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 .
Response to Amendment
The Amendment filed June 17, 2026 has been entered. Claims 12-13, 15-16, 18-20, 22-24, 26-28, and 30-52 remain pending in the application. Applicant’s amendments to the Claims have overcome each of the objections and rejections under 35 U.S.C. § 112(b) previously set forth in the previous Office Action mailed March 19, 2026. Therefore, the objections and rejections have been withdrawn. The amendments materially alter the limitations upon which the previous rejections under 35 U.S.C. § 103 were based. Accordingly, the previous 103 rejections are withdrawn and new grounds of rejection are set forth below in view of the claims as presently amended and the additional prior art made of record.
Response to Arguments
Applicant's arguments filed June 17, 2026, including the subsequently submitted supporting literature, have been fully considered. The arguments are persuasive to the extent that the prior rejection did not expressly address the newly added limitations requiring quantification of endotoxin from 0.01 EU/mL to 10 EU/mL and, with respect to claims 13 and 16, repeated measurements at intervals of 30 minutes or less. However, the arguments are not persuasive as to the patentability of the claims as presently amended for reasons discussed below.
Applicant argues that Cabral does not disclose quantitative endotoxin detection within the newly claimed 0.01-10 EU/mL range and that its disclosed experimental concentrations are substantially higher than the claimed range. The Examiner acknowledges that Cabral alone does not supply this newly added limitation. The rejection below, however, additionally relies upon Bonen (US-20050124013-A1). Bonen expressly teaches automated on-line endotoxin analysis of fluids from production lines, including water and pharmaceutical products, and teaches determining endotoxin concentrations with good accuracy and reproducibility over the range of 0.01 to 10 EU/mL. Bonen further expressly contemplates fluorescence measurement and fluorogenic detection. Thus, Applicant’s argument concerning the concentration range disclosed by Cabral does not distinguish the combination of references presently relied upon.
Applicant further argues that obtaining quantitative fluorescence measurements at the claimed low endotoxin concentration would not have constituted routine optimization and relies upon the submitted literature concerning fluorescence detection limits. This evidence has been considered. However, it does not overcome the express pre-filing teaching of Bonen that quantitative endotoxin determination over the claimed 0.01-10 E/mL range was known in automated endotoxin analysis, together with Bonen’s express disclosure of fluorogenic and fluorescence-based embodiments.
Regarding claims 13 and 16, Applicant argues that Cabral’s incubation and subsequent fluorescence measurement do not teach repeatedly measuring the test subject sample at intervals of 30 minutes or less. The Examiner agrees that Cabral alone does not expressly disclose this newly added limitation. Lorch (US-20010040130-A1), however, expressly teaches continuous and real-time monitoring of endotoxin in high-purity water, including water for injection, and describes on-line monitoring integrated into a high purity water system. Such continuous monitoring necessarily provides repeated measurements at intervals shorter than 30 minutes. Accordingly, this argument does not distinguish the combination of references presently relied upon.
For these reasons, Applicant’s arguments do not overcome the rejections set forth below.
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, 15, 49 and 51 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 Bonen (US-20050124013-A1).
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 a test subject sample from the purified water (Ase Fig. 1 #19 "No. 2 Intake line"); and an endotoxin detection device comprising (Ase par. 6 of p. 5 "ET concentration measuring device"). Ase further discloses collecting samples and analyzing the collected samples by fluorescence measurement of the water sample.
Ase, however does not explicitly disclose the endotoxin device comprising: a sample introduction unit into which said test subject sample is introduced; a supply unit configured to supply a fluorescent substance to said test subject 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 said test subject sample and said fluorescent substance react with each other; and a detection unit configured to detect light emission or color development of said 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 said fluorescent site to said recognition site via said spacer is only a single bond; and said endotoxin detection device is configured to quantify an endotoxin concentration in a range of from 0.01 EU/mL to 10 EU/mL.
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 M 1000"). Cabral further discloses fluorescent probes having a fluorescent moiety and a recognition moiety connected by a linker (Cabral claim 1 "linker"), corresponding to a fluorescent site and a recognition site connected by a spacer (Cabral par. [0200] claims 1 and 24), and discloses linkers including short alkyl chains having from 1 to 10 carbon atoms and consisting of single-bond carbon chains, corresponding to the claimed spacer connecting the fluorescent site and the recognition site (Cabral pars. [0086-0088]).
Bonen discloses automated endotoxin detection systems for testing fluid samples from production lines during the preparation of, among other things, water, pharmaceutical products, and parenteral preparations, wherein a test fluid sample is mixed with endotoxin-detection agents and the level of endotoxin concentration is determined. Bonen expressly discloses that the automated system determines endotoxin concentration with good accuracy and reproducibility over a range of 0.01 to 10 EU/mL (Bonen par. [0047]). Bonen further discloses fluorescence testing (Bonen par. [0089]) and fluorogenic detection and specifically contemplates endotoxin monitoring of water obtained from a Water For Injection (WFI) or high-purity-water process loop.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to employ the fluorescence-based endotoxin/LPS detection approach taught by Cabral within the purified-water monitoring system of Ase and to configure the resulting endotoxin detection device to quantitatively determine endotoxin concentration over the range of 0.01 EU/mL to 10 EU/mL, as taught by Bonen. Ase already provides a purified water production and sampling environment in which endotoxin concentration is monitored, Cabral provides a known fluorescence-based sensing arrangement for detecting LPS using a sample, fluorescent sensing substance, reaction, and fluorescence detector, and Bonen expressly demonstrates quantitative endotoxin monitoring over the claimed concentration range in production-line fluids including WFI and high-purity water. A person of ordinary skill in the art therefore would have had reason to incorporate Cabral’s fluorescence-based sensing arrangement and Bonen’s known quantitative operating range into Ase’s purified-water monitoring system in order to provide rapid, sensitive, accurate, and reproducible quantitative monitoring of endotoxin contamination in purified water, with a reasonable expectation of success.
Regarding claim 15, Ase in view of Cabral and further in view of Bonen 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 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 detection of said endotoxin comprises quantifying an endotoxin concentration of from 0.01 EU/mL to 10 EU/mL (Bonen par. [0047]).
Regarding claim 49, Ase in view of Cabral and further in view of Bonen discloses the purified water production facility according to claim 12, wherein said endotoxin detection device is configured to perform online analysis, offline analysis, or both (Bonen pars. [0046] and [0055]).
Regarding claim 51, Ase in view of Cabral and further in view of Bonen discloses the method of producing purified water according to claim 15, wherein said detection of said endotoxin is performed by online analysis, offline analysis, or both (Bonen pars. [0046] and [0055]).
Claims 13, 16, 50 and 52 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 Bonen (US-20050124013-A1), and further in view of Lorch (US-20010040130-A1).
Regarding claim 13, Ase 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 #6 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 with in the distribution system); a sample collection unit that collects a test subject sample from the injection water (Ase Fig. 1 #19 "Intake line"); and an endotoxin detection device (Ase par. 6 of p. 5) "ET concentration measuring device"). Ase further teaches heating the water within its circulation system to about 80-85°C during the disclosed sterilization operation and sampling the treated water produced by the system. (Ase Example 1 and water tank 6, heater 6a, UF device 7, intake lines 18-20 and the associated water lines.)
Ase, however, does not explicitly disclose that the endotoxin device comprises a sample introduction unit into which the test subject sample is introduced; a supply unit configured to supply a fluorescent substance to said test subject 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 said test subject sample and said fluorescent substance react with each other; and a detection unit configured to detect light emission or color development of said fluorescent substance subjected to the reaction, 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; 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; said endotoxin detection device is configured to quantify an endotoxin concentration in a range of from 0.01 EU/mL to 10 EU/mL; and said endotoxin detection device is configured to repeatedly measure said test subject sample at measurement intervals of 30 minutes or less.
Cabral teaches fluorescence-based detection of bacterial membrane components including lipopolysaccharide (LPS) using fluorescent sensor compounds having a fluorophore connected through a link to a recognition/metal-ion-coordinating moiety. Cabral teaches introducing an analyte sample and fluorescent sensor into a well, allowing the sensor and analyte to interact, and measuring the resulting fluorescence using a fluorescence detector (Cabral par. [0124], Tecan Infinite M1000; see also the LPS/bacterial fluorescence experiments). Cabral further expressly identifies lipopolysaccharide among the negatively charged phosphate-containing membrane components that may be detected by its fluorescence methods. Cabral also teaches link moieties that may comprise saturated alkylene chains, thereby teaching fluorescent compounds in which the fluorophore and recognition moiety are connected through short carbon-containing linkers corresponding to the recited space architecture.
Bonen further teaches automated endotoxin detection in water-production systems and expressly teaches the newly recited quantitative range. Specifically, Bonen teaches automated on-line flow analysis systems for determining bacterial endotoxin concentration in samples taken from production lines during the preparation of water, pharmaceutical products, and parenteral preparations. Bonen state that its automated system determines endotoxin concentration with good accuracy and reproducibility over the range of 0.01 to 10EU/mL (Bonen par. [0046-0047]). Bonen further expressly applies its endotoxin monitoring apparatus to water in a process loop such as a Water for Injection (WFI) or high-purity water system and teaches monitoring endotoxin using chromogenic or fluorogenic detection (Bonen par. [0055]).
Lorch further teaches continuous or real-time endotoxin monitoring of high-purity water, including water for injection. Lorch teaches that high-purity water includes water treated by reverse-osmosis, ultrafiltration, deionization, or distillation and expressly includes WFI and purified water within its contemplated pharmaceutical water systems. More particularly, Lorch teaches that the sensor may be operated with the sample continuously flowing past the sensing surface, wherein high-purity water flowing through a pipe is continuously withdrawn by a sampling system and caused to flow past the sensor, and the resulting sensor output changes as endotoxin in the sample bind to the sensing surface (Lorch par. [0072-0073]). Thus, Lorch teaches repeated/continuous measurement of endotoxin in the water sample at intervals necessarily encompassing measurement intervals of 30 minutes or less.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to modify Ase’s medical water production and endotoxin monitoring system to employ the fluorescence based LPS detection architecture taught by Cabral. Ase already teaches monitoring the quality of highly purified medical water, expressly including measuring endotoxin concentration, and separately teaches fluorescence-based analysis of contaminants in that water, while Cabral teaches fluorescent probes specifically capable of recognizing and detecting LPS through a fluorescent response. Applying Cabral’s known fluorescence-based LPS detection technique to Ase’s known water-quality monitoring system would therefore have constituted the predictable use of a known detection technique for its established purpose. It would have further been obvious to configure the resulting endotoxin detection system to quantify endotoxin over the 0.01-10 EU/mL range taught by Bonen because Bonen expressly teaches that range for automated endotoxin monitoring of WFI and high-purity-water production systems, thereby providing a known quantitative operating range directly applicable to the same type of water-monitoring environment. Finally, it would have been obvious to perform such endotoxin measurements repeatedly at intervals of 30 minutes or less in view of Lorch’s express teaching that continuous or real-time endotoxin monitoring is desirable in high-purity-water systems and its implementation in which water is continuously withdrawn from the process stream and passed over the endotoxin sensor. Configuring the combined system for measurements at intervals no greater than 30 minutes would therefore have been a predictable implementation of Lorch’s continuous-monitoring teaching, providing the known benefit of promptly detecting changes in endotoxin contamination during production.
Regarding claim 16, Ase in view of Cabral and Bonen and further in view of Lorch discloses or renders obvious a method of producing injection water, the method comprising: treating raw water by at least one of reverse osmosis filtration or ion exchange to obtain purified water (Ase Fig. 1, raw water lines 11-13 and RO device 5); obtaining injection water from said purified water by filtration via polymer membrane filtration or by distillation (Ase Fig. 1, ultrafiltration membrane device 7); and subjecting a test subject sample collected from the injection water to detection of an endotoxin (Ase p. 5 ET concentration measuring device), using a fluorescent substance that comprises a fluorescent site and a recognition site connected by a spacer (Cabral par. [0200] and claim 1, fluorescent sensor and linker), wherein: the recognition site recognizes a specific site of a molecular structure of said endotoxin (Cabral pars. [0009], [0091] and claim 41, recognition of negatively charged phosphate-containing membrane components including lipopolysaccharide); 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] linkers comprising short saturated alkyl chains); said detection of said endotoxin comprises quantifying an endotoxin concentration of from 0.01 EU/mL to 10 EU/mL (Bonen pars. [0046-0047]); and said detection of said endotoxin comprises repeatedly measuring said test subject sample at measurement intervals of 30 minutes or less (Lorch pars. [0072-0073] high-purity water continuously withdrawn from a process pipe and continuously flowed past an endotoxin sensor, with the sensor output responding as endotoxin binds).
Regarding claim 50, Ase in view of Cabral and Bonen and further in view of Lorch discloses or renders obvious the injection water production facility of claim 13, wherein said endotoxin detection device is configured to perform online analysis, offline analysis, or both (Bonen par. [0046]).
Regarding claim 52, Ase in view of Cabral and Bonen and further in view of Lorch discloses or renders obvious the method of producing purified water according to claim 15, wherein said detection of said endotoxin is performed by online analysis, offline analysis, or both (Bonen par. [0046]).
Claims 37, 18-19, 39 and 26-27 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 Bonen (US-20050124013-A1) as applied to claims 12 and 15 above, and further in view of Kasai "Staphylococcus aureus Detection by Fluorescent Silica Nanoparticles Modified with Metal Dipicolylamine Complexes".
Regarding claim 37, Ase in view of Cabral and further view of Bonen discloses the purified water production facility according to claim 12.
Ase in view of Cabral and further view of Bonen does not explicitly disclose wherein said recognition site comprises a dipicolylamino group to which a metal ion Mn+, wherein n is a natural number, is coordinated, and said specific site is a phosphate group of the endotoxin.
Kasai discloses fluorescent sensing compounds employing metal-dipicolylamine recognition sites for selective recognition of phosphate groups in water. Specifically, Kasai teaches fluorescent silica nanoparticles possessing metal-dipicolylamine binding sites with a coumarin fluorophore (M-dpa-HCC) for selective phosphate recognition in water (Kasai p. 749, left col. Fig. 1A, “M-dpa-HCC”). Kasai further teaches dpa-HCC having a dipicolylamino group coordinated with divalent metal ions, including Cu2+ and Zn2+ (Kasai p. 749, Fig. 1A and right col.), and further evaluates Ni2+-dpa-HCC complexes (Kasai p. 750). Kasai further teaches that the Cu-dpa-HCC complex functions by binding to phosphate moieties on a bacterial cell surface (Kasai p. 749, right col.). Thus, in view of Cabral’s teaching of fluorescent detection of lipopolysaccharide (LPS), a component of gram-negative bacterial membranes (Cabral, par. [0009]), Kasai discloses or renders obvious use of the recited metal-coordinated dipicolylamino recognition site for recognizing a phosphate group of the endotoxin.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to further modify the purified-water production facility of Ase in view of Cabral and Bonen to employ the metal-dipicolylamine phosphate-recognition chemistry taught by Kasai as the recognition site of the fluorescent substance. Cabral already teaches fluorescence-based detection of LPS using recognition of negatively charged phosphate-containing biological membrane components, while Kasai teaches that metal-coordinated dipicolylamine groups provide selective recognition of phosphate groups in water. A person of ordinary skill in the art therefore would have reason to employ Kasai’s known metal-dipicolylamine recognition chemistry in the previously combined fluorescence-based endotoxin detection system in order to provide a known recognition mechanism for selectively binding phosphate-containing sites of the endotoxin, with a reasonable expectation of success.
Regarding claim 18, Ase in view of Cabral and Bonen and further in view of Kasai discloses or renders obvious the purified water production facility according to claim 37, wherein said fluorescent substance is dpa-HCC represented by Formula 5 (Kasai p. 749 Fig. 1A M-dpa-HCC), having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site (Kasai p. 749 Fig. 1A depicting the hydroxycoumarin-carboxylic-acid fluorophore M-dpa-HCC), and wherein said spacer has one carbon atom (Kasai p. 749 Fig. 1A depicting the single methylene carbon connecting the coumarin fluorescent site to the dipicolylamino recognition site)
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Regarding claim 19, Ase in view of Cabral and Bonen and further in view of Kasai discloses or renders obvious 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 39, Ase in view of Cabral and Bonen and further in view of Kasai discloses or renders obvious the method of producing purified water of claim 15, wherein said recognition site comprises a dipicolylamino group to which a metal ion Mn+, wherein n is a natural number, is coordinated (Kasai p. 749, Fig. 1A and accompanying discussion, M-dpa-HCC having a metal-coordinated dipicolylamine group), and said specific site is a phosphate group of the endotoxin (Kasai p. 749, teaching selective phosphate recognition in water and binding of Cu-dpa-HCC to phosphate moieties on bacterial cell surfaces; Cabral par. [0009] teaching fluorescent detection of lipopolysaccharide (LPS) as a bacterial membrane component).
Regarding claim 26, Ase in view of Cabral and Bonen and further in view of Kasai discloses or renders obvious the method of producing purified water of claim 39, wherein the fluorescent substance is dpa-HCC represented by Formula 9 (Kasai p. 749 Fig. 1A M-dpa-HCC), having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site (Kasai p. 749 Fig. 1A depicting the hydroxycoumarin-carboxylic-acid fluorophore M-dpa-HCC), and wherein said spacer has one carbon atom (Kasai p. 749 Fig. 1A depicting the single methylene carbon connecting the coumarin fluorescent site to the dipicolylamino recognition site)
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Regarding claim 27, Ase in view of Cabral and Bonen and further in view of Kasai discloses or renders obvious the method of producing purified water of 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+).
Claims 38, 22, 23, 40 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 Bonen (US-20050124013-A1), and further in view of Lorch (US-20010040130-A1) as applied to claims 13 and 16 above, and further in view of Kasai "Staphylococcus aureus Detection by Fluorescent Silica Nanoparticles Modified with Metal Dipicolylamine Complexes".
Regarding claim 38, Ase in view of Cabral and Bonen and further in view of Lorch discloses or renders obvious the injection water production facility according to claim 13.
Ase in view of Cabral and further in view of Bonen and Lorch does not explicitly disclose the facility wherein said recognition site comprises a dipicolylamino group to which a metal ion Mn+, wherein n is a natural number, is coordinated, and said specific site is a phosphate group of the endotoxin.
Kasai discloses fluorescent sensing compounds employing metal-dipicolylamine recognition sites for selective recognition of phosphate groups in water. Specifically, Kasai teaches fluorescent silica nanoparticles possessing metal-dipicolylamine binding sites with a coumarin fluorophore (M-dpa-HCC) for selective phosphate recognition in water (Kasai p. 749, left col. Fig. 1A, “M-dpa-HCC”). Kasai further teaches dpa-HCC having a dipicolylamino group coordinated with divalent metal ions, including Cu2+ and Zn2+ (Kasai p. 749, Fig. 1A and right col.), and further evaluates Ni2+-dpa-HCC complexes (Kasai p. 750). Kasai further teaches that the Cu-dpa-HCC complex functions by binding to phosphate moieties on a bacterial cell surface (Kasai p. 749, right col.). Thus, in view of Cabral’s teaching of fluorescent detection of lipopolysaccharide (LPS), a bacterial membrane component (Cabral, par. [0009]), Kasai discloses or renders obvious use of the recited metal-coordinated dipicolylamino recognition site for recognizing a phosphate group of the endotoxin.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to further modify the injection water production facility of Ase in view of Cabral and further in view of Bonen and Lorch to employ the metal-dipicolylamine phosphate-recognition chemistry taught by Kasai as the recognition site of the fluorescent substance. Cabral already teaches fluorescence-based detection of LPS using recognition of negatively charged phosphate-containing biological membrane components, while Kasai teaches that metal-coordinated dipicolylamine groups provide selective recognition of phosphate groups in water. A person of ordinary skill in the art therefore would have reason to employ Kasai’s known metal-dipicolylamine recognition chemistry in the previously combined fluorescence-based endotoxin detection system in order to provide a known recognition mechanism for selectively binding phosphate-containing sites of the endotoxin, with a reasonable expectation of success.
Regarding claim 22, Ase in view of Cabral, Bonen and Lorch and further in view of Kasai discloses or renders obvious the injection water production facility according to claim 38, wherein the fluorescent substance is dpa-HCC represented by Formula 7 (Kasai p. 749 Fig. 1A M-dpa-HCC), having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site (Kasai p. 749 Fig. 1A depicting the hydroxycoumarin-carboxylic-acid fluorophore M-dpa-HCC), and wherein said spacer has one carbon atom (Kasai p. 749 Fig. 1A depicting the single methylene carbon connecting the coumarin fluorescent site to the dipicolylamino recognition site)
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Regarding claim 23, Ase in view of Cabral, Bonen and Lorch and further in view of Kasai discloses or renders obvious 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 40, Ase in view of Cabral, Bonen and Lorch and further in view of Kasai discloses or renders obvious the method of producing injection water according to claim 16, wherein said recognition site comprises a dipicolylamino group to which a metal ion Mn+, wherein n is a natural number, is coordinated (Kasai p. 749, Fig. 1A and accompanying discussion, M-dpa-HCC having a metal-coordinated dipicolylamine group), and said specific site is a phosphate group of the endotoxin (Kasai p. 749, teaching selective phosphate recognition in water and binding of Cu-dpa-HCC to phosphate moieties on bacterial cell surfaces; Cabral par. [0009] teaching fluorescent detection of lipopolysaccharide (LPS) as a bacterial membrane component).
Regarding claim 30, Ase in view of Cabral, Bonen and Lorch and further in view of Kasai discloses or renders obvious the method of producing injection water according to claim 40, wherein the fluorescent substance is dpa-HCC represented by Formula 11 (Kasai p. 749 Fig. 1A M-dpa-HCC) having 7-hydroxycoumarin-3-carboxylic acid as said fluorescent site (Kasai p. 749 Fig. 1A depicting the hydroxycoumarin-carboxylic-acid fluorophore M-dpa-HCC), and wherein the spacer has one carbon atom (Kasai p. 749 Fig. 1A depicting the single methylene carbon connecting the coumarin fluorescent site to the dipicolylamino recognition site)
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Regarding claim 31, Ase in view of Cabral, Bonen and Lorch and further in view of Kasai discloses or renders obvious 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 41, 33, 43 and 35 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 Bonen (US-20050124013-A1) as applied to claims 12 and 15 above, and further in view of Ozawa "Effect of Cyclodextrins on Saccharide Sensing Function of a Fluorescent Phenylboronic Acid in Water".
Regarding claim 41, Ase in view of Cabral and further in view of Bonen discloses the purified water production facility according to claim 12.
Ase in view of Cabral and further in view of Bonen does not explicitly disclose wherein said recognition site comprises phenylboronic acid, and said specific site is a sugar chain moiety of the endotoxin.
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 (Ozawa p.207 left col.). In view of Cabral’s teaching of fluorescent detection of lipopolysaccharide (LPS) as a bacterial membrane component (Cabral, claim 41 and par. [0009]), the phenylboronic-acid recognition chemistry taught by Ozawa discloses or renders obvious recognition of a sugar chain moiety of the endotoxin, because the saccharide portion of the lipopolysaccharide provides sugar/diol-containing sites of the type recognized by Ozawa’s phenylboronic-acid probes.
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 purified water production facility of Ase in view of Cabral and further in view of Bonen. Cabral already teaches fluorescence-based detection of lipopolysaccharide, while Ozawa teaches that phenylboronic-acid fluorescent probes recognize saccharide/diol moieties in water and produce a measurable fluorescence response upon saccharide binding. A person of ordinary skill in the art therefore would have had reason to employ Ozawa’s known phenylboronic-acid recognition chemistry in the previously combined fluorescence-based endotoxin detection system to provide a known recognition mechanism for binding the sugar-chain portion of lipopolysaccharide, thereby providing an alternative known fluorescent recognition chemistry for detecting endotoxin in an aqueous sample, with a reasonable expectation of success.
Regarding claim 33, Ase in view of Cabral and Bonen and further in view of Ozawa discloses the purified water production facility according to claim 41, wherein the fluorescent substance is C1-APB represented by Formula 13 (Ozawa p. 207 Fig. 1 “C1-APB”), having pyrene as said fluorescent site, and wherein said spacer has an amide bond (Ozawa p. 207 Fig. 1, C1-APB amide linkage)
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Regarding claim 43, Ase in view of Cabral and Bonen and further in view of Ozawa discloses the method of producing purified water according to claim 15, wherein said recognition site comprises phenylboronic acid (Ozawa p. 207 title and Fig. 1, fluorescent phenylboronic acid C1-APB), and said specific site is a sugar chain moiety of the endotoxin (Ozawa p. 207 teaching that phenylboronic acid forms stable cyclic esters with the diol moiety of sugars in water and that C1-APB functions as a fluorescent saccharide sensor; Cabral par. [0009], teaching lipopolysaccharide (LPS) as a bacterial membrane component).
Regarding claim 35, Ase in view of Cabral and Bonen and further in view of Ozawa discloses the method of producing purified water according to claim 43, wherein the fluorescent substance is C1-APB represented by Formula 15 (Ozawa p. 207 Fig. 1 “C1-APB”), having pyrene as said fluorescent site, and wherein said spacer has an amide bond (Ozawa p. 207 Fig. 1, C1-APB amide linkage)
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Claims 42, 34, 44 and 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 Bonen (US-20050124013-A1), and further in view of Lorch (US-20010040130-A1) as applied to claims 13 and 16 above, and further in view of Ozawa "Effect of Cyclodextrins on Saccharide Sensing Function of a Fluorescent Phenylboronic Acid in Water".
Regarding claim 42, Ase in view of Cabral and Bonen and further in view of Lorch discloses or renders obvious the injection water production facility according to claim 13.
Ase in view of Cabral and Bonen and further in view of Lorch does not explicitly disclose wherein said recognition site comprises phenylboronic acid, and said specific site is a sugar chain moiety of the endotoxin.
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 (Ozawa p.207 left col.). In view of Cabral’s teaching of fluorescent detection of lipopolysaccharide (LPS) as a bacterial membrane component (Cabral, claim 41 and par. [0009]), the phenylboronic-acid recognition chemistry taught by Ozawa discloses or renders obvious recognition of a sugar chain moiety of the endotoxin, because the saccharide portion of the lipopolysaccharide provides sugar/diol-containing sites of the type recognized by Ozawa’s phenylboronic-acid probes.
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 injection water production facility of Ase in view of Cabral and further in view of Bonen and Lorch. Cabral already teaches fluorescence-based detection of lipopolysaccharide, while Ozawa teaches that phenylboronic-acid fluorescent probes recognize saccharide/diol moieties in water and produce a measurable fluorescence response upon saccharide binding. A person of ordinary skill in the art therefore would have had reason to employ Ozawa’s known phenylboronic-acid recognition chemistry in the previously combined fluorescence-based endotoxin detection system to provide a known recognition mechanism for binding the sugar-chain portion of lipopolysaccharide, thereby providing an alternative known fluorescent recognition chemistry for detecting endotoxin in an aqueous sample, with a reasonable expectation of success.
Regarding claim 34, Ase in view of Cabral, Bonen and Lorch and further in view of Ozawa discloses or renders obvious the injection water production facility according to claim 42, wherein said fluorescent substance is C1-APB represented by Formula 14 (Ozawa p. 207 Fig. 1 “C1-APB”), having pyrene as said fluorescent site, and wherein said spacer has an amide bond (Ozawa p. 207 Fig. 1, C1-APB amide linkage)
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Regarding claim 44, Ase in view of Cabral, Bonen and Lorch and further in view of Ozawa discloses or renders obvious the method of producing injection water according to claim 16, wherein said recognition site comprises phenylboronic acid (Ozawa p. 207 title and Fig. 1, fluorescent phenylboronic acid C1-APB), and said specific site is a sugar chain moiety of the endotoxin (Ozawa p. 207 teaching that phenylboronic acid forms stable cyclic esters with the diol moiety of sugars in water and that C1-APB functions as a fluorescent saccharide sensor; Cabral par. [0009], teaching lipopolysaccharide (LPS) as a bacterial membrane component).
Regarding claim 36, Ase in view of Cabral, Bonen and Lorch and further in view of Ozawa discloses or renders obvious the method of producing injection water according to claim 44, wherein the fluorescent substance is C1-APB represented by Formula 16 (Ozawa p. 207 Fig. 1 “C1-APB”), having pyrene as said fluorescent site, and wherein said spacer has an amide bond (Ozawa p. 207 Fig. 1, C1-APB amide linkage)
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Claims 45 and 47 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 Bonen (US-20050124013-A1) as applied to claims 12 and 15 above, and further in view of Limbut "Capacitive biosensor for detection of endotoxin".
Regarding claim 45, Ase in view of Cabral and Bonen discloses or renders obvious the purified water production facility according to claim 12.
Ase in view of Cabral and Bonen does not explicitly disclose wherein said endotoxin detection device is configured to perform detection by flow injection analysis.
Limbut discloses detection of bacterial endotoxin using flow injection analysis. Specifically, Limbut teaches a flow-injection-based capacitive biosensor system for the direct detection and quantitative determination of bacterial endotoxin, wherein an endotoxin-containing sample is injected into a flowing buffer stream and passed through a biosensor flow cell for detection (Limbut p. 519, “Capacitance measurement”; p. 520, Fig. 1a, “flow-injection capacitive biosensor system”). Limbut further expressly analyzes endotoxin-containing samples using the flow-injection capacitive biosensor system and determines endotoxin concentration from a calibration curve generated using known endotoxin concentrations (Limbut p. 521, “Determination of the amount of endotoxin in real samples”). Thus, Limbut discloses an endotoxin detection device configured to perform detection by flow injection analysis.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to further modify the purified-water production facility of Ase in view of Cabral and Bonen to perform detection using the flow injection analysis arrangement taught by Limbut. The previously combined system already provides for collecting a water sample and quantitatively detecting endotoxin, while Limbut teaches flow injection analysis as a known sample-handling and analytical technique for direct quantitative endotoxin detection in which an endotoxin-containing sample is introduced into a flowing carrier stream and conveyed to a detector. A person of ordinary skill in the art therefore would have had reason to employ Limbut’s known flow-injection analysis arrangement for introducing and conveying samples through the previously combined fluorescence-based endotoxin detection system in order to provide rapid and efficient quantitative endotoxin analysis, with a reasonable expectation of success.
Regarding claim 47, Ase in view of Cabral and Bonen and further in view of Limbut discloses or renders obvious the method of producing purified water according to claim 15, wherein said detection of said endotoxin is performed by flow injection analysis (Limbut p. 520, Fig. 1a, “flow-injection capacitive biosensor system”).
Claims 46 and 48 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 Bonen (US-20050124013-A1), and further in view of Lorch (US-20010040130-A1) as applied to claims 13 and 16 above, and further in view of Limbut "Capacitive biosensor for detection of endotoxin".
Regarding claim 46, Ase in view of Cabral and Bonen and further in view of Lorch discloses or renders obvious the injection water production facility according to claim 13.
Ase in view of Cabral and Bonen and further in view of Lorch does not explicitly disclose wherein said endotoxin detection device is configured to perform detection by flow injection analysis.
Limbut discloses detection of bacterial endotoxin using flow injection analysis. Specifically, Limbut teaches a flow-injection-based capacitive biosensor system for the direct detection and quantitative determination of bacterial endotoxin, wherein an endotoxin-containing sample is injected into a flowing buffer stream and passed through a biosensor flow cell for detection (Limbut p. 519, “Capacitance measurement”; p. 520, Fig. 1a, “flow-injection capacitive biosensor system”). Limbut further expressly analyzes endotoxin-containing samples using the flow-injection capacitive biosensor system and determines endotoxin concentration from a calibration curve generated using known endotoxin concentrations (Limbut p. 521, “Determination of the amount of endotoxin in real samples”). Thus, Limbut discloses an endotoxin detection device configured to perform detection by flow injection analysis.
It would have been obvious to one of ordinary skill in the art prior to the effective filing date of the claimed invention to further modify the injection water production facility of Ase/Cabral/Bonen/Lorch to perform detection using the flow injection analysis arrangement taught by Limbut. The previously combined facility already provides for collecting a water sample and quantitatively detecting endotoxin, while Limbut teaches flow injection analysis as a known sample-handling and analytical technique for direct quantitative endotoxin detection in which an endotoxin-containing sample is introduced into a flowing carrier stream and conveyed to a detector. A person of ordinary skill in the art therefore would have had reason to employ Limbut’s known flow-injection analysis arrangement for introducing and conveying samples through the previously combined fluorescence-based endotoxin detection system in order to provide rapid and efficient quantitative endotoxin analysis, with a reasonable expectation of success.
Regarding claim 48, Ase in view of Cabral, Bonen and Lorch and further in view of Limbut discloses or renders obvious the method of producing injection water according to claim 16, wherein said detection of said endotoxin is performed by flow injection analysis (Limbut p. 520, Fig. 1a, “flow-injection capacitive biosensor system”).
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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/W.A.G./Examiner, Art Unit 1779
/Bobby Ramdhanie/Supervisory Patent Examiner, Art Unit 1779
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