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. DETAILED ACTION Application status Claims 1-29 are pending in this application. Priority The instant application is the 371 national stage entry of PCT/ NL2021/050167 , filed on 0 3 / 11 /20 21. Acknowledgment is made of applicant's claim for foreign priority under 35 U.S.C. 119(a)-(d) to a foreign patent application EPO EP20162431.9 filed on 03/11/2020 . Election Applicant's election with traverse of Group I, Claims 1- 12, 18-19 and 28-29 in the response filed on 01/20/2026 , is acknowledged. Applican ts argue that based on a Declaration under 37 C.F.R. § 1.130(a), wherein the co-inventor Giovanni Maglia addresses the Huang thesis (cited by the Office) filed on 01/20/2026 , the Huang thesis i s an exception under 35 U.S.C. § 102(b)(1)(A) such that it does not qualify as prior art since the Huang thesis was published less than one year before the effective filing date of the instant application. Thus, the claimed inventions have unity of invention, since they all share a special technical feature making a contribution over the prior art. Accordingly, Applicant respectfully requests withdrawal of the requirement for election. Applicants’ arguments have been fully considered but are not deemed persuasive for the following reasons. After reviewing the declaration under 37 CFR 1.130(a) by Giovanni Maglia filed on 01/20/2026 , t he Examiner notes that Huang’s thesis ("Engineering biological nanopores for proteomics study", thesis, University of Groningen, 28 November 2019 (2019-11-28), see IDS) as cited in the previous Restriction requirement mailed on 08/20/2025 falls under 35 USC 102(b)(1)(a) exception and does not qualif y as a prior art . However, in view of Lukoyanova et al. ( Conformational Changes during Pore Formation by the Perforin-Related Protein Pleurotolysin , PLOS Biology , 13(2): 16 Pages , published 02/05/2015, see IDS), which teach a high-resolution cryo-EM structure of wild-type PlyAB , which forms a barrel biological nanopore having a cylindrical trans chamber formed by (13 PlyB + 26 PlyA assembly) , an inner constriction with a diameter of 80 angstroms (= 8 nm ) , and a truncated-cone cis chamber (see Figures 1-5 and their related discussions) which meet the limitation s of claim s 1 -2 and 18-19 , and therefore, the shared technical feature of the groups is not a “special technical feature”, unity of invention between the groups does not exist. Claims 13-17 and 20-27 are withdrawn from further consideration by the Examiner, 37 CFR 1.142(b) as being drawn to a non-elected invention. For the reasons provided above, this restriction requirement is deemed proper, and therefore, it is made final. Information Disclosure Statement The information disclosure statement s (IDS) submitted on 09/08/2022, 12/20/2024, 03/19/2025 and 08/25/2025 are acknowledged . The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement s are being considered by the examiner. Claim Objections Claims 5-6 and 10-12 objected to because of the following informalities: Claims 5-6 and 10-12 recite the phrase “furthermore comprise” which can be substantially improved with respect to form. The Examiner suggests replacing the noted phrase with ---further comprise---. Appropriate correction is required. Objections to the Specification This application contains sequence disclosures that are encompassed by the definitions for nucleotide and/or amino acid sequences set forth in 37 CFR 1.821(a)(1) and (a)(2). However, this application fails to comply with the requirements of 37 CFR 1.821 through 1.825; Applicants’ attention is directed to the final rulemaking notice published at 55 FR 18230 (May 1, 1990), and 1114 OG 29 (May 15, 1990). To be in compliance, Applicants should identify nucleotide sequences of at least 10 nucleotides and amino acid sequences of at least 4 amino acids in the specification by a proper sequence identifier, i.e., “SEQ ID NO:” (see MPEP 2422.01). If these sequences have not been listed in the computer readable form and paper copy of the sequence listing, applicant must provide an initial computer readable form (CRF) copy of the “Sequence Listing”, an initial paper copy of the “Sequence Listing”, as well as an amendment directing its entry into the specification, and a statement that the content of the paper and CRF copies are the same and, where applicable, include no new matter as required by 37 C.F.R. 1.821(e) or 1.821(f) or 1.821(g) or 1.821(b) or 1.825(d). [1] See particularly Figures 2 and 3 of the specification containing amino acid /nucleic sequences, and therefore, those sequences should be represented by proper sequence identifier numbers. [2] The Examiner further notes that a sequencing listing paragraph is missing in the specification. The Examiner suggests inserting the following paragraph in the 1 st page of the specification with the relevant information filled in at ###: SEQUENCE LISTING A Sequence Listing is provided herewith as a Sequence Listing, " ### file name" created on ### date, and having a size of ### KB . The contents of the Sequence Listing are incorporated by reference herein in their entirety. [3] The substitution positions of PlyB disclosed in the specification do not match the numbering of amino acids as set forth in SEQ ID NO: 7, which is indicated to be the only SEQ ID NO disclosed which corresponds to PlyB . If the corresponding sequence of PlyB is not disclosed in the sequence listing as filed, Applicants must provide (1) an updated copy of the sequence listing containing the requisite sequences in computer readable form (CRF), (2) an amendment directing its entry into the specification, (3) a statement that no new matter has been added and (4) an amendment to the specification to identify each of the identified sequences by SEQ ID NO:, and (5) an incorporation by reference statement with the date of creation, sequence file name and size in bytes. – See also MPEP 2422. Appropriate correction is required. Claim Rejections - 35 U.S.C. § 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. Claims 1- 12, 1 8 -1 9 and 28-29 are rejected under 35 U.S.C. § 112 (b) , as being indefinite for failing to particularly point out and distinctly claim the subject matter which applicant regards as the invention. Claim 1 (2- 12, 17-18 and 28-29 dependent therefrom) recites the phrase “at least approximately” which indefinite . It is unclear because the term “approximately” encompasses a range which includes values which are higher and lower than the recited reference value. The term “at least” implies that only values equal to or higher than the recited reference value are encompassed. Therefore, in the absence of a clear definition of what is encompassed by the term “approximately”, the term "at least approximately" is unclear and confusing since the term refers to values which are equal or higher than undefined values which are either higher or lower than the recited reference value. In essence, the term eliminates the relevance of the recited reference point because the reference value becomes variable and undefined. The Examiner suggests replacing the noted phrase is ---at least---. Claims 3, 5-6, 8-12 and 28-29 recite substitutions, i.e., C62, C94, N153D/E, G265K/R, C487A/S/T, E306K/R, C62 and C94 are mutated to A, S, or T, etc , which are indefinite. The reason is that it is unclear where these substitutions are referring to in an amino acid sequence. The Examiner suggests inserting a corresponding SEQ ID NO which discloses the corresponding amino acid positions for the substitutions. Furthermore, the Examiner notes that the substitutions recited for PlyB do not match with SEQ ID NO: 7, which is the only amino acid sequence disclosed in the specification /sequence listing as being drawn to PlyB . In the interest of advancing prosecution, the noted phrase is interpreted broadly but reasonably as being ‘mutated’. Claim 3 recites the phrase “non- oxydizing residue” which is indefinite. The reason is that an amino acid can be oxidizing depending on the environment an amino acid residue is exposed to, and therefore, it is unclear which amino acid residues the phrase “non- oxydizing residue” is referring to since the instant specification does not clearly define ‘metes and bounds’ of what is encompassed by the noted phrase. The Examiner suggests replacing the noted phrase with the actual name of amino acid residues, i.e., serine, threonine or tyrosine. Claim 4 recites the phrase “increases the solubility of the subunit” which is indefinite. It is unclear what the ‘increase’ is in comparison to. The Examiner suggests replacing the noted phrase with the following, i.e., ---increases the solubility of the subunit compared to the PlyB subunits without said at least one mutation---. Claim 7 recites the phrase “increases the net positive charge” which is indefinite. It is unclear what the ‘increase’ is in comparison to. The Examiner suggests replacing the noted phrase with the following, i.e., ---increases the net positive charge of the inner surface of the nanopore compared to the inner surface of the nanopore without said at least one mutation---. The following is a quotation 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. Claims 1-12, 18-19 and 28-29 are rejected under 35 U.S.C. § 112(a), written description, 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(s), at the time the application was filed, had possession of the claimed invention. The instant claims are directed to a genus of beta -barrel biological nanopore s having a ny cylindrical trans chamber, an inner constriction with a diameter of at least approximately 2 nm, and a ny truncated-cone cis chamber . To satisfy the written description aspect of 35 U.S.C. § 112(a) for a claimed genus of [compositions or methods], it must be clear that: (1) the identifying characteristics of the claimed [compositions or methods] have been disclosed, e.g., structure, physical and/or chemical characteristics, functional characteristics when coupled with a known or disclosed correlation between function and structure, or a combination of these; and (2) a representative number of species within the genus must be disclosed. The Court of Appeals for the Federal Circuit has recently held that a “written description of an invention involving a chemical genus, like a description of a chemical species, ‘requires a precise definition, such as be structure, formula [or] chemical name,’ of the claimed subject matter sufficient to distinguish it from other materials.” University of California v. Eli Lilly and Co., 1997 U.S. App. LEXIS 18221, at *23, quoting Fiers v. Revel, 25 USPQ2d 1601, 1606 (Fed. Cir. 1993) (bracketed material in original). To fully describe a genus of genetic material, which is a chemical compound, applicants must (1) fully describe at least one species of the claimed genus sufficient to represent said genus whereby a skilled artisan, in view of the prior art, could predict the structure of other species encompassed by the claimed genus and (2) identify the common characteristics of the claimed molecules, e.g., structure, physical and/or chemical characteristics, functional characteristics when coupled with a known or disclosed correlation between function and structure, or a combination of these (paraphrased from Enzo Biochemical Inc. v. Gen-Probe Inc. (CAFC (2002) 63 USPQ2d 1609). The specification discloses only a single representative species of a nanopore comprising PlyA subunits as set forth in as set forth in SEQ ID NO: 5 with C62S and C94S mutations , and PlyB subu nits as set forth in SEQ ID NO:? with specific mutations N 72 D, N153D, G264R , A374T and A510V (note none of these mutations correspond to any SEQ ID NO disclosed in the sequence listing) (see Examples and pages 10 and 12 of the specification) . However, this single disclosed species fails to provide adequate written description for a genus of beta -barrel biological nanopore s having a ny cylindrical trans chamber, an inner constriction with a diameter of at least approximately 2 nm, and a ny truncated-cone cis chamber as encompassed by the claims, which encompasses any beta -barrel biological nanopore s made up of any polypeptide s having any sequence, optionally any PlyA and PlyB subunits having any amino acid sequence and mutations therein (italicized for added emphasis) . In this case, the specification fails to describe any identification of structural characteristics or properties of any polypeptides, optionally any PlyA /B subunits having any amino acid sequence with any mutations , such that they form any beta -barrel biological nanopore s with at least 2 nm pore, and which mutations introduced will lead to increased solubility or increased net positive charge as required in claims 4 and 7 . Taken together, the genus of “ any beta -barrel biological nanopore s made up of any polypeptides having any sequence, optionally with any mutations therein ” encompasses widely variant species, having essentially any structure. While M.P.E.P. section 2163 acknowledges that a single species can describe a genus, it also acknowledges that for a genus that encompasses widely variant species, disclosure of a single species within the genus fails to adequately describe all members of the genus. Please refer to the M.P.E.P. section 2163.05 [R-7.2022] under I, B for more details with respect to sufficient number of representative species that should be disclosed to describe a widely variant genus. Given the lack of additional representative species of the claimed genus which encompasses any beta -barrel biological nanopore s made up of any polypeptides having any sequence , optionally any PlyA and PlyB subunits having any amino acid sequence and mutations therein , Applicants have failed to sufficiently describe the claimed invention, in such full, clear, concise, and exact terms that a skilled artisan would recognize Applicants were in possession of the claimed invention. Applicant is referred to the revised guidelines concerning compliance with the written description requirement of U.S.C. 112(a) published in the Official Gazette and also available at www.uspto.gov . Claim Rejections - 35 USC § 102 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 – (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. Claim s 1-2 and 18-19 are rejected under 35 U.S.C. 102 (a)(1) as being anticipated by Lukoyanova et al. ( Conformational Changes during Pore Formation by the Perforin-Related Protein Pleurotolysin , PLOS Biology , 13(2): 16 Pages , published 02/05/2015, see IDS). The instant claims are drawn to a beta- barrel biological nanopore having a cylindrical trans chamber, an inner constriction with a diameter of at least approximately 2 nm, and a truncated-cone cis chamber . Lukoyanova et al. teach a high-resolution cryo-EM structure of wild-type PlyAB , which forms a barrel biological nanopore having a cylindrical trans chamber formed by (13 PlyB + 26 PlyA assembly) , an inner constriction with a diameter of 80 angstroms (= 8 nm ) , and a truncated-cone cis chamber , wherein a plurality of said nanopores are assembled into an amphipathic membrane (see Figures 1-5 and their related discussions), which anticipate Applicants’ claims 1-2 and 18-19. Therefore, teachings of Lukoyanova et al. anticipate the invention as claimed. Conclusion Claims 1-12, 18-19 and 28-29 are rejected for the reasons as stated above. Applicants must respond to the objections/rejections in this Office action to be fully responsive in prosecution. The instant Office action is non-final. Any inquiry concerning this communication or earlier communications from the examiner should be directed to JAE W LEE whose telephone number is (571)272-9949 . The examiner can normally be reached on M-F between 9:00-6:00. If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Manjunath Rao can be reached on ( 571)272-0939 . The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300. Information regarding the status of an application may be obtained from the Patent Application Information Retrieval (PAIR) system. Status information for published applications may be obtained from either Private PAIR or Public PAIR. Status information for unpublished applications is available through Private PAIR only. For more information about the PAIR system, see http://pair-direct.uspto.gov. Should you have questions on access to the Private PAIR system, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative or access to the automated information system, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000. /JAE W LEE/ Examiner, Art Unit 1656 /MANJUNATH N RAO/ Supervisory Patent Examiner, Art Unit 1656