains that lipopolysaccharide is a major component of gram-negative bacterial membranes. Cabral par.[0091] additionally teaches that binding occurs between positively charged metal ions and negatively charged phosphate-containing membrane components. These teachings indicate that the sensing system interacts with phosphate-containing sites associated with gram-negative bacterial membrane components, including LPS which is a key component of endotoxin):
[Formula 7]
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Regarding claim 23, Ase, in view of Cabral and further in view of Kasai discloses the injection water production facility according to claim 22, wherein the metal ion is a copper ion (Cu2+), a nickel ion (Ni2+), or a cobalt ion (Co2+) (Kasai last par. of left col. on p. 750 teaches that the metal-dpa probes may include transition metals such as Ni2+, Cu2+ and Zn2+).
Regarding claim 26, Ase, in view of Cabral and further in view of Kasai discloses the method of producing purified water according to claim 15 (Kasai Fig. 1A), wherein the fluorescent substance is dpa-HCC represented by Formula 9 having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site, and wherein said spacer has one carbon atom, and the specific site is a phosphate group of an endotoxin (Cabral par. [0009] teaches fluorescent sensing systems for detecting gram-negative bacteria and further explains that lipopolysaccharide is a major component of gram-negative bacterial membranes. Cabral par.[0091] additionally teaches that binding occurs between positively charged metal ions and negatively charged phosphate-containing membrane components. These teachings indicate that the sensing system interacts with phosphate-containing sites associated with gram-negative bacterial membrane components, including LPS which is a key component of endotoxin):
[Formula 9]
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Regarding claim 27, Ase, in view of Cabral and further in view of Kasai discloses the method of producing purified water according to claim 26, wherein the metal ion is a copper ion (Cu2+), a nickel ion (Ni2+), or a cobalt ion (Co2+) (Kasai last par. of left col. on p. 750 teaches that the metal-dpa probes may include transition metals such as Ni2+, Cu2+ and Zn2+).
Regarding claim 30, Ase, in view of Cabral and further in view of Kasai discloses the method of producing injection water according to claim 16, wherein the fluorescent substance is dpa-HCC represented by Formula 11 having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site, and wherein the spacer has one carbon atom (Kasai Fig. 1A), and the specific site is a phosphate group of an endotoxin (Cabral par. [0009] teaches fluorescent sensing systems for detecting gram-negative bacteria and further explains that lipopolysaccharide is a major component of gram-negative bacterial membranes. Cabral par.[0091] additionally teaches that binding occurs between positively charged metal ions and negatively charged phosphate-containing membrane components. These teachings indicate that the sensing system interacts with phosphate-containing sites associated with gram-negative bacterial membrane components, including LPS which is a key component of endotoxin):
[Formula 11]
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Regarding claim 31, Ase, in view of Cabral and further in view of Kasai discloses the method of producing injection water according to claim 30, wherein the metal ion is a copper ion (Cu2+), a nickel ion (Ni2+), or a cobalt ion (Co2+) (Kasai last par. of left col. on p. 750 teaches that the metal-dpa probes may include transition metals such as Ni2+, Cu2+ and Zn2+).
Claims 20, 24, 28, and 32-36 are rejected under 35 U.S.C. 103 as being unpatentable over Ase (WO-2019009295-A1, an English machine translation is provided with this office action and is used for claim mapping in the prior art rejection below) in view of Cabral (WO-2019068177-A1), and further in view of Ozawa, "Effect of Cyclodextrins on Saccharide Sensing Function of a Fluorescent Phenylboronic Acid in Water".
Regarding claim 20, Ase discloses a purified water production facility comprising: a purified water production unit that obtains purified water from raw water (Ase Fig. 1 "raw water line" #s 11-13) via at least one of a reverse osmosis membrane device (Ase abstract and Fig. 1, #5) that performs reverse osmosis filtration of the raw water or an ion exchange device that performs ion exchange of the raw water; a sample collection unit that collects the test subject sample from the purified water (Ase Fig. 1 #19 "No. 2 Intake line"); and an endotoxin detection device (Ase par. 6 of p. 5 "ET concentration measuring device"). Ase also discloses collecting samples and the analysis of collected samples by performing fluorescence measurement of the water sample.
Ase, however, does not expressly disclose the endotoxin device comprising: a sample introduction unit into which a test subject sample is introduced; a supply unit configured to supply a fluorescent substance to the sample, the fluorescent substance having a structure in which a fluorescent site and a recognition site that recognizes a specific site of a molecular structure of an endotoxin are connected by a spacer; a reaction unit in which the sample and the fluorescent substance react with each other; and a detection unit configured to detect light emission or color development of the fluorescent substance subjected to the reaction, wherein: the spacer in the fluorescent substance has from 1 to 10 carbon atoms, and each chemical bond of a straight chain connecting said fluorescent site to said recognition site via said spacer is only a single bond; and said fluorescent substance has phenylboronic acid represented by Formula 6, as said recognition site,
[Formula 6]
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Cabral teaches performing fluorescence-based detection of bacterial membrane components including lipopolysaccharide (LPS) using a fluorescent probe assay in which a sample is introduced into a well plate (Cabral "96-well plate" par. [0200]), mixed with the fluorescent probe, allowed to react (Cabral par. [0200]), and analyzed by a fluorescence detector (Cabral par. [0124] "Tecan Infinite M1000"). Cabral teaches the fluorescent substance having a structure (Cabral par. [0200] claims 1 and 24) in which a fluorescent site and a recognition site that recognizes a specific site of a molecular structure of an endotoxin are connected by a spacer (Cabral claim 1 "linker"); and a detection unit (Cabral par. [0124] "Tecan Infinite M1000") configured to detect light emission or color development of the fluorescent substance subjected to the reaction.
Cabral also teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains (Cabral pars. [0086-0088 Such linkers described by Cabral include spacers having between one and ten carbon atoms and consisting of single-bond carbon chains, corresponding to the claimed spacer connecting the fluorescent site and recognition site.
Ozawa teaches fluorescent phenylboronic-acid chemosensors (e.g., C1-APB) for saccharide/diol recognition in water, including a probe structure having (i) a phenylboronic acid recognition site, (ii) a pyrene fluorescent site, and (iii) a spacer including an amide linkage that improves aqueous compatibility. The phenylboronic acid moiety of Ozawa corresponds to the claimed recognition site represented by formula 6, which binds saccharide/diol structures in aqueous environments. Ozawa further explains that boronic-acid based chemosensors are attractive due to their stability and that the disclosed C1-APB system provides high affinity to saccharides and a substantial fluorescence response upon saccharide binding.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to apply Ozawa’s phenylboronic-acid saccharide-recognition chemistry as an alternative recognition element within the combined Ase/Cabral fluorescence detection framework. Ase supplies the purified water monitoring environment and fluorescence-based monitoring context, while Cabral teaches fluorescence-based detection of bacterial membrane components including lipopolysaccharide (LPS). Because LPS contains carbohydrate structures and phenylboronic acids are well known to bind saccharide and diol groups, a person of ordinary skill in the art would have recognized that Ozawa’s phenylboronic-acid probes would interact with saccharide portions of LPS and produce a detectable fluorescence response. Substituting this known saccharide-recognition chemistry into the known fluorescence detection platform would have represented the predictable use of prior-art elements according to their established functions, with a reasonable expectation of success.
Regarding claim 24, Ase, in view of Cabral and further in view of Ozawa discloses an injection water production facility comprising: a purified water production unit that obtains purified water from raw water (Ase Fig. 1 "raw water line" #s 11-13) via at least one of a reverse osmosis membrane device (Ase abstract and Fig. 1, #5) that performs reverse osmosis filtration of said raw water or an ion exchange device that performs ion exchange of said raw water; an injection water production unit (Ase Fig. 1 #7 "Ultrafiltration membrane device") that obtains injection water from said purified water via a polymer membrane filtration device(Ase p.6 line 10 "FS 10 FC-FUST 653" has a polyethersulfone (PES) membrane) that filters said purified water or a distiller that distills said purified water; an injection water tank that maintains and stores said injection water in a heated state (Ase p.6 line 23 and #6a of Fig. 1); a delivery line that delivers said injection water provided in said injection water tank to a predetermined place of use (Ase Fig. 1 #18 "intake line"); a sample collection unit that collects a test subject sample from said injection water (Ase Fig. 1 #19 "No. 2 Intake line"); and an endotoxin detection device (Ase par. 6 of p. 5) "ET concentration measuring device") that comprises: a sample introduction unit into which said test subject sample is introduced (Cabral par. [0200] teaches performing endotoxin detection using a fluorescent probe assay in which a sample is introduced into a “96- well plate”); a supply unit configured to supply a fluorescent substance to the sample (Cabral teaches combining a volume of the sensor (supplying the fluorescence substance) with the analyte in a “384-well plate”), said fluorescent substance is having a structure (Cabral par. [0200] claims 1 and 24) in which a fluorescent site and a recognition site that recognizes a specific site of a molecular structure of an endotoxin are connected by a spacer (Cabral claim 1 "linker"); a reaction unit in which said test subject sample and said fluorescent substance react with each other (Cabral par. [0200] teaches that "mixtures were incubated for 10 min away from light" indicating a unit where a reaction took place); and a detection unit configured to detect light emission or color development of said fluorescent substance subjected to the reaction (Cabral par. [0124] "Tecan Infinite M1000"), wherein: said sample collection unit collects said test subject sample directly from said injection water tank or from said delivery line at a downstream side of said injection water production unit and at an upstream side of said injection water tank (Ase teaches collecting samples from the injection water distribution system using intake lines connected to the water supply lines (Ase Fig. 1 #19 “No. 2 intake line”) These intake lines obtain samples from the injection water flowing through the distribution network including storage tanks and delivery lines (Ase Fig. 1 #6a injection water tank and #18 delivery line)); said spacer in said fluorescent substance has from 1 to 10 carbon atoms, and each chemical bond of a straight chain connecting said fluorescent site to said recognition site via said spacer is only a single bond (Cabral par. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains within 1 to 10 carbon atoms); and, said fluorescent substance has phenylboronic acid represented by Formula 8 as said recognition site,
[Formula 8]
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(Ozawa title “fluorescent phenylboronic acid”).
Regarding claim 28, Ase, in view of Cabral and further in view of Ozawa discloses a method of producing purified water comprising: treating raw water (Ase Fig. 1 "raw water line" #s 11-13) by at least one of reverse osmosis filtration or ion exchange to obtain purified water(Ase abstract and Fig. 1, #5); and subjecting a test subject sample collected from said purified water to detection of an endotoxin (Ase par. 6 of p. 5 "ET concentration measuring device"), said detection comprises detecting an endotoxin from said test subject sample using a fluorescent substance having a structure in which a fluorescent site and a recognition site (Cabral par. [0200] and claims 1 and 24) are connected by a spacer (Cabral claim 1 "linker"), wherein: said recognition site recognizes a specific site of a molecular structure of said endotoxin (Cabral claim 1(b) and par. [0009] “lipopolysaccharides (LPS)… of gram negative bacteria”); said spacer in said fluorescent substance has from 1 to 10 carbon atoms (Cabral pars. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains within 1 to 10 carbon atoms), and each chemical bond of a straight chain connecting said fluorescent site to said recognition site via said spacer is only a single bond (Cabral pars. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers alkyl (single-bond) chains); and said fluorescent substance is having phenylboronic acid represented by Formula 10 as said recognition site,
[Formula 10]
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(Ozawa “C1-APB” p. 207 and Fig. 1).
Regarding claim 32, Ase, in view of Cabral and further in view of Ozawa discloses a method of producing injection water comprising: treating raw water (Ase Fig. 1 "raw water line" #s 11-13) by at least one of reverse osmosis filtration or ion exchange to obtain purified water (Ase abstract and Fig. 1, #5); obtaining injection water from said purified water by filtration via polymer membrane filtration of said purified water or by distillation of said purified water (Ase p.6 line 10 "FS 10 FC-FUST 653" has a polyethersulfone (PES) membrane); and subjecting a test subject sample collected from said injection water to detection of an endotoxin (Ase par. 6 of p. 5 "ET concentration measuring device"), said detection comprises detecting said endotoxin from said test subject sample using a fluorescent substance having a structure in which a fluorescent site and a recognition site (Cabral par. [0200] and claims 1 and 24) are connected by a spacer (Cabral claim 1 "linker"), wherein; said recognition site recognizes a specific site of a molecular structure of said endotoxin (Cabral claim 1(b) and par. [0009] “lipopolysaccharides (LPS)… of gram negative bacteria”); said spacer in said fluorescent substance has from 1 to 10 carbon atoms, and each chemical bond of a straight chain connecting said fluorescent site to said recognition site via said spacer is only a single bond (Cabral pars. [0086-0088] teaches fluorescent probes in which the fluorescent moiety and recognition group are connected by linkers including short alkyl chains within 1 to 10 carbon atoms) ; and said fluorescent substance is having phenylboronic acid represented by Formula 12 as said recognition site,
[Formula 12]
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(Ozawa title “fluorescent phenylboronic acid”).
Regarding claim 33, Ase, in view of Cabral and further in view of Ozawa discloses the purified water production facility according to claim 20, wherein the fluorescent substance is C1-APB represented by Formula 13 having pyrene as said fluorescent site, and wherein said spacer has an amide bond, and said specific site is a sugar chain moiety of said endotoxin:
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(Ozawa “C1-APB” p. 207 and Fig. 1).
Regarding claim 34, Ase, in view of Cabral and further in view of Ozawa discloses the injection water production facility according to claim 24, wherein said fluorescent substance is C1-APB represented by Formula 14 having pyrene as said fluorescent site, and wherein said spacer has an amide bond, and said specific site is a sugar chain moiety of said endotoxin:
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(Ozawa “C1-APB” p. 207 and Fig. 1).
Regarding claim 35, Ase, in view of Cabral and further in view of Ozawa discloses the method of producing purified water according to claim 28, wherein the fluorescent substance is C1-APB represented by Formula 15 having pyrene as said fluorescent site, and wherein said spacer has an amide bond, and said specific site is a sugar chain moiety of said endotoxin:
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(Ozawa “C1-APB” p. 207 and Fig. 1).
Regarding claim 36, Ase, in view of Cabral and further in view of Ozawa discloses the method of producing injection water according to claim 32, wherein the fluorescent substance is C1-APB represented by Formula 16 having pyrene as said fluorescent site, and wherein said spacer has an amide bond, and said specific site is a sugar chain moiety of said endotoxin:
[Formula 16]
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(Ozawa “C1-APB” p. 207 and Fig. 1).
Conclusion
Any inquiry concerning this communication or earlier communications from the examiner should be directed to WILLIAM ADDISON GEISBERT whose telephone number is (703)756-5497. The examiner can normally be reached Mon-Fri 7:30-5:00 EDT.
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/W.A.G./ Examiner, Art Unit 1779
/Bobby Ramdhanie/ Supervisory Patent Examiner, Art Unit 1779