DETAILED ACTION
Notice of Pre-AIA or AIA Status
1. The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
Claim Amendments
2. The amendment filed May 19, 2025 has been entered. Claims 1, 5-10, and 20-23 have been amended. Claims 4 and 24-31 have been cancelled. Claims 1-3 and 5-23 are under consideration in this Office Action.
Information Disclosure Statement
3. The information disclosure statement (IDS) submitted on May 19, 2025 was filed. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
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 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.
4. Claims 1-3 and 5-23 are rejected under 35 U.S.C. 103 as being unpatentable over Alocilja et al., (US20140303012 published Oct. 2014; priority Aug 2012) and Ayed et al., (Phys. Chem. Chem. Phys. (2014) 16(39):21812-21819) in view of Lin et al., (Sensors and Actuators B: Chemical Volume 147, Issue 1, 18 May 2010, Pages 343-349) as evidenced by Hamer et al., ( Sci Rep. 2015; 5: 8716. Published online 2015 Mar 3).
The applied reference has a common inventor with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2) and 102(a)(1).
This rejection under 35 U.S.C. 103 might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C.102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B); or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement. See generally MPEP § 717.02.
The claims are drawn to a functionalized magnetic particle composition comprising (i) a magnetic particle core, and (ii) a biomimetic binding pair member bound to an external surface of the magnetic particle core, comprising a N-acetylgluosamine moiety and being capable of non-specific binding to a plurality of biological target analytes wherein the binding pair comprising the N-acetylglucosamine moiety comprises a pyrolysis by-product of chitosan and optionally one or more of an oligomeric unit of chitosan and a monomeric unit of chitosan.
Alocilja et al., disclose immobilized carbohydrate moieties on the biosensor provide a means for non-specific binding of a plurality of target analytes. When a sample containing the target analyte is applied or otherwise transported to the biosensor detection surface, non-specific binding interactions between the carbohydrate moiety and the analyte immobilize/retain the analyte at the detection surface [abstract].
The target analyte can comprise a bacterium, for example where (i) the bacterium is selected from the group consisting of Escherichia, Bacillus, Enterobacter, species of the foregoing genera, and strains of the foregoing; (ii) the specific binding pair member is capable of specifically binding to the selected bacterium; and (iii) the carbohydrate moiety is capable of non-specific binding to a plurality of bacteria from the group. Immobilizing the analyte conjugate can comprise contacting the analyte conjugate with the detection surface of the biosensor under conditions sufficient to non-specifically bind the carbohydrate moiety to the target analyte of the analyte conjugate [para 13]. Thus teaching claims 11-12. The carbohydrate moiety can be immobilized by reacting a glycoside containing the carbohydrate moiety and an amino group on the aglycone portion of the glycoside with an aldehyde functional group on the detection surface. As illustrated in the examples, surface aldehyde functional groups can be provided in the form of a glutaraldehyde layer applied to the detection surface. Reaction between the glycoside amino group and the aldehyde functional group forms an equilibrium-reversible imine bond, which can be further reduced to a stable secondary or tertiary amine covalent linking group (e.g., using cyanoborohydride) [para 37]; thereby teaching claim 10. Binding specificity (or specific binding) refers to the substantial recognition of a first molecule for a second molecule (i.e., the first and second members of the binding pair), for example a polypeptide and a polyclonal or monoclonal antibody, an antibody fragment (e.g., a Fv, single chain Fv, Fab′, or F(ab′)2 fragment) specific for the polypeptide, enzyme-substrate interactions, and polynucleotide hybridization interactions. The binding pair members exhibit a substantial degree of binding specificity and do not exhibit a substantial amount of non-specific binding (i.e., non-covalent binding between molecules that is relatively independent of the specific structures of the molecules, for example resulting from factors including electrostatic and hydrophobic interactions between molecules) [para 53]; thereby teaching claim 10.
The magnetic nanoparticle capture composition that includes a magnetic nanoparticle, and an additional specific binding pair member that is bound to the magnetic nanoparticle and is capable of specifically binding to the target analyte e.g., where the additional specific binding pair member can be the same or different as the binding pair member [para 0041]. The analyte probe and/or the analyte conjugate includes a magnetic or magnetically attractable moiety, for example as the separation/concentration component of the detection moiety), a magnetic field (e.g., using a magnet) can be applied to the sample to concentrate the analyte conjugate using an immunomagnetic separation process. Specifically, the applied magnetic field attracts the magnetic portion of the analyte conjugate, causing individual particles of the analyte conjugate to migrate to and concentrate in a region of the assay reaction vessel.
The magnetic nanoparticles are generally separable from solution with a conventional magnet. Suitable magnetic nanoparticles are provided as magnetic fluids or ferrofluids, and mainly include nano-sized iron oxide particles (Fe3O4 (magnetite) or γ-Fe2O3 (maghemite)) suspended in a carrier liquid. Such magnetic nanoparticles can be prepared by superparamagnetic iron oxide by precipitation of ferric and ferrous salts in the presence of sodium hydroxide and subsequent washing with water [para 46]; thus teaching claim 17. The final concentration of EAMNPs in each solution was 1.0 mg/ml. Immuno-conjugated EAMNPs (Mab-EAMNPs or BEAM nanoparticles) were stored at 4° C. Prior to experimental use, Mab-EAMNPs were either magnetically separated and concentrated or further diluted in 0.1 M PBS, in order to obtain solutions of Mab-EAMNPs at the following concentrations: 1.5 mg/ml, 1.0 mg/ml, 0.5 mg/ml, and 0.1 mg/ml EAMNPs [Example 1]; thus teaching claim 18.
The particulate compositions according to the disclosure generally include a conductive polymer bound to magnetic nanoparticles (e.g., a population of magnetic nanoparticles in which each nanoparticle generally has at least some conductive polymer bound thereto) [para 42]. Some conducting polymeric structures include polypyrrole [para 43]. The conductive polymers according to the disclosure are not particularly limited and generally include any polymer that is electrically conductive. Suitable examples of conductive polymers are polypyrroles. Other electrically conductive polymers include substituted and unsubstituted polypyrroles, polyfurans [para 44]. Thus teaching the pyrolysis by-products of chitosan. Suitable ferromagnetic nanoparticles include iron-containing magnetic metal oxides (paramagnetic or superparamagnetic), for example those including iron either as Fe(II), Fe(III), or a mixture of Fe(II)/Fe(III) [para 0047]; thereby teaching claims 2 and 20. The magnetic nanoparticles according to the disclosure are not particularly limited and generally include any nano-sized particles (e.g., about 1 nm to about 1000 nm) that can be magnetized with an external magnetic/electrical field [para 46]; thereby teaching claims 14 and 20. The Materials and Methods teach Ferric chloride hexahydrate, sodium acetate, sodium acrylate, sodium chloride (NaCl), ethylene lycol, ethylenediamine, hydrochloric acid, aniline, iron (III) oxide nanopowder, were used in the synthesis of the magnetic nanoparticles [para 71]; thus teaching claims 9 and 16. The magnetic nanoparticles and the monomer can be combined in any suitable weight ratio in the polymerization solution so that the resulting particulate composition has a desired balance of magnetic, electrical, and particle size properties. For example, the weight ratio of monomer:magnetic nanoparticles in the polymerization solution (or conductive polymer:magnetic nanoparticles in the resulting particulate composition) preferably ranges from about 0.01 to about 10, more preferably from about 0.1 to about 1 or about 0.4 to about 0.8, for example about 0.6 [para 48]; thus teaching claim 15. The carbohydrate moiety is suitably immobilized on the detection surface via a stable covalent bond. The covalent bond is stable against degradation or further reaction (e.g., via backward equilibrium reaction kinetics resulting in bond destruction and detachment of the carbohydrate moiety), in particular for extended periods [para 36]; thus teaching claim 13. The carbohydrate moiety can comprise a mannose moiety. More generally, the carbohydrate moiety can comprise at least one of a N-acetylgalactosamine moiety, a N-acetylglucosamine moiety (e.g., alone as a monosaccharide; together with the same or other saccharide moieties in an oligo- or polysaccharide). The carbohydrate moiety can be selected from the group consisting of monosaccharides, glycosides thereof, and combinations thereof [para 13]; thus teaching claims 3, 5 and 7.
The analyte can include pathogens of interest (e.g., bacterial pathogens such as E. coli O157:H7, B. anthracis, B. cereus, in addition to those listed above). The analyte also may be an antigen, an antibody, a ligand (i.e., an organic compound for which a receptor naturally exists or can be prepared [para 50]; thus teaching claims 11-12.
Ayad et al., teach a magnetic chitosan-polypyrrole-magnetite (Cs-PPypFe3O4) nanocomposite is prepared in a simple method via in situ chemical polymerization of pyrrole. Magnetic Fe3O4 nanoparticles of size in the range of 10-20 nm are successfully introduced into the Chitosan PPy matrix [Abstract]. Cs-PPypFe3O4) nanocomposite is easily separated from the reaction solution using an external magnet, which is very useful for practical applications [abstract]. PPy has triggered enormous research activities because of its fascinating features such as intrinsic high electrical conductivity and stability compared to the other conductive organic polymers [Introduction]. The magnetic iron oxide (e.g., maghemite (γ-Fe2O3) and magnetite (Fe3O4)) nanoparticles have recently gained increased interest due to their biocompatibility, strong super paramagnetic properties, and low toxicity. Furthermore, the magnetic nanoparticles demonstrate promising applications in drug delivery, cell separation, biosensors, and enzymatic assays. Introducing these nanoparticles into the proper matrix can lead to high adsorption performance and removal of contaminants from aqueous solutions with easy magnetic separation and recovery from the medium using an external magnet [Introduction]. Therefore, it is highly demanding to incorporate natural polymeric materials (e.g., chitosan (Cs)) into the PPy matrix, because such a feature is highly desirable for efficient adsorption. Several reports have been devoted to the use of Cs-based adsorbents as a promising material for the removal of heavy transition metals and several dyes. The combination of Cs and PPy is expected to improve the properties of both Cs and PPy especially the adsorption and the mechanical properties [Introduction]. The synergistic effects that combine the properties of PPy, Cs and uniformly dispersed Fe3O4 make the nanocomposite ideal for AG dye removal with large capacity, a fast adsorption rate, easy magnetic separation and recovery from the medium using a simple external magnet [Introduction]. The synergistic effect that combines the properties of PPy, Cs and uniformly distributed Fe3O4 nanoparticles makes the nanocomposite an ideal adsorbent and having a large adsorption capacity, a fast adsorption rate, and easy magnetic separation make the Cs–PPy–Fe3O4 nanocomposite for a highly promising adsorbent.
Lin et al., teach magnetoelastic-sensing device for the assay of pathogen with Escherichia coli O157:H7 (E. coli) as a target using chitosan-modified magnetic Fe3O4 nanoparticles (CMNPs) [abstract]. Thus teaching claims 2-3. Escherichia coli O157:H7 (E. coli) is an important enterohemorrhagic food-borne pathogen which causes hemorrhagic colitis [Introduction]; thereby teaching claims 11-12. The CMNPs were prepared by coating the Fe3O4 nanoparticles with a chitosan layer. E. coli driven CMNPs were magnetically adsorbed on the magnetoelastic (ME) sensor [Introduction].
The chitosan has a minimum of 90% deacetylation [Experimental], thus comprising a maximum of 10% acetylated units. Thereby teaching claim 24 and the acetylated and deacetylated units of claims 6, 7, 20, and 21.
Magnetic nanoparticles have received considerable attention for the diagnostic detection, separation, and imaging of pathogens. Nanoparticles of Fe3O4 are among the most widely used magnetic materials due to their chemical stability and ease of processing [Introduction]. Chitosan–Fe3O4 nano-biocomposite has been successfully used in immunoassay and immobilization applications [Introduction]. Magnetoelastic sensors of 20 mm length were cut from a thick ribbon of alloy (Fe40Ni38Mo4B18). Iron chloride hexahydrate, ferrous sulfate heptahydrate, ammonia, chitosan and glutaraldehyde, l-cysteine HCl (Cys) were purchased from commercial sources [Experimental]. Thus teaching claim 8. Chitosan is a cationic amine-rich polysaccharide, while E. coli is a gram-negative bacteria with an isotonic point of pH 4–5. Between pH 5–6.5 E. coli is negatively charged, while the CMNPs are positively charged. Therefore, the CMNPs can adsorb the negatively charged E. coli through electrostatic attraction, and then become adsorbed to the magnetoelastic sensor through magnetic flux resulting in an increased mass load on the sensor and corresponding decrease in resonance frequency (fr) [Section 3.2]; thus teaching claims 6-8. The described sensor can be used for the direct and selective detection of pathogens without time-consuming bacteria culturing [Section 3.4]. It is noted that while Lin et al., teach acetylated and deacetylated units of chitosan; Lin et al., does not evidence the oligomeric nature of chitosan.
Hamer et al., teach production of defined chitosan oligomers with a specific pattern of acetylation using a combination of chitin oligosaccharide deacetylases
Hamer et al., state chitosan is the completely or partially deacetylated form of chitin, thus being composed of GlcNAc as well as d-glucosamine (GlcN, d) units. Chitosan oligomers with specific pattern of acetylation (PA) can be produced by enzymatic deacetylation of chitin oligomers, but the diversity is limited by the low number of chitin deacetylases available [abstract]. Oligomers of chitin and chitosan show diverse biological activities [Introduction]. Chitosan oligomers have diverse biological activities with potentially valuable applications in fields like medicine. These properties may depend not only on the degrees of polymerization and acetylation, but also on a specific PA that cannot be controlled when the oligomers are produced by chemical hydrolysis [abstract]. These chitosans can be produced with different degrees of polymerization (DP) and different degrees of acetylation (DA), but their pattern of acetylation (PA) is invariably random. Hamer et al., started with chitin oligomers of defined degrees of polymerization (an), used the single enzymes for the in vitro generation of two different, fully defined mono-deacetylated chitosan oligomers (da(n-1) and ada(n-2)). As a proof-of-principle study, they combined the two deacetylases in a single reaction and generated the expected doubly deacetylated oligomers (dda(n-2)), showing that this approach opens a way to produce novel chitosan oligomers [Introduction]. Figure 2 shows non-reducing end generating da1–5 chitosan-oligomers and ada1–3 chitosan-oligomers. Thus deacetylated and acetylated chitosan oligomers and monomers are well known are producible.
Therefore, it would have been prima facie obvious at the time of applicants’ invention to apply the chitosan pyrolysis by-product, an chitosan-polypyrrole-magnetite nanocomposite as taught by Ayad et al., when Lin et al as evidenced by Hamer et al’s teach deacetylated and acetylated chitosan–Fe3O4 functionalized magnetic particle comprising a functionalized magnetic particle in order to provide stable low-cost chemical reagents adsorbed to the magnetic particle. One of ordinary skill in the art would have a reasonable expectation of success by incorporating aminated chitosan by product to the magnetic particle for separation because the functionalized particles have improved yield, chemical stability and easy rapid processing. Furthermore, no more than routine skill would have been required to incorporate the chitosan to target bacteria using deacetylated and acetylated chitosan oligomer-modified magnetic Fe3O4 nanoparticles.
Additionally, KSR International Co. v. Teleflex Inc., 127 S. Ct. 1727, 1741 (2007), discloses combining prior art elements according to known methods to yield predictable results, thus the combination is obvious unless its application is beyond that person's skill. KSR International Co. v. Teleflex Inc., 127 S. Ct. 1727, 1741 (2007) also discloses that "The combination of familiar element according to known methods is likely to be obvious when it does no more than yield predictable results". It is well known to take a, functionalized magnetic particle wherein there is no change in the respective function of the functionalized magnetic particles or binding pair member, thus the combination would have yielded a reasonable expectation of success along with predictable results to one of ordinary skill in the art at the time of the invention. Therefore, it would have been obvious to a person of ordinary skill in the art to combine prior art elements according to known methods that is ready for improvement to yield predictable results. The claimed invention is prima facie obvious in view of the teachings of the prior art, absent any convincing evidence to the contrary.
Pertinent Art
5. The prior art made of record and not relied upon is considered pertinent to applicant’s disclosure.
A chitosan aminated carbon-nitrogen magnetic nanoparticle is a specific type of chitosan-modified nanoparticle. To understand how these concepts are nested, it helps to break down the chemistry and terminology from the general category to this highly customized nanomaterial. See Piosik et al., (J. Phys. Chem. C (2022) 126 (42): 18100–18114). Lopez et al., (Int. J. Mol. Sci. 2013, 14(10), 19636-19650).
WO2016180973 Gartner et al., Pyrolytic method for the production of particles containing at least one metal species.
US Pat Pub 20170182188 Sabir et al., teach chitosan-coated magnetic nanoparticles for protein immobilization includes forming ferrous ferric oxide (Fe3O4) nanoparticles by co-precipitation and coating the nanoparticles with chitosan.
Conclusion
6. No claims allowed.
7. Any inquiry concerning this communication or earlier communications from the examiner should be directed to JA-NA A HINES whose telephone number is (571)272-0859. The examiner can normally be reached Monday thru Thursday.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor Peter Paras, can be reached on 571-272-4517. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/JANA A HINES/Primary Examiner, Art Unit 1645
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