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
1. Claims 1-8 are currently pending and under examination.
Information Disclosure Statement
2. The information disclosure statement (IDS) submitted on June 27, 2024 is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner. An initialed copy is attached hereto.
Specification
3. The disclosure is objected to because of the following informalities:
a) The cross-reference to related application section of the specification should be updated to show that the CON 17/502,320 has matriculated to US Patent 12,031,983.
b) Paragraph 0013 recites “.FIG.2” the first period needs to be removed.
Appropriate correction is required.
Claim Objections
4. Claim 4 is objected to because of the following informalities: On first sight, E. coli should be accompanied by Escherichia coli. Appropriate correction is required.
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.
5. Claim(s) 1-8 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Bui et al., Nano Lett., 2015; 15:6239-6246.
Independent claim 1 is drawn to a liquid composition comprising a mixture of streptavidin, gold nanoparticles (GNPs) probes comprising biotinylated and antibody modified gold nanoparticles (GNPs), and an amine-containing chemical loaded liposomes, wherein the composition is used to bind to a target analyte.
Independent claim 8 is drawn to a complex, wherein the complex comprises a target analyte moiety, a streptavidin moiety, a gold nanoparticles (GNPs) probe moiety comprising biotinylated and antibody modified gold nanoparticles (GNPs), and an amine-containing chemical loaded liposomes moiety, wherein the target analyte moiety is connected to the GNP probe moiety through a first antibody moiety on the GNP, the GNP moiety is further connected to the amine-containing chemical loaded liposome moiety through the streptavidin moiety, wherein the target analyte moiety is further connected with a second antibody that is pre-anchored on a lateral flow strip to ensure that the whole complex is anchored on the lateral flow strip.
Bui et al. disclose a plasmonic immunoassay with a binary (all-or-none) response. The presence of a single pathogen in the sample results in a chemical cascade reaction leading to a large red to dark-blue colorimetric shift visible to the naked eye. The immediate and amplified response is initiated by a triggered breakdown of cysteine-loaded nanoliposomes and subsequent aggregation of plasmonic gold nanoparticles (meets claim 1). The approach enabled visual detection of a single-digit live pathogen of Salmonella, Listeria, and E. coli O157 in samples (see abstract and page 6243; meets claims 2-4).
Figure 1. demonstrates a schematic of the liposome-amplified plasmonic immunoassay (LAPIA). One bacterium, molecule, or antigen can rapidly trigger a chemical cascade leading to a chromogenic aggregation of AuNPs. The reaction proceeds in different steps: (a) Capture of the target (biomarker, pathogen, toxin) using sandwich immunoassay. (b) After washing steps, biotinylated secondary antibody (polyclonal antiIgG) is allowed to interact with the immunocomplex. (c) After incubation and washing, streptavidin is added to interact and bind to biotinylated IgG. (d) After washing steps, biotin-conjugated liposomes containing cysteine are added to the medium followed by AuNP solution. (e) Addition of PBS-Tween-20 1× to the medium causes the breakdown of the liposomes and the release of cysteine, leading to immediate aggregation of gold nanoparticles and color shift from red to dark-blue (f) (see page 6240; meets claims 5-8).
Moreover, Bui discloses that three foodborne pathogens, Escherichia coli O157:H7, Salmonella typhimurium, and Listeria monocytogenes, were used as sample model (see page 6243). The preparation of Cys-liposome-biotin was performed using a mixture of L-α-phosphatidylcholine (PC), cholesterol, and phosphoethanolamine-conjugated biotin (PE-PEG2000-biotin) was dissolved in a chloroform solution (see page 6244; meets claims 7-8).
Since the Office does not have the facilities for examining and comparing applicants’ composition with the composition of the prior art, the burden is on applicant to show a novel or unobvious difference between the claimed product and the prior art. See In re Best, 562 F.2d 1252, 195 USPQ 430 (CCPA 1977) and In re Fitzgerald et al., 205 USPQ 594.
6. Claim(s) 1-8 are rejected under 35 U.S.C. 102(a)(1)/102(a)(2) as being anticipated by Abbas, US 2016/0252502 A1: Published: 9/1/16.
Independent claim 1 is drawn to a liquid composition comprising a mixture of streptavidin, gold nanoparticles (GNPs) probes comprising biotinylated and antibody modified gold nanoparticles (GNPs), and an amine-containing chemical loaded liposomes, wherein the composition is used to bind to a target analyte.
Independent claim 8 is drawn to a complex, wherein the complex comprises a target analyte moiety, a streptavidin moiety, a gold nanoparticles (GNPs) probe moiety comprising biotinylated and antibody modified gold nanoparticles (GNPs), and an amine-containing chemical loaded liposomes moiety, wherein the target analyte moiety is connected to the GNP probe moiety through a first antibody moiety on the GNP, the GNP moiety is further connected to the amine-containing chemical loaded liposome moiety through the streptavidin moiety, wherein the target analyte moiety is further connected with a second antibody that is pre-anchored on a lateral flow strip to ensure that the whole complex is anchored on the lateral flow strip.
Abbas discloses an immunoassay platform that can provide sensitive binary detection of a target such as, for example a molecule (e.g., a toxin, a contaminant, or other molecule of interest) or a pathogen (e.g., a bacterium, virus, parasite, etc.). Unlike a conventional ELISA immunoassay, where the color is generated by enzyme catalysis of a chromogenic substrate, the methods described herein use plasmonic colorimetry to generate a detectable signal that can be detected by the naked eye. The color change can be caused by nanoparticle aggregation. The assay may be performed without using an enzyme or, in some embodiments, the enzyme can be encapsulated in liposomes rather than bound to an antibody. The free enzyme provides better performance than an immobilized enzyme (e.g., in a conventional ELISA or plasmonic ELISA) (see paragraph 0040; meets claims 2-3, 7).
Exemplary applications include, for example, food safety, detection of food spoilage or adulteration, environmental testing, medical diagnosis, biodefense, and biological and medical imaging (e.g., immunohistology) (see paragraph 0041). The methods described herein can be used to confirm the absence of, for example, any pathogen (e.g., bacterium, virus, parasite, etc.) or molecule (e.g., toxin, contaminant, etc.) in food during processing or storage. The amplification system employed by the methods represents an extremely sensitive assay that can specifically detect the presence of a pathogen or molecule independent of its concentration. (see paragraph 0042; meets claims 3-4).
One exemplary embodiment of a lipolysis-enhanced nanoparticle aggregation immunoassay is shown as Assay 1 in FIG. 5. In the illustrated embodiment, a capture agent is immobilized to a substrate. The capture agent is selected to specifically bind to the target. The capture agent may be, for example, an antibody, or any other agent that specifically binds to the target to be detected (see paragraph 0045; meets claims 1, 7-8).
Moreover, Abbas discloses a composition that includes a recognition unit is contacted with the assay under conditions effective to allow the recognition unit to bind to the target captured by the capture agent. In some cases, the capture agent and the recognition unit may be related or the same, except that the recognition unit is functionally attached (e.g., conjugated) to a lysis agent. Thus, the recognition unit can include, for example, any antibody, aptamer, protein, single stranded DNA, receptor, or any other agent that specifically binds to the target to be detected. Unbound recognition unit is washed from the assay (see paragraph 0048; meets claim 1).
The composition includes liposomes. The liposome composition can also include a chromogenic reagent that produces a colorimetric signal in the presence of the chromogenic trigger. Alternatively, the chromogenic reagent can be provided in a separate composition and added to the assay before, at the same time as, or after the liposome composition is added (see paragraph 0050). In some embodiments, the chromogenic trigger can include, for example, cysteine, which can induce the aggregation of gold nanoparticles (AuNPs). In alternative embodiments, however, the chromogenic trigger can include, for example, any organic, inorganic, or hybrid agent that triggers a chromogenic reaction in specific conditions. Suitable agents include, for example, molecular crosslinkers (e.g., amino-thiolated molecules other than cysteine), pH indicators, acid/basic molecules, organic ions, and metal ions (see paragraph 0051; meets claim 1 and 5).
In some embodiments, the chromogenic reagent can include nanoparticles (e.g., gold nanoparticles) that aggregate in the presence of the chromogenic trigger (e.g., cysteine). In alternative embodiments, however, the chromogenic reagent can include any organic, metal, or hybrid molecule or particle that has the ability to generate color in certain conditions through a chromogenic reaction (see paragraph 0052; meets claims 1, 5-6).
Attachment can be direct (e.g., using a crosslinker between the liposome and the recognition unit) or indirect (e.g., by functionalizing the liposome and recognition unit with biotin and add streptavidin to assemble them) (see paragraph 0053; meets claims 7-8). The sample is contacted with the capture agent, washed, contacted with the recognition unit, and washed as described above. FIG. 5 (Assay 2) shows the assay in a generalized schematic, while FIG. 1 shows a particular embodiment in more detail. FIG. 1 shows the use of a recognition unit that includes an IgG that specifically binds to the target and a biotin-conjugated secondary anti-IgG antibody that binds to the IgG bound to the target. The liposome is conjugated to streptavidin, which specifically binds to the biotin, thus attaching the liposome to the recognition unit (see paragraph 0054; meets 1 and 6-8).
The sample is contacted with the capture agent, washed, contacted with the recognition unit, and washed as described above. FIG. 5 (Assay 3) shows the assay in a generalized schematic, while FIG. 1 shows a particular embodiment in more detail. FIG. 1 shows a recognition unit that includes a streptavidin-conjugated IgG that binds to the target that has been captured by the capture agent. A biotin-conjugated primary liposome is contacted with the assay under conditions effective to allow biotin-streptavidin affinity to attach the biotin-conjugated primary liposome to the streptavidin-conjugated IgG. Unbound biotin-conjugated primary liposome is washed from the assay (see paragraph 0057). The assay also involves a composition that includes secondary liposomes that contain the chromogenic trigger (e.g., cysteine, as shown in FIG. 2) and a composition that includes the chromogenic reagent (e.g., gold nanoparticles, AuNPs, as shown in FIG. 2) (see paragraph 0059).
In the embodiment illustrated in FIG. 2, the amplification possible is significant. Each primary liposome of, for example, 100 nm diameter can release approximately 1000 phospholipase molecules, each of which can lyse multiple secondary liposomes. Each secondary liposome can release billions of cysteine molecules, which induce aggregation of gold nanoparticles and the color change that can be visible to the naked eye. Thus, it is possible that an assay can accurately detect a single target bound to a capture agent. This detection can be further enhanced by using larger liposomes (200 nm or 400 nm) to contain more enzymes and more cysteine molecules (see paragraph 0061).
Lastly, Abbas discloses biotinylated cysteine-encapsulating liposomes were added to the particle solution (FIG. 9A-3). Gold nanoparticles and phosphate buffer containing Tween-20 were added. The Tween-20 breaks the liposomes attached to the nanoparticle through the microbe “bridge” and releases the cysteine payload (FIG. 9A-4). The free cysteine then induces the aggregation of gold nanoparticles, which yields a distinct red-to-blue color change (FIG. 9B). If the target microbe is not present in the system, then the antibodies and liposomes are simply washed away and the gold remains red upon addition of Tween-20 (see paragraph 0065).
Since the Office does not have the facilities for examining and comparing applicants’ composition with the composition of the prior art, the burden is on applicant to show a novel or unobvious difference between the claimed product and the prior art. See In re Best, 562 F.2d 1252, 195 USPQ 430 (CCPA 1977) and In re Fitzgerald et al., 205 USPQ 594.
Conclusion
7. No claim is allowed.
8. The prior art made of record and not relied upon is considered pertinent to applicant's disclosure. WO 2009/039502 A9.
9. Any inquiry concerning this communication or earlier communications from the examiner should be directed to LAKIA J JACKSON-TONGUE whose telephone number is (571)272-2921. The examiner can normally be reached Monday-Friday 930AM-530PM.
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, Anne Gussow can be reached at (571) 272-6047. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
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/LAKIA J JACKSON-TONGUE/Examiner, Art Unit 1645 June 11, 2026
/BRIAN GANGLE/Primary Examiner, Art Unit 1645
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