DETAILED ACTION
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 .
Priority
The present application was filed on 08/27/2023, which claims benefit of the foreign Application China 202211229850X, filed on 10/09/2022.
Claim status
Claims 1-13 are pending and examined.
Claim Objections
Claim 1 is objected to because of the following informalities: “capture antibodies as a test line and quality control antibodies as a control line are sprayed on the NC membrane”. The language of the claim does not make clear that capture antibodies are spray on the NC membrane to form a test line and quality control antibodies are spray on the NC membrane to form a control line.
Claim 2 is objected to because of the following informalities: “wherein the NIR-IL fluorescent rare earth nanoprobe test strip is a kind of RENPs with fluorescent emission peak in a NIR-II region”. Since the test strip cannot be a kind of RENPs, it appears that a NIR-IL fluorescent rare earth nanoprobe of the test strip is a kind of RENPs with fluorescent emission peak in a NIR-II region.
Claim 8 is objected to because of the following informalities: “one of a glass fiber and a polyester fiber” in lines 4-5 and 10. It should be read one of a glass fiber or a polyester fiber.
Claim 9 is objected to because of the following informalities: step 2 line 7, there should be an “a” before “microsphere washing liquid”.
Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claim(s) 2, 6, 7, 8, and 11 is/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.
Claim 2 recites “rare earth nanoprobe test strip is a kind of RENPs”. Claim 7 recites “rare earth nanoprobe test strip probe is a kind of carboxylated RENPs”. The phrase “a kind of” renders the claim indefinite because it is unclear what element can be used. Moreover, the claims 2 and 7 include elements not actually disclosed (those elements encompassed by “a kind of RENPs”), thereby rendering the scope of the claims unascertainable.
Claims 6-7 recites “the NIR-JI fluorescent rare earth nanoprobe test strip probe”. It is unclear if “nanoprobe” and “probe” refer to the same thing or the test strip comprises a nanoprobe and a probe.
Claim 6 recites the limitation "the NIR-LI fluorescent rare earth nanoprobe test strip probe" in lines 2-3. There is insufficient antecedent basis for this limitation in the claim because a probe of the test strip is not recited in claims 1-2, which claim 6 depends on.
Claim 6 recites a structure of the probe comprising NaYF4:X%Nd@NaYF4. The structure of the probe is not defined in the claim. The specification provides the definition for the elements forming the structure of the probe (see paragraphs 12-14) and the structure of the probe (see paragraph 50) but does not provide definition for the meaning of “:” and “@” in the structure, so it is unclear about the meaning of NaYF4:X%Nd@NaYF4. The claim is indefinite for failing to particularly point out and distinctly claim the subject matter.
Claim 8 line 9: The term “evenly” is a relative term which renders the claim indefinite. The term “evenly” is not defined by the claim, the specification does not provide a standard for ascertaining the requisite degree, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention.
Claim 11 recites the limitation “the microsphere washing solution” in line 2. There is insufficient antecedent basis for this limitation in the claim because a microsphere washing solution is not recited in claims 8-9, which claim 11 depends on. It is unclear that the microsphere washing liquid in claim 9 and the microsphere washing solution in claim 11 refer to the same thing. If the washing liquid and the washing solution in claims 9 and 11 refer to one liquid or solution, Applicant is suggested to select one term (liquid or solution) and add “a” before “microsphere washing” in claim 9 step 2 line 7 to avoid the antecedent problem.
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.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claim(s) 1-4 is/are rejected under 35 U.S.C. 102(a)(1) and (a)(2) as being anticipated by Yun et al. (CN111334282).
For claim 1, Yun teaches a rare earth detection kit, a detection card and microspheres thereof, as well as a preparation and detection method (see Abstract). The detection card comprises: a sample pad, a nitrocellulose (NC) membrane, an absorbent pad and a plastic backing (see Fig.1 and pages 2-3 and 6: teaching that the card comprises a sample pad, a nitrocellulose membrane, an absorbent paper, a plastic bottom card). The sample pad of detection card is also a conjugation pad because the end of the sample pad which is close to a coating film (i.e., nitrocellulose membrane) is sprayed with microsphere line 21 (i.e., detecting reagents) (see page 2 lines 35-40).
Yun teaches the sample pad 2, the conjugation pad 21, the NC membrane 3, and the absorbent pad 4 are superimposed on the plastic backing 1 successively along a horizontal direction (see Example 3 in page 5-6 and Fig.1).
Fig.1 of Yun CN111334282
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Yun teaches that detection antibodies labeled with rare earth nanoparticles (RENPs) are immobilized on the conjugation pad (see page 2 lines 35-40: teaching that a monoclonal antibody labeled with rare earth nano fluorescent microspheres is sprayed on the end of the sample pad, wherein the antibody is specific to a target analyte). The fluorescent microspheres of Yun are rare-earth fluoride near-infrared two-zone light-emitting nanomaterials (see page 2 lines 1-2, page 3 lines 55-60).
Yun teaches that capture antibodies as a test line 31 and quality control antibodies as a control line 32 are sprayed on the NC membrane (see page 2 lines 35-40, page 4 lines 35-36).
For claim 2, Yun teaches the test strip as claimed in claim 1, wherein the NIR-II fluorescent rare earth nanoprobe is RENPs with fluorescent emission peak in a NIR-II region (see page 2 lines 1-2: teaching that the rare earth nano-fluorescent microspheres emit fluorescence with a wavelength range of 1000-1100 nm; see page 3 lines 55-60: teaching that the fluorescent microspheres of Yun are rare-earth fluoride near-infrared two-zone light-emitting nanomaterials).
It is noted that the NIR-II with broadband emission spectra ranges from 1000 nm to 1700 nm (see paragraph 5 of the instant specification). Therefore, the teaching of Yun anticipates the limitation of claim 2.
For claim 3, Yun teaches the test strip as claimed in claim 2, wherein the RENPs are doped by one or more of rare earth ions (see page 1 lines 50-60: teaching that the rare earth nano fluorescent microspheres are doped by neodymium Nd3+).
For claim 4, Yun teaches the test strip as claimed in claim 3, wherein the RENP comprises an inner core and an outer shell (see page 1 lines 50-60: teaching that the RENP comprises an inner core NaGdxLu1-xF4:y Nd and an outer shell NaYF4).
Claim Rejections - 35 USC § 103
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.
Claim(s) 5-7 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yun et al. (CN111334282), as applied to claims 1-4, in view of Yang et al. (Recent advances in design of lanthanide-containing NIR-II luminescent nanoprobes, iScience, Volume 24, Issue 2, 19 February 2021, 102062).
For claim 5, Yun teaches the test strip as claimed in claim 4. Yun teaches that the structure of the nanoprobe is NaGdxLu1-y-xF4:yNd@NaYF4 (see page 1 lines 50-60). Yun teaches a surface of the nanoprobe is coated with carboxyl group to obtain water-soluble rare earth nano fluorescent microspheres (see page 2 lines 15-20). Yun does not teach the surface of the RENPs is modified with one or more of molecular polymers.
Yang teaches that lanthanide (Ln3+) containing nanoprobes are new generation of second near-infrared (NIR-II) probes (see Abstract and Introduction par.1), where lanthanide ions comprise Gd3+, Nd3+, Lu3+ (see page 2 last paragraph, see Table 1). From this teaching, it is noted that the nanoprobe of Yun is also Ln3+ containing nanoprobe because the nanoprobe of Yun contains Gd3+, Nd3+, and Lu3+.
Yang also teaches that organic polymers are the most frequently used agents for surface engineering of Ln3+ containing NIR-II nanoprobes, where organic polymers can comprise carboxyl group for creating hydrophilic surface (see page 13 par.2).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the surface of the nanoprobes taught by Yun, by coating the nanoprobe with polymers comprising carboxyl group as taught by Yang because Yun is generic to the nanoprobe coated with carboxyl group and Yang provides a species of nanoprobe coated with carboxyl group, which is the nanoprobe coated with polymer comprising carboxyl group. The nanoprobe of Yun is functionally equivalent to the nanoprobe of Yang in terms of creating a water-soluble nanoprobe. It is an obvious matter to try, namely choosing from a finite list of suitable, art recognized/known technique of coating nanoprobe with carboxyl group for obtaining a water-soluble nanoprobe.
One having an ordinary skill in the art would have had a reasonable expectation of success in modifying the nanoprobe of Yun to obtain water-soluble rare earth nano fluorescent microspheres by Yang’s technique because Yun and Yang are directed to the technique of creating the hydrophilic (also means water soluble) surface on Ln3+ containing nanoprobe, wherein the technique of creating the hydrophilic surface is coating carboxyl group on the surface of the nanoprobe, and wherein the nanoprobe is Ln3+ containing nanoprobe.
For claim 6, Yun teaches the test strip as claimed in claim 2, wherein a structure of the NIR-II fluorescent rare earth nanoprobe comprises: NaGdxLu1-y-xF4:yNd@NaYF4, where: NaGdxLu1-xF4 is the host or core, and the dopant ion is Nd3+; the colon ":" indicates neodymium doping; x and y are the molar percentage of rare earth ion doping, and the range of x is 20 %~90%, y ranges from 1%~10%; NaYF4 is the shell layer; @ means NaYF4 is coated on the surface of the core NaGdxLu1-y-xF4:yNd (see page 1 lines 55-60).
Compared to the claimed structure NaYF4: X%Nd@NaYF4, and X=I-100, the above teaching of Yun encompasses: the probe having the shell NaYF4, and the sodium tetrafluoride core (e.g., Na?F4 which ? is one or more doped ions, e.g., Gd or Lu or Nd), the nanoprobe is doped by x% ion Nd3+.
Yun teaches that the sodium fluoride core dopes ions Gd and Lu, which are members of lanthanide ion group. Yun does not specifically teach the sodium fluoride core comprising an inorganic matrix doped ion Y as claimed.
Yang teaches that Ln3+ containing NIR-II luminescent nanoprobes based on typical Ln3+ activators can comprise NaYF4:Nd3+@NaYF4 (see Table 1) which is read as a core NaYF4:Nd3+ surrounded by a shell NaYF4, and the core NaYF4-- is doped with rare earth Nd3+ which is one member of Ln3+ (see Fig.5, see page 1 par.1 and page 9 par.1). Yang also teaches the ion Nd3+ produces NIR-II light (see Abstract).
Yang supports that the core NaYF4 or NaGdF4 or NaLuF4 shares the same sodium tetrafluoride matrix, but the core is different from each other by the doped lanthanide ion (see Table 1: teaching a variety of Ln3+ containing NIR-II luminescent nanoprobes based on typical Ln3+ activators). Yang supports that the substitution of lanthanide ion in the core can be done by means of conventional techniques in the art (see Table 1).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the nanoprobe taught by Yun, by replacing the Gd and Lu ions in the core of the particle with the Y ion to arrive the claimed probe NaYF4:yNd3+@NaYF4, where y is the molar percentage of rare earth ion Nd3+ doping and y ranges from 1% to 10%. The obviousness is based on a simple substitution of one known element for another to obtain a functionally equivalent nanoprobe (e.g., the NIR-II fluorescent rare earth nanoprobe).
A skilled artisan would have had a reasonable expectation of success in modifying the nanoprobe taught by Yun because the modification would produce a functionally equivalent nanoprobe emitting NIR-II luminescent as taught by Yang. Therefore, the modified NIR-II luminescent nanoprobes would be successful for using in the test strip of Yun.
For claim 7, Yun and Yang teaches the test strip as claimed in claims 5-6. Yun and Yang teaches that the nanoprobe is a carboxylated RENPs. See discussion in claim 5.
Yun teaches wherein the NIR-II fluorescent rare earth nanoprobe test strip probe is carboxylated RENPs with a surface modified by one or more of sodium citrate, polyacrylic acid (PAA), distearoyl phosphoethanolamine-polyethylene glycol- carboxyl (DSEP-PEG-COOH), poly-dl-lactic-co-glycolic-polyethylene glycol- carboxyl (PLGA-PEG -COOHI), and polyethylene glycol-polylactic acid -carboxyl 12 (PEG-PLA-COOH) (see Yun page 2 lines 15-20: teaches the surface of the probe is modified with the carboxyl group; see Yang page 13 par.2: teaches the carboxyl group comprises DSPE-PEG2000-COOH).
Yun teaches that a particle size of the RENPs is in a range of 20 nm to 200 nm (see Yun page 1 lines 54-55: teaching that the particle size is 40nm~60nm).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use the specific carboxyl group DSPE-PEG2000-COOH taught by Yang to treat the surface of the probe taught by Yun in view of Yang because Yun is generic to the carboxyl group and Yang is specific for one carboxyl group composition that can be used to treat the nanoprobe’s surface. Therefore, the combination of Yun and Yang would have yield predictable result.
One having an ordinary skill in the art would have been motivated to use DSPE-PEG2000-COOH taught by Yang to treat the surface of the nanoprobe taught by Yun because the PEG2000-COOH treated nanoparticles can markedly improve the water solubility and biocompatibility of Ln3+-containing NIR-II luminescent nanoprobes (see Yang page 13 par.2).
Claim(s) 8 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yun, as applied to claim 1, in view of Merck (Rapid Lateral Flow Test Strips, Considerations for Product Development, 2013), Hao et al. (US 20160349251), and Han et al. (Low-cost, open-source 3D printed antibody dispenser for development and small-scale production of lateral flow assay strips, 2021).
For claim 8, Yun teaches the test strip as claimed in claim 1. Yun also teaches a preparation method of the NIR-Il fluorescent rare earth nanoprobe test strip comprising the following steps:
preparation of the sample pad: choosing one of a glass fiber or a polyester fiber, treating the one of the glass fiber or the polyester fiber with a sample pad treatment solution, and finally drying in an oven (see page 8 lines 41-43: teaching that sample pad is made of glass fiber; page 8 lines 20-30: teaching that a sample pad treatment solution is sprayed on one end of the sample pad, then the sample pad is placed in an oven and dry overnight);
preparation of the conjugation pad: diluting a RENPs labeling solution with a microsphere diluent to obtain a diluted RENPs labeling solution, evenly spreading the diluted RENPs labeling solution on the one of the glass fiber and the polyester fiber, and then drying in the oven (see page 8 lines 39-41: teaching that sample pad is made of glass fiber; page 8 lines 20-30: teaching that a diluted solution of microsphere diluent and rare earth nano-fluorescent microspheres labeled antibody in HCl solution is sprayed on the other end of the sample pad, then the sample pad is placed in an oven and dry overnight);
coating antibody of the NC membrane: preparing a capture antibody solution and a quality control antibody solution with a buffer, respectively; simultaneously spraying the capture antibody solution and the quality control antibody solution on the NC membrane to form the capture antibodies set as the test line (T line) and the quality control antibodies as the control line (C line) on the NC membrane, and then drying in the oven (see page 8 lines 30-35: teaching that a coating buffer (20mM pH8.0 Tris-HCl buffer containing 2.5% (w/w) sucrose) is used to adjust the concentration of PTH monoclonal antibody and goat anti-mouse IgG antibody to 1mg/ml, 1μl of coating volume of capture or control antibody buffer is used for a centimeter coating film, as the test line and quality control line, respectively, and dried in an oven; see page 8 lines 39-41: teaching that coating film is made of nitrocellulose);
preparation of the absorbent pad: cutting the absorbent pad to a target size (see page 8 line 40: teaching that an absorbent paper is placed on the backing (size 80*300mm) (the size is 28*300mm), which means the absorbent paper is cut to a target size);
assembling of a detection card: superimposing the sample pad, the conjugation pad, the NC membrane, the absorbent pad on the plastic backing successively along the horizontal direction (see Fig.1, Example 3 in page 5-6 and page 8 line 39-41); cutting the assembled pad and membrane into test strips, then putting the test strips into card slots to make detection cards, finally storing the detection cards in a dry environment (see page 6 lines 8-11, page 8 line 39-50)
Yun differs from the instant invention in the way of applying the treatment solution on the sample pad, which is soaking the pad in the solution, then shaking at room temperature (RT) for 2-3 hours (bold is the difference).
Yun differs from the instant invention in the way of applying the capture antibody and control antibody on nitrocellulose membrane, which is using a three-dimensional point spray platform (bold is the difference).
Yun does not teach using automatic cutting machine to cut the test paper board into test strips (bold is the difference).
Merck is generic about the method of producing a lateral test strip, where the sample pad can be impregnated with a variety of chemical agents designed to make the full range of samples compatible with the functioning of the test strip, and once impregnated with the treatment solution, the sample pads should be dried and stored at a certain condition (see page 27 Preparing Sample Pads section, see page 27 col.2 Storage section).
Merck also teaches using automatic cutting machine to cut the test paper board into test strips (see page 29 Slitting section).
Hao teaches a preparation and assemblage of the immunoassay test immunochromatographic strip (see par.94), which comprises submerging the sample pad of glass fiber with a treatment solution and then drying at 37° C. for 1.5 hours (see par.94).
Han teaches a 3D printed open-source antibody dispenser that can be easily built and used for the development of lateral flow assay (LFA) strips (see Abstract). Han demonstrates a uniform dispensing of capture antibody and control antibody to draw a test line and a control line on nitrocellulose membrane for lateral flow test strip (see Abstract, page 2 par.4).
Yun, Merck, and Hao do not teach the same method to treat the sample pad with the treating solution as claimed, which is soaking and shaking the sample pad in the treatment buffer for 2-3 hrs. at RT. However, Yun, Merck and Hao suggest a finite number of ways to prepare the sample pad for the test strip: Merck teaches that the sample pad is impregnated with the treatment solution; Yun teaches that the treatment solution is spraying to the sample pad; Hao teaches that the sample pad is submerged in the treatment solution. At the time of invention, it would have been obvious to one of ordinary skill in the art to try one or another way to apply the treatment solution on the sample pad and arrive the claimed method. A person of ordinary skill has good reason to pursue the known options within his or her technical grasp. The results of the combination are predictable (i.e., the sample pad is treated with the treatment buffer and is dried and stored at a certain condition) with a reasonable expectation of success.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use the automatic cutting machine to cut the test paper board into test strips taught by Merck in the preparation method of the NIR-II nanoprobe test strip taught by Yun. It is because Yun is generic about the cutting method and Merck provides one specific method to cut the board into strips. The method of Merck makes the cutting process quicker and more convenient.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to use 3D printed open-source antibody dispenser to draw a test line and a control line on the test strip of Yun because the dispenser can dispense capture antibodies and control antibodies uniformly as taught by Han.
Claim(s) 9-10 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yun, Merck, Hao and Han, as applied to claim 8, and further in view of Zhong et al. (In vivo molecular imaging for immunotherapy using ultra-bright near-infrared-IIb rare-earth nanoparticles, Nature Biotechnology volume 37, pages1322–1331 (2019)) and Delport et al. (Use of nano-particles in biosensing, Comm. Appl. Biol. Sci, Ghent University, 72/1, 2007).
For claim 9, Yun, Merck, Hao and Han teach the preparation method as claimed in claim 8. Yun also teaches a preparation of the RENPs labeling solution in the step 2 of claim 8 comprising:
activating treatment of RENPs: performing ultrasonic resuspension on carboxylated RENPs, and then discarding a supernatant after high-speed centrifugation to obtain a precipitate (see page 2 lines 20-25);
adding a 2-morpholinoethanesulphonic acid (MES) buffer into the precipitate, ultrasonically dispersing the precipitate added with the MES buffer, followed by adding an activator of 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide hydro (EDC) and a coupling agent of N- hydroxysulfosuccinimide (Sulfo-NHS), oscillating and performing a reaction to obtain a first product (see page 2 lines 20-25);
discarding a supernatant of the first product and obtaining a precipitate of the first product after high-speed centrifugation (see page 2 lines 20-25);
conjugation antibody of RENPs: adding the detection antibodies into the activated RENPs solution, oscillating for a reaction, then adding a bovine serum albumin (BSA) solution for sealing (see page 6 lines 19-25); discarding a supernatant of the second product and obtaining a precipitate of the second product after high-speed centrifugation, followed by adding microsphere washing liquid into the precipitate of the second product and ultrasonically suspending for 2-3 times; then adding a microsphere protective solution and storing at 4 °C for later use (see page 6 lines 19-25).
Yun does not clearly teach adding a phosphate buffer for ultrasonic dispersion to obtain an activated RENPs solution and adding an ethanolamine solution to terminate the reaction to obtain a second product.
Zhong teaches the preparation of the rare-earth lanthanide ion (Ln3+)-doped labeling solution (see page 1332). Zhong teaches that the carboxylated RENPs are kept in 1xPBS solution at 4 °C for long-term storage because the hydrophilic functionalized RENPs showed remarkable stability in aqueous buffers (1xPBS) and serum without aggregation and exhibited zero photo-bleaching (see page 1325 col.1 and page 1332 col.2).
Delport teaches techniques for the immobilization of different bio-molecules on nanoparticle NP (see Introduction). Delport teaches that for all protein bioconjugation experiments, EDC chemistry is applied for activating the carboxyl groups on NP before mixing protein with NP (see page 1 last paragraph). To prevent unspecific binding, an excess of ethanolamine is added after bio-molecule immobilization step to block the activated carboxyl groups on the surface of the NP, wherein ethanolamine binds all activated groups left on the surface and changes a charged carboxyl surface into a hydrophilic hydroxyl surface (see page 2 par.3).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the activating treatment step of Yun, by using a phosphate buffer for ultrasonic dispersion to obtain an activated RENPs solution as taught by Zhong because the hydrophilic functionalized RENPs showed remarkable stability in aqueous buffers (1xPBS) without aggregation and exhibited zero photo-bleaching (see Zhong page 1325 col.1 and page 1332 col.2).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the conjugation antibody to RENPs step of Yun, by using ethanolamine solution to terminate the conjugating reaction because Delport teaches that ethanolamine can block the activated carboxyl groups on the surface of the NP to prevent unspecific binding after the conjugation step is completed.
One having an ordinary skill in the art would have had a reasonable expectation of success in combining Yun, Zhong and Delport because they are directed to prepare nanoparticle probes for biosensing assay, where the nanoparticle probes need to be activated to conjugate with the biomolecule on their surface and the probes are used in immunoassay.
For claim 10, Yun, Merck, Hao, Han, Zhong, and Delport teach the preparation method as claimed in claim 9. Yun teaches activating the RENPs by incubating 200ul the carboxylated RENPs with 50 μl of 100 mg/ml EDC and 100 μl of 100 mg/ml Sulfo-NHS (see Yun page 8 lines 5-10).
Delport teaches that “to maximize the efficiency of the NP's, the conjugation chemistry and coating have to be optimized for every material, shape or size of the NP and every type of biomolecule” (see Introduction). Delport teaches activating the RENPs by incubating a quantity of NP with 0.15 mg/ml EDC (see page 1 last paragraph). To further improve the efficiency, the activated carboxyl groups were stabilized with NHS before reaction with the primary amines (see page 2 par.2). The optimal concentrations were found to be 12.5 mg/ml for both the EDC and NHS (see page 2 par.2).
Yun and Delport do not teach the same molar ratio of the carboxylated RENPs: the EDC: the Sulfo-NHS is 1: 5: 10 as claimed. However, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to define the optimal concentrations of each chemical used in the activating treatment step to maximize the efficiency of the NPs as taught by Delport. One having an ordinary skill in the art would have been aware that the varying amount of RENPs, EDC and Sulfo-NHS used for activating the carboxyl groups on the surface of NP taught by Yun and Delport is the consequence of the routine optimization process based on the material, shape or size of the NP and every type of biomolecule (see Delport Introduction). Absent unexpected results, it would have been obvious for one of ordinary skill to have arrived at the claimed molar ratio by routine optimization in order to uncover the optimum workable ranges of the method of preparing conjugated RENPs.
Claim(s) 11 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yun, Merck, Hao, Han, Zhong and Delport, as applied to claim 9, and as evidenced by Life Science network (10x PBS buffer (10x Phosphate Buffered Saline), 2013).
For claim 11, Yun, Merck, Han, Zhong, and Delport teach the preparation method in claim 9. Yun teaches the microsphere washing solution containing 0.5% BSA and 0.1% Tween-20 and the microsphere protective solution containing 0.5% BSA (see page 6 lines 20-25: teaching that a 0.5%BSA, 0.1% Tween-20 10~50mM, pH7.5~8.5 Tris-HCl preservation solution works as 2 solutions for washing and storing after conjugating antibody on RENPs).
Yun is silent to teach the solution contains a 10 millimoles per liter (mM) phosphate buffer saline (PBS) buffer.
On the other hand, Zhong teaches that the washing and storing solution after conjugating antibody on RENPs contains 1xPBS (see page 1332 col.2 par(s).1-3).
Life Science network teaches that 1xPBS is 10mM PBS buffer (i.e., 1x buffer will contain 137 mM NaCl, 2.7 mM KCl, 10 mM Na2HPO4, 2 mM K2HPO4).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the microsphere washing solution of Yun, using a 1x PBS because Zhong teaches the hydrophilic functionalized RENPs showing remarkable stability in aqueous buffers (1xPBS) without aggregation and exhibited zero photo-bleaching (see Zhong page 1325 col.1 and page 1332 col.2). Life Science network supports that the 1xPBS comprises 10mM PBS.
One having an ordinary skill in the art would have had a reasonable expectation of success in combining Yun and Zhong because they are directed to prepare nanoparticle probes for biosensing assay, where the nanoparticle probes need to be activated to conjugate with the biomolecule on their surface and the probes are used in immunoassay.
Claim(s) 12 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yun, Merck, Hao, Han, as applied to claim 8, and further in view of Trenkenschuh et al. (Freeze-thaw stability of aluminum oxide nanoparticles, International Journal of Pharmaceutics 606 (2021) 120932).
For claim 12, Yun, Merck, Hao and Han teach the preparation method in claim 8. Yun teaches wherein the microsphere diluent comprises a solution containing sucrose (see page 8 lines 24-25: teaching that a microsphere diluent comprises 20mM Tris containing 0.5% (w/w) BSA, 25% (w/w) sucrose).
Yun does not teach a 10 mM citric acid solution containing 1% sucrose
Merck teaches that the microsphere diluent comprises a solution containing 1%-10% sucrose which serves as a preservative and a re-solubilization agent (see page 24 Preparing conjugate pads).
Trenkenschuh teaches that NPs preserved in 10 mM Na-citrate buffer pH 5 and 8 (i.e., 10mM citric acid solution) is good, however, the size of NPs is still increasing in absence of further additives, e.g., sucrose (see page 8 col.1 par.3). The additives, e.g., sucrose improved NPs in preserved condition due to enhanced particle isolation and the formation of a stabilizing matrix (see page 8 col.1 par.3).
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the microsphere diluent of Yun, using a solution which comprises 10 mM citric acid solution containing 1% sucrose because Trenkenschuh teaches that the solution comprising 10 mM citric acid, and 1% sucrose improves the NPs isolation and stabilization. Moreover, the solution with 1% sucrose also serves as a preservative and a re-solubilization agent when applying the RENPs on the conjugation pad of the test strip as taught by Merck.
One having an ordinary skill in the art would have had a reasonable expectation of success in combining Yun, Merck and Trenkenschuh because they are directed to prepare nanoparticle probes for biosensing assay, where the nanoparticle probes are preserved for use later.
Claim(s) 13 is/are rejected under 35 U.S.C. 103 as being unpatentable over Yun, Merck, Hao, Han, as applied to claim 8, and further in view of Li et al. (Lateral Flow Immunochromatography Assay for Detection of Furosemide in Slimming Health Foods, Foods 2021, 10, 2041).
For claim 13, Yun, Merck, Hao and Han teach the preparation method in claim 8. Yun teaches wherein the sample pad treatment solution comprises a 20 mM tris(hydroxymethyl)aminomethane (Tris) buffer containing 0.5% surfactant, BSA (see page 8 lines 20-27: teaching that a solution comprises 20mM, pH8.0 Tris-HCl containing 0.5% S17 (i.e., surfactant), 0.1%BSA).
Yun does not teach the BSA used at 0.05%. Yun does not teach the Tris buffer further containing 0.05% Tween-20, 0.3% polyvinylpyrrolidone K30 (PVP-K30) and 0.05% PROCLIN-300.
Li teaches a preparation method of a lateral test strip (see pages 2-4). Li teaches the sample pad treatment solution comprising 0.5% BSA, 0.5% Tween-20, 0.3% PVP and 0.03% ProClin 300 (see page 4 par.2). Tween-20 is for a better release AuNPs–Abs probe and to adjust the chromatography speed, PVP is a steric stabilizer or capping agent to protect the AuNPs–Abs against agglomeration, and ProClin 300 is used as a preservative composition to prevent metamorphism (see page 3 par.3).
Li teaches that the sample pad plays a crucial role in reducing the interference of the sample matrix and affected the binding of the labeled probe on the nitrocellulose membrane, thereby affecting the color intensity and sensitivity of the test strip (see page 7 section 3.2.5). Li teaches an optimizing process to define the formula and the concentration of each component of the treating solution to achieve an optimal results for the nanoparticles lateral flow immunochromatography (see pages 5-7).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to modify the sample pad treatment solution of Yun, by adding Tween-20, polyvinylpyrrolidone K30 (PVP-K30) and PROCLIN-300 as taught by Li because Tween-20 is for a better release AuNPs–Abs probe and to adjust the chromatography speed, PVP is a steric stabilizer or capping agent to protect the AuNPs–Abs against agglomeration, and ProClin 300 is used as a preservative composition to prevent metamorphism (see Li page 3 par.3). One having an ordinary skill in the art would have been motivated to add Tween-20, polyvinylpyrrolidone K30 (PVP-K30) and PROCLIN-300 in the treating solution for obtaining a rapid, convenient and sensitive lateral flow immunochromatography (LFIA) based on nanoparticles (see Li in Abstract).
It would have been obvious for one of ordinary skill to discover the workable ranges of the concentration of each component of the treating solution by normal optimization procedures known in the art (e.g., taught by Li in pages 5-7) to achieve a rapid, convenient and sensitive lateral flow immunochromatography (LFIA) based on nanoparticles.
One having an ordinary skill in the art would have had a reasonable expectation of success in combining Yun and Li because they are directed to prepare sample pad for a lateral flow assay.
Conclusion
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/CHAU N.B. TRAN/ Examiner, Art Unit 1677
/BAO-THUY L NGUYEN/ Supervisory Patent Examiner, Art Unit 1677 January 26, 2026