CTNF 17/924,511 CTNF 101603 Notice of Pre-AIA or AIA Status 07-03-aia AIA 15-10-aia The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA. Priority Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. The instant application is a 371 of PCT/CN2021/092983 filed on 05/11/2021 which claims the benefit of CN202010390614.0 filed on 05/11/2020. No English translation of the certified copy of foreign priority was provided. Examiner obtained a machine translated version, verified priority, and attached it to this office action. All claims are examined here on their merits and are given the benefit CN202010390614.0 and are granted a filing date of 05/11/2020. Information Disclosure Statement The information disclosure statements (IDS) submitted on 12/09/2022, 06/12/2024, and 12/03/2024 are in compliance with the provisions of 37 CFR 1.97 with the exception of non-patent literature document CA on IDS 6/12/24 because the document provided does not include the cited pages. Accordingly, the information disclosure statements aside from non-patent literature document CA on IDS 6/12/24 are being considered by the examiner. 06-49-06 AIA The listing of references in the specification is not a proper information disclosure statement. 37 CFR 1.98(b) requires a list of all patents, publications, or other information submitted for consideration by the Office, and MPEP § 609.04(a) states, "the list may not be incorporated into the specification but must be submitted in a separate paper." Therefore, unless the reference has by cited by the examiner on form PTO-892, they have not been considered. Drawings 06-22 AIA The drawings are objected to because the difference between the data in Figure 2B and Figure 2C is not clear. Examiner recommends amending axis label for better clarity . Corrected drawing sheets in compliance with 37 CFR 1.121(d) are required in reply to the Office action to avoid abandonment of the application. Any amended replacement drawing sheet should include all of the figures appearing on the immediate prior version of the sheet, even if only one figure is being amended. The figure or figure number of an amended drawing should not be labeled as “amended.” If a drawing figure is to be canceled, the appropriate figure must be removed from the replacement sheet, and where necessary, the remaining figures must be renumbered and appropriate changes made to the brief description of the several views of the drawings for consistency. Additional replacement sheets may be necessary to show the renumbering of the remaining figures. Each drawing sheet submitted after the filing date of an application must be labeled in the top margin as either “Replacement Sheet” or “New Sheet” pursuant to 37 CFR 1.121(d). If the changes are not accepted by the examiner, the applicant will be notified and informed of any required corrective action in the next Office action. The objection to the drawings will not be held in abeyance. Claim Rejections - 35 USC § 112 07-30-02 AIA 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. 07-34-01 Claims 63, 65, 66, 69, 70, 72, 73, and 76 -79 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. Claims 63, 65, 69, 70, 73, and 76-79 use exemplary language, (i.e. “preferably”). It is unclear if applicant intends for these instances to limit the claim scope and therefore the metes and bounds of the claims are indefinite. Examiner will treat these elements as optional. Claims 61-63, 68, and 78 are indefinite because they are drawn to biological samples from a biological fluid, but the specification refers to a “peripheral body fluid sample” (pg. 2). Therefore, it is confusing whether the word sample in the claims refers to a bodily fluid or a component of the fluid, such as extracellular vesicles. For the purposes of examination, the word sample is treated as equivalent to extracellular vesicles. Claim 66 is indefinite because there are two potential interpretations of the claim. It is unclear whether claimed method is intended to indicate that the sample is from any of the listed cell types or whether the claimed method would specifically identify which of the listed cell types the sample is from. Claim 72 is rejected for double inclusion of the word “antibody” for two different functions. Amending the claim to a “first antibody” and a “second antibody” would overcome the rejection. Furthermore, claim 77 and claim 78 are rejected for including the word “reagent” for different functions. Amending the claims to include “first reagent”, “second reagent”, and “third reagent” would improve clarity. For the purposes of examination, the reagent that is specific for the central nervous system is the system-derived marker, and the reagent that is specific to the disease is the disease-associated marker. Claim 78 is rejected for lack of antecedent basis of “biological sample” in claim 77. 07-36 AIA The following is a quotation of 35 U.S.C. 112(d): (d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers. The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph: Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers. 07-36-01 AIA Claim s 73, 74, and 79 are rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends. Claim 73 is directed to a composition wherein the central nervous system (CNS) disease is selected from a list. This claim fails to further limit the claim from which it depends because it only recites intended use and does not change the composition itself. Claim 74 is directed to a composition wherein the central nervous system-derived markers are biomarkers derived from a list of neurons or cells. This claim fails to further limit the claim from which it depends because it only recites intended use and does not change the composition itself. Claim 79 is directed to a kit, wherein the central nervous system disease is selected from a list. This claim fails to further limit the claim from which it depends because it only recites intended use and does not change the kit itself . Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements. 07-30-01 AIA The following is a quotation of the first paragraph of 35 U.S.C. 112(a): (a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention. The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112: The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention. 07-31-02 Claim 66 is rejected under 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112 , first paragraph, as failing to comply with the enablement requirement. The claim(s) contains subject matter which was not described in the specification in such a way so as to enable one skilled in the art to which it pertains, or with which it is most nearly connected, to make and/or use the invention. Claim 66 recites a list of cells or neurons from which the CNS-derived marker could be derived. The claim could be interpreted to mean that the biomarker is from any of the recited cell types (ex. the sample has a broad neuronal origin) or it could be interpreted to mean that the biomarker is from one specific cell type only. The latter interpretation is not fully enabled, based on the state of the art at the time of filing. For example, L1CAM, which is one of the instant application’s claimed CNS-derived markers, was frequently used at the time of filing as a biomarker for isolating central nervous system extracellular vesicles because expression this protein is largely concentrated in CNS (Hornung pg. 11, line 1). However, the prior art does not use the L1CAM protein to specify what neuronal cell subtype the biological sample originated from (ex. GABAergic neurons, dopaminergic neurons, serotonergic neurons, etc.). Hornung also specifically states: “…isolation of microglial exosomes from blood currently is not possible” (pg. 12, right side, first paragraph). Lastly, Schwann cells are among the potential cell types for which the instant application claims the CNS-derived biomarker could originate from. However, Schwann cells are not found in the CNS and therefore cannot be CNS derived. Examiner notes that Hornung does teach an altered experimental protocol in which a biomarker of an astrocyte or oligodendrocyte origin could be identified. Except for these two cell types though, the specification, including the working examples, does not provide sufficient disclosure to supplement the state of the art at the time of filing such that one of ordinary skill in the art would be able enabled to identify which sub-cell type the biomarker is from. The quantity of experimentation needed to make or use the invention : The standard of an enabling disclosure is not the ability to make and test if the invention works but one of the ability to make and use with a reasonable expectation of success. A patent is granted for a completed invention, not the general suggestion of an idea (MPEP 2164.03 and Chiron Corp. v. Genentech Inc., 363 F.3d 1247, 1254, 70 USPQ2d 1321, 1325-26 (Fed. Cir. 2004). The instant specification is not enabling because one cannot follow the guidance presented therein, or within the art at the time of filing, and practice the claimed method without first making a substantial inventive contribution. Therefore, claim 66 is rejected under 35 U.S.C. 112, first paragraph, for failing to meet the enablement requirement. 07-31-03 AIA Claim s 60-62,64-66, 69, 72-74, and 76 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA), first paragraph, because the specification, while being enabling for detecting dual positive extracellular vesicles (i.e. Claims 63, 67, and 75) does not reasonably provide enablement for detecting dual positive samples generally . The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the invention commensurate in scope with these claims. The working examples only pertain to situations in which the samples are extracellular vesicles. Embodiments that include detection of a CNS-derived marker, but do not specify that the sample is an extracellular vesicle, could include the detection of a free marker which would not be beneficial in detecting a degenerative disease of the nervous system. Therefore, the scope of these claims is not fully enabled . 07-31-03 AIA Claim 62 is also rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA), first paragraph, because the specification, while being enabling for diagnosing a CNS disease based on the ratio of labeled versus total number of extracellular vesicles , does not reasonably provide enablement for diagnosing a CNS disease based on the total number of extracellular vesicles alone, which could be altered by other variables such as the volume of the fluid acquired . The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to use the invention commensurate in scope with these claims. The drawings and specification (pg. 35) specifically exemplify that the comparison between control and disease groups is based on the ratio of labeled to total number of extracellular vesicles, not on the total number alone. Therefore, the scope of this claim is not fully enabled . Claim 77 is rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention. The claim is drawn to a kit comprising reagents that can detect two different markers. There is nothing in the specification to define what a reagent means outside of its conventional definition. Therefore, any substance that can be used to detect the claimed markers is sufficient. In all of the working examples, the reagents are simply antibodies. The applicant has not provided adequate written description to claim every possible reagent that can detect a CNS marker and a CNS-disease marker. For detection of the desired markers, the present disclosure demonstrates that the applicant was in possession of antibody reagents only. Claim 78 rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention. The claim is drawn to a reagent and apparatus to acquire the biological sample. The specification discloses that the reagent and apparatus can include those commonly used by those skilled in the art. The working examples of both reagent and apparatus used for acquiring the biological sample have not provided sufficient breadth to show possession of every reagent and apparatus that could be used for acquiring a biological sample . Claim Rejections - 35 USC § 101 07-04-01 AIA 07-04 35 U.S.C. 101 reads as follows: Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title. Claims 60-70 are rejected under 35 U.S.C. 101 because the claimed invention is directed to a natural phenomenon judicial exception without significantly more. The claims recite the natural phenomenon whereby -a central nervous system disease is associated with the levels of system-derived markers (such as L1CAM or SHISA9) and disease-associated markers (such as Aβ1-40 (Aβ40) protein or tau protein) over control levels. This judicial exception is not integrated into a practical application because the method does not set forth any of the considerations outlined in MPEP 2106.05(a-c), (e) and (h). The additional steps do not constitute an improvement in technological field; and the steps recited in addition to the judicial exception do not integrate detection of the natural phenomenon into a particular treatment/prophylaxis according to MPEP 2106.04(d)(2). The claim(s) does/do not include additional elements that are sufficient to amount to significantly more than the judicial exception because the additional elements fail to provide either an inventive concept or impose meaningful limits upon the method such that the invention does not preempt every observance of the natural phenomenon itself. Claims 60-70 are directed to a method for diagnosis for a central nervous system disease in a subject comprising acquiring, pretreating, and detecting level and/or size of biomarker(s) in sample as compared to the level in a control sample. Methods are one of the statutory categories of invention (STEP 1:YES). Claims 60-70 recite diagnosis/prognosis/monitoring or comparing, all of which are abstract mental concepts that belong to enumerated group (c) of the Abstract Idea Groupings described in MPEP § 2106.04(a)(2): Mental processes — concepts performed in the human mind (including an observation, evaluation, judgment, opinion). But more importantly, the claims recite a natural correlation/phenomenon/law of nature whereby -condition, ex. Parkinson’s disease, pathology is correlated with changes in the level of samples with dual positive biomarker(s) in a biological fluid. Therefore, the claims recite at least one judicial exception (STEP 2A, Prong One: YES). According to Step 2A, Prong Two, set forth in set forth in MPEP 2106.04 II A (2), the claims are next evaluated with respect to whether the judicial exception is integrated into a practical application. These considerations are set forth in MPEP 2106.05 (a) through (c), (e), and (h). This analysis turns to the additional steps/elements recited within the claim. In independent claim 60 the additional steps/elements are: detecting a central nervous system-derived marker and a central nervous system disease-associated marker in a biological fluid, which is recited at with such a high level of generality that it encompasses any means known in the art (see MPEP 2106.05(e)). There are no additional elements that reflect an improvement within the technical field; there are no additional elements that apply the natural correlation/phenomena judicial exception to a particular treatment or which utilize a particular machine; there are no additional elements that effect a transformation; and, there are no additional elements that apply the judicial exception in some other meaningful way beyond generally linking it to a field, namely, central nervous system diseases. Dependent claims 63-70 do not include any additional steps that would amount to significantly more than the judicial exception recited in the claims from which they depend. Meanwhile, dependent claim 61 recites the additional steps of acquiring and pretreating the biological fluid, which is recited with such a high level of generality that it encompasses any means known in the art (see MPEP 2106.05(e)). Applicant admits in the specification that, “The method for acquiring the biological fluid of the subject may be any method known to those skilled in the art” (pg. 21, lines 5-10). In other words, the disclosure describes no new technology, but merely refers to techniques and elements that were known in the art at the time of filing. Furthermore, in the specification, the methods of pretreating the biological fluid may be: “centrifugation, ultracentrifugation, ultrafiltration tube filtration, polymeric sedimentation or specific antibody capture” but the applicant admits that these methods “all have the meaning understood by those skilled in the art, and those skilled in the art can select a suitable method according to extracellular vesicles to be acquired.” Therefore, in in accordance with MPEP 2106.07(a)(III)(A), the examiner has cited express statements in the specification indicating that the additional elements were sufficiently well-known that the specification does not need to describe the particulars thereof. In other words, the disclosure describes no new technology, but merely refers to techniques and elements that were known in the art at the time of filing. While dependent claim 62 recites the additional step of “counting” and/or “measuring”, this is recited with such a high level of generality that it encompasses any means known in the art (see MPEP 2106.05)). Hornung et al. Frontiers, 2020, hereafter Hornung, teaches that it was well established in the field of technical expertise to use immunocapture via antibody-coated beads and the use of nanoparticle tracking systems for detection of biological sample with CNS-derived biomarker (pg. 10, second paragraph, and Table 2). Furthermore, Lucien et al., Oncotarget, 2019 (hereafter Lucien) reveals that nanoscale flow cytometry is “specialized for high-throughput and multi-parametric analysis” of extracellular vesicles (pg. 1046, second paragraph), and according to Biggs et al., 2016, nanoscale flow cytometry was the only known method able to “provide quantitative information of multiple biomarkers expressed on EV’s” (pg. 8840, third paragraph). Therefore, the steps/elements recited in addition to the judicial exception (detecting, counting, and measuring both disease-associated and system-derived biomarkers from a biological sample) were all well understood, routine, conventional activities in the field of biomarker disease detection prior to filing the application at hand (STEP 2B: NO). The claimed steps/elements recited in addition to the judicial exception(s), alone or in combination, do not make an inventive contribution over the methods that were known in the art prior to filing, and they amount to mere observation of the natural phenomenon itself, by any means known, with the words “apply it” in order to append it to the field biomarker disease detection. Therefore, in accordance with MPEP 2106.07(a)(III)(C), the examiner has cited publications within the field of technical expertise, that demonstrate the well-understood, routine, conventional nature of the additional element(s). For all of these reasons, claims 60-62 are directed to the judicial exception without significantly more and are rejected. Claim Rejections - 35 USC § 102 07-06 AIA 15-10-15 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. 07-07-aia AIA 07-07 The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action: A person shall be entitled to a patent unless – 07-08-aia AIA (a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention. 07-15-aia AIA Claim(s) 72-75 is/are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by Hornung . Claims 72-75 read upon a composition comprising “an antibody, used for targeting a central nervous system-derived marker”; wherein the CNS-derived marker is selected from a list including L1CAM; and “an antibody, used for targeting a central nervous system disease-associated marker”; wherein the disease-associated marker is selected from a list including Aβ1-42. Claim 75 recites that the CNS-derived markers are located on the surface of extracellular vesicles. It is important to note that the recited elements of claims 73 and 74 are intended use and do not affect the composition itself; therefore, they do not need to be recited in the prior art to be anticipated. Hornung teaches an antibody, namely the anti-L1CAM antibody, used for targeting a central nervous system-derived marker L1CAM, and an antibody, namely a target specific antibody conjugate for Aβ1-42, used for targeting the central nervous system disease-associated marker Aβ1-42. Hornung also teaches that L1CAM is an adhesion protein and therefore is located on the surface on the extracellular vesicles (pg. 7, left side, third and fourth paragraph). Therefore, the claimed invention is anticipated by Hornung . Claim Rejections - 35 USC § 103 07-06 AIA 15-10-15 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. 07-20-aia AIA 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. 07-23-aia AIA 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. 07-20-02-aia AIA This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention. 07-21-aia AIA Claim s 60-72 and 76 is/are rejected under 35 U.S.C. 103 as being unpatentable over Hornung, as evidenced by Fiandaca et al. Alzheimer’s Dementia, 2015, in view of Lucien . Instant claims 60-71 are drawn to a method of diagnosing a central nervous system (CNS) disease from a biological fluid by counting and/or measuring the extracellular vesicles within the fluid that have both a CNS-derived biomarker and a CNS disease-associated biomarker. Instant claims 72 and 76 are drawn to a composition where the antibodies used for targeting the CNS-derived and CNS-disease associated markers are labeled antibodies. Hornung teaches acquiring a blood sample and pretreating it using centrifugation to isolate exosomes, which are a type of extracellular vesicle (pg. 8, right side, lines 20-30). Hornung further teaches immunocapture of the exosomes using antibody-coated beads to isolate exosomes with a CNS-derived marker, namely L1CAM, which is mainly expressed in the CNS. Hornung further teaches using a nanoparticle tracking system to measure exosome number and size (pg. 10, first and second paragraph, and Table 2). While Hornung does not explicitly teach that quantifying CNS disease-associated markers is accomplished using fluorescent labeling, it incorporates the techniques of Fiandaca by reference (pg. 7, left side, 18 lines from bottom). The methods of Fiandaca teach quantifying CNS disease-associated markers, including Aβ1-42, from the L1CAM positive exosomes using ELISA, through which the disease associated markers react with fluorescent labeled antibodies and are detected based on intensity of the labeled signal (pg. 3,4, Methods). Hornung further teaches that this method successfully diagnosed individuals with Alzheimer’s disease and even predicted its onset (pg. 7, left side, 12 lines from bottom). However, Hornung, as evidenced by Fiandaca, does not teach detection of the CNS-derived marker and CNS disease-associated marker at the same time . Hornung does not teach reacting the biological fluid with a labeled antibody and detecting the CNS-derived marker by measuring the intensity of the labeled signal, where the labeling could be one or more of fluorescence labeling, isotope labeling, enzyme labeling, chemiluminescence labeling, quantum dot labeling, and colloidal gold labeling. Lastly, Hornung does not teach counting and/or measuring samples in which both markers are detected, because the step at which Hornung teaches exosomes measurement occurs before the CNS disease-associated marker is detected. Lucien teaches using Apogee nanoparticle flow cytometry (pg. 1046, right side, middle paragraph) to simultaneously detect a disease related marker and a system specific marker on extracellular vesicles (pg. 1046, left side, 9 lines from bottom) using antibodies conjugated to fluorescent labels (pg. 1052, ride side first paragraph). Lucien further teaches counting extracellular vesicles in which both markers are present (pg. 1052, Table 3) as well as measuring their size, (pg. 1047, Table 3, Figure B). It would have been obvious to one of skill in the art to improve efficiency using flow cytometry to simultaneously detect multiple biomarkers on extracellular vesicles, since other traditional methods, as described in Hornung, could only do so in sequential steps (pg.7, right side, lines 17-20). In addition to its ability to simultaneously detect biomarkers, one would have been motivated to use flow cytometry because traditional methods of marker detection described in Hornung are time-consuming and tedious. Furthermore, using fluorescently labeled antibodies to detect the aforementioned markers would have been obvious, since that is a conventional implementation of flow cytometry. There would be a reasonable expectation of success when combining a successful method of simultaneous biomarker detection (nanoparticle flow cytometry), as described in Lucien, with biomarkers that successfully predict a CNS disease (ex. Alzheimer’s), as described in Hornung . 07-21-aia AIA Claim (s) 77-79 is/are rejected under 35 U.S.C. 103 as being unpatentable over Hornung, as evidenced by Fiandaca . Claims 77-79 are drawn to a detection kit comprising a reagent, used for detecting a central nervous system-derived marker in a biological fluid of a subject; wherein the possible CNS-markers are selected from a list including L1CAM; and a reagent, used for detecting a central nervous system disease-associated marker in the biological fluid of the subject; wherein the possible CNS-disease associated markers are selected from a list including Aβ1-42; wherein the CNS disease is selected from a list including degenerative diseases of the nervous system. The claims further comprise a reagent and apparatus for acquiring the biological sample of the subject. Regarding kit claims, patentability is determined as an assembly of interrelated parts and whether said assembly distinguishes over the prior art. Hornung teaches a reagent used for detecting a CNS-derived marker in a biological fluid, namely the anti-L1CAM antibody, and a reagent used for detecting a CNS disease-associated marker in a biological fluid, namely a target specific antibody conjugate for Aβ1-42, wherein the degenerative disease was Alzheimer’s. Hornung further teaches an apparatus, a centrifuge, for acquiring the biological sample (pg. 8, right side, lines 20-30). While Hornung does not explicitly teach a reagent for acquiring the biological sample, it incorporates the techniques of Fiandaca by reference. The methods of Fiandaca teach a reagent, Dulbecco’s balanced salt solution (DBS−2) with a protease inhibitor cocktail for acquiring the biological sample (pg. 4, line 1). It would have been obvious to package all of elements into a kit for commercial exploitation of the claimed invention. Therefore, claims 77-79 are rejected as obvious. Conclusion All claims are rejected. Any inquiry concerning this communication or earlier communications from the examiner should be directed to MICHELLE C BUCCINI whose telephone number is (571)272-1352. The examiner can normally be reached M-F 7:30-5 EST. Examiner interviews are available via telephone, in-person, and video conferencing using a USPTO supplied web-based collaboration tool. To schedule an interview, applicant is encouraged to use the USPTO Automated Interview Request (AIR) at http://www.uspto.gov/interviewpractice. If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Jeffrey Stucker can be reached at 5712720911. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300. Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. 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If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /MICHELLE CALLAHAN BUCCINI/ Examiner, Art Unit 1675 /JEFFREY STUCKER/ Supervisory Patent Examiner, Art Unit 1675 Application/Control Number: 17/924,511 Page 2 Art Unit: 1675 Application/Control Number: 17/924,511 Page 3 Art Unit: 1675 Application/Control Number: 17/924,511 Page 4 Art Unit: 1675 Application/Control Number: 17/924,511 Page 5 Art Unit: 1675 Application/Control Number: 17/924,511 Page 6 Art Unit: 1675 Application/Control Number: 17/924,511 Page 7 Art Unit: 1675 Application/Control Number: 17/924,511 Page 8 Art Unit: 1675 Application/Control Number: 17/924,511 Page 9 Art Unit: 1675 Application/Control Number: 17/924,511 Page 10 Art Unit: 1675 Application/Control Number: 17/924,511 Page 11 Art Unit: 1675 Application/Control Number: 17/924,511 Page 12 Art Unit: 1675 Application/Control Number: 17/924,511 Page 13 Art Unit: 1675 Application/Control Number: 17/924,511 Page 14 Art Unit: 1675 Application/Control Number: 17/924,511 Page 15 Art Unit: 1675 Application/Control Number: 17/924,511 Page 16 Art Unit: 1675 Application/Control Number: 17/924,511 Page 17 Art Unit: 1675 Application/Control Number: 17/924,511 Page 18 Art Unit: 1675