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 Amendment
2. The amendment filed March 24, 2026 has been entered. Claims 1, 5-9, 11 and 14-15, have been entered. Claims 1-19 are under consideration in this Office Action.
Withdrawal of Claim Objections
3. The objection to claims 5-11 and 14-15 has been withdrawn in view of Applicants amendments.
Withdrawal of Claim Rejections
4. The rejection of claims 1-19 under 35 U.S.C. 102(a)(1) and/or (a)(2) as being anticipated by Raychaudhuri et al., is withdrawn in view of Applicants amendments and arguments.
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 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.
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.
5. Claims 1-19 are rejected under 35 U.S.C. 103 as being unpatentable over
Raychaudhuri et al., (US Pat Pub 20210046473 published 2021-02-18; priority to 2020-08-14) in view of Ciaurriz et al., (Beilstein J Nanotechnolo. 2017 Jan 25;8:244-253).
The claims are drawn to a Lyme disease multiplex enzyme linked immunosorbent assay ELISA method comprising: conducting a first immunoassay on a patient blood, serum, or plasma sample in which a primary antibody in the sample binds to at least one individual spot of an array of spots bound to a microtiter plate each containing a separate antigen of a plurality of antigens associated with Lyme disease, and a secondary antibody binds to human IgG; conducting a second immunoassay on the sample in which the primary antibody binds to at least one individual spot of an array of spots bound to a microtiter plate identical to that of the first assay, and a secondary antibody binds to human IgM; detecting a plurality of signals from the first immunoassay and second immunoassay using image auto-analysis, each signal corresponding to a spot in the first immunoassay or the second immunoassay; analyzing the plurality of signals using a machine learning (ML)-assisted algorithm to determine if the patient sample is positive for Lyme disease.
Raychaudhuri et al., describe multiplexed mass and nanoparticle detection imaging, tools, fluidics mad methods of using. Raychaudhuri et al., describe products and methods for profiling serum, e.g., blood serum, using a modified IRIS platform, wherein said modification relates to one or more of a fluidics pathway, a chip cartridge, a cassette housing for the chip cartridge and properly labeled gold nanoparticles observed in a wide field-of-view [para 25].
Raychaudhuri et al., describe obtaining step-wise information about the specimen status using a wide field-of-view modality is important as, for example, information first relating to IgM and then IgG gives significant diagnostic information. Also possible is the simultaneous injection of diluted biological samples (e.g., serum, plasma, whole blood, sputum, urine, pus) pre-mixed with labeled nanoparticles using a wide field-of-view modality[para 28]. Raychaudhuri et al., disclosed short assay times to test for specific binding of IgM and specific IgG signals, data for normal human serum and Lyme positive serum specimens may correlate to expected values obtained via ELISA. [para 75]. Thus teaching claim 1. The methods take advantage of the dual modality of direct mass detection and enhanced signal derived from traditional lateral flow immunoassay gold nanoparticle technology [para 6]. Raychaudhuri et al., relates to the direct detection of mass binding to the sensor surface and the subsequent detection of labeled gold nanoparticles. For example, target antigens, antibodies and nucleic acids are spotted down onto a functionalized IRIS chip using existing microarray technologies [para 44]. Gold nanoparticles are directly labeled with a ligand, e.g., such as an antibody targeting IgG or IgM, and these gold nanoparticles are then specifically associated with bound analyte [para 67]. The current multiplexed, array-based techniques may be utilized for detecting binding events (i.e., microarrays, microarray analyzers), but that many of these techniques require the use of fluorescently labeled antibodies and antigens [para 72]. The products and methods disclosed for short assay times, to test for mass binding, specific IgM and specific IgG signals. For example, in some embodiments, data for normal human serum and Lyme positive serum specimens may correlate to expected values obtained via ELISA [para 75]. The indirect IgM ELISA which incorporates these two specimens is also shown in Table 4. An approximate ELISA signal with the VoVo antigen for Lyme positive specimen was about an 8-fold over the normal human serum sample [para 90]. The ELISA tested for IgM reactivity to the VoVo antigen only, the IRIS system tested for reactivity to VoVo, VoBop, Vo4, BMSA1, BMN17 and included positive controls for human IgG and human IgM [para 91].
The assay detects sequentially useful information about the specimen (e.g., IgM antibody content followed by IgG antibody content) [para 31]. The use of disease specific analytes to detect direct mass accumulation followed by more specific step-wise conjugate nanoparticles to indicate disease status (e.g., anti-Human IgM gold nanoparticles and anti-Human IgG gold nanoparticles) for real-time and endpoint binding measurements [para 32]. Thus teaching claim 1. The disclosure comprises multiple distinct target compatible materials bound to a chip, wherein said materials target tick borne infections such as Lyme disease using a series of target analytes specific for Lyme infection including VoVo, VoBop, Vo4, OspC variants, DbpA, DbpB, VIsE, FlaB and C6 bound to the chip surface [page 38]. Thus teaching claims 5-7, 9 and 12-15. Any materials useful for spotting include but are not limited to DbpA (Lyme); DbpB (Lyme); BmpA (Lyme); VIsE (Lyme); FlaB (Lyme); VoVo (Lyme); VoBop (Lyme); Vo4 (Lyme); OCA (Lyme); OCB (Lyme); OCK (Lyme); and OCN (Lyme) [para 60]. Thus teachings 5-11.
The analysis method comprises analysis of binding events that incorporate dynamic or endpoint measurements; cut-off thresholds to determine positivity based upon one or multiple target binding events to a target; cut-off thresholds that incorporate one or multiple targets; and machine learning applied to categorizing specimens (e.g., positive or negative, IgM positive, antigen positive, acute, convalescent, etc.) based upon the real-time or endpoint measurements where a training set is used to train and properly classify specimens according to the desired status [para 39]. Thus teaching claim 4. Because the raw data is, itself, a series of sequentially collected images (an “image stack”), machine learning may be directly applied to either or both of the mass accumulation events and any of the subsequent gold nanoparticle binding events [para 4]. an apparatus comprising: an interferometric based imaging sensor (e.g., IRIS); multiple distinct target compatible materials bound to a functionalized chip surface (e.g., a microarray of spots using target analytes such as peptides, proteins and nucleic acids) [para 26]. Thus teaching the array of claims 18-19.
Binding signals are recorded at each individual spot by monitoring the intensity within the spot minus a nearby reference region as the control [para 59]. It is desirable to evaluate how diluted human serum binds to antigens that have been directly spotted onto the chip surface. To determine whether potential antibody binding (isotype independent) is significant and directly observable, five separate chips were spotted along with control analytes (including an anti-human IgM antibody). The results were observed, with IgM binding to the anti-human IgM capture antibodies is seen and negative control spots remain quiescent [para 63]. FIG. 3 illustrates the additional antibody “sandwiching” steps to determine antibody complementarity. The binding of molecules of interest in the samples to the spotted substrates on the chip and stored as data [para 80]. It is noted that there is about a 10-fold rise in signal at the anti-human IgG test spot [para 89]. An indirect IgM ELISA which incorporates these two specimens is shown in Table 4 [para 90]. An approximate ELISA signal with the VoVo antigen for Lyme positive specimen was about an 8-fold over the normal human serum sample [para 90]. Thus teaching claims 2-3 and 16-17. A similar 6-fold increase in signal is likewise observed at the end-point IgM values with the IRIS system as well as an 8-fold increase in signal at the end-point IgG values [para 90]. Thus teaching claims 16-17.
Example 1 shows Data sets have been obtained using the IRIS system applied to the detection Lyme antibodies in human serum [para 81]. Thus teaching claims 19-20. To evaluate human serum binding to Lyme antigen components, a series of Lyme target antigens were spotted on the chip: VoVo, VoBop and Vo4. The in-house Lyme antigen candidate has been the VoVo construct. Additional controls were included for anti-human IgM and anti-human IgG spots [para 81]. Thus teaching claim 7. An example normal human serum specimen binding to Lyme targets is shown in FIG. 7A and FIG. 7B and Lyme positive specimen binding to Lyme targets is shown in FIG. 8A and FIG. 8B. The highest climbing and second highest climbing lines on the charts show in each of FIG. 7A, FIG. 7B, FIG. 8A, and FIG. 8B represent the binding of the human IgM and IgG, respectively, to the appropriate capture antibodies on the test chip [para 83]. Table 1 shows positive control signals for the NHS and Lyme positive are indicated with the anti-human IgG and anti-human IgM capture values. Some specific mass accumulation is observed (above background/control values) with the Lyme positive specimen with the VoVo and BMSA1 (Babesia) constructs. See Fig. 2A showing dot platforms/plates.
The ELISA tested for IgM reactivity to the VoVo antigen only, the IRIS system tested for reactivity to VoVo, VoBop, Vo4, BMSA1, BMN17 and included positive controls for human IgG and human IgM. Mass binding and specific human IgM and IgG signals were also recorded [para 91]. For multiplex detection of targets, a multitude of disease substrates are spotted on the existing chip surface [para 74]. The IRIS that take advantage of the dual modality of direct mass detection in tandem with traditional lateral flow immunoassay gold nanoparticle technology [para 44]. The unique scattering properties of gold nanoparticles permit the digital detection of single virus and gold nanoparticles using the appropriate optical setup. Standard lateral flow immunoassay (LFI) gold conjugate techniques are used to specifically conjugate and functionalize nanoparticles, e.g., such as anti-human IgM and anti-human IgG [para 66]. Additionally, gold nanoparticle conjugates have been developed with lateral flow immunoassay techniques to create dried, room temperature stable conjugates. Due to the intrinsic light scattering nature of gold nanoparticles, there is also no photobleaching or ‘quenching’ of the signal that is often a concern of fluorescently labeled antibodies and antigens [para 73]. Thus, Raychaudhuri et al., teach ELISA multiplexed nanoparticle detection.
Ciaurriz et al., teach comparison of four functionalization methods of gold nanoparticles for enhancing the enzyme-linked immunosorbent assay ELISA. The enzyme-linked immunosorbent assay (ELISA) technique is based on the specific recognition ability of the molecular structure of an antigen (epitope) by an antibody and is likely the most important diagnostic technique used today in bioscience [abstract]. In this context, nanotechnology has offered a significant number of proposed solutions, mainly based on the functionalization of nanoparticles from gold to carbon which could be used as antibody carriers as well as reporter enzymes like peroxidase. Ciaurriz et al., teach use of gold nanoparticles (AuNPs) as a vehicle for secondary antibodies and peroxidase (HRP) [abstract]. The sensitivity increase of at least five times and a lower detection limit with respect to a standard ELISA of at least three times. Additionally, the assay time was remarkably decreased [abstract]. Ciaurriz et al., teach procedures that improve the detection limit, sensitivity or reduce the analysis time could have an important impact in several fields. In this respect, many methods have been developed for improving the technique, ranging from fluorescence substrates to methods for increasing the number of enzyme molecules involved in the detection such as the biotin–streptavidin method. In this context, nanotechnology has offered a significant number of proposed solutions, mainly based on the functionalization of nanoparticles from gold to carbon which could be used as antibody carriers as well as reporter enzymes like peroxidase. The IgG by AuNPs conjugated to goat anti-rabbit IgG (Ab) and HRP (AuNPs-Ab-HRP) was assayed to elucidate the best conditions for biomolecule binding and ELISA enhancement [Introduction]. Finally, AuNP functionalization methods an known for best enhancing the ELISA signal (Figure 1) [Results and Discussion].
Therefore, it would have been prima facie obvious at the time of applicants’ invention to apply Ciaurriz et al’s ELISA multiplexed nanoparticle detection to Raychaudhuri et al., describe multiplexed mass and nanoparticle detection immunoassay in order to provide the best conditions for biomolecule binding and ELISA enhancement. One of ordinary skill in the art would have a reasonable expectation of success by incorporating the multiplex ELISA techniques where the plate provides dried, room temperature stable conjugates and the gold nanoparticles, do not photobleach or ‘quench’ of the signal of the labeled antibodies and antigens.
Furthermore, no more than routine skill would have been required to incorporate the multiplex detection for Lyme disease using gold nanoparticles for enhancing the enzyme-linked immunosorbent assay ELISA having increased sensitivity, a lower detection limit and remarkably decreased assay time.
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 method of detecting Lyme disease using multiplexed ELISA, wherein the disease, IgM and IgG and there is no change in the respective function of the antibodies or reagents, 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. Thus, 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.
Response to Arguments
6. Applicant’s arguments, filed March 24, 2026, with respect to the rejection(s) of claims 1-19 under Raychaudhuri et al., have been fully considered and are persuasive. Therefore, the rejection has been withdrawn. However, upon further consideration, a new ground(s) of rejection is made in view of Raychaudhuri et al., in view of Ciaurriz et al’s ELISA multiplexed nanoparticle detection of antibodies.
Applicants argue that Raychaudhuri et al., describe the IRIS platform and a spotted chip. In this case, Applicants acknowledge that Raychaudhuri et al., an array of spots bound to a chip platforms. Raychaudhuri et al., teach a chip cartridge case and enclosure. The chip cartridge, a cassette housing for the chip cartridge and properly labeled gold nanoparticles observed in a wide field-of-view (i.e., not single nanoparticle counting) [para 25]. The multiple distinct target compatible materials bound to a chip [para 36]. There are multiple distinct target compatible materials bound to a chip, wherein said materials target a series of target analytes specific for Lyme infection including VoVo, VoBop, Vo4, OspC variants, DbpA, DbpB, VIsE, FlaB and C6 bound to the chip surface [para 38]. For example, target antigens and antibodies are spotted down onto a functionalized IRIS chip using existing microarray technologies [para 44]. Additionally, Raychaudhuri et al., clearly teach ELISA immunoassays. While Ciaurriz et al’s conjugates evaluated with a fixed concentration of IgG coated in a microplate well. Ciaurriz et al., multiwell plates were coated with a fixed concentration of rabbit IgG wherein the ELISA IgG probes uses gold nanoparticles conjugates, when both Raychaudhuri et al., and Ciaurriz et al., teach gold nanoparticles and antibodies. Furthermore, Ciaurriz et al., teach the advantages and benefits of employing ELISA techniques with an optimized approach, to provide increased sensitivity with reduced assay time.
Claim Rejections - 35 USC § 103
7. Claims 1-5, 9, 11, 13, and 15-19 under 35 U.S.C. 103 as being unpatentable over Juong et al., (ACS Nano 2020,14, 229-280) in view of Aucott et al., (US20160305956 published 2016-10-20, priority to 2015-04-16).
The claims are drawn to a Lyme disease multiplex enzyme linked immunosorbent assay method.
Juong et al., describe serodiagnostic test for early stage Lyme disease (LD) using a multiplexed paper based immunoassay and machine learning. Juong et al., created a cost-effective and rapid point-of-care (POC) test for early-stage LD that assays for antibodies specific to seven Borrelia antigens and a synthetic peptide in a paper-based multiplexed vertical flow assay (xVFA). We trained a deep-learning-based diagnostic algorithm to select an optimal subset of antigen/peptide targets and then blindly tested our xVFA using human samples (N(+) = 42, N(−) = 54), achieving an area-under-the-curve (AUC), sensitivity, and specificity of 0.950, 90.5%, and 87.0%, respectively, outperforming previous LD POC tests. With batch-specific standardization and threshold tuning, the specificity of our blind-testing performance improved to 96.3%, with an AUC and sensitivity of 0.963 and 85.7%, respectively [abstract]. The United States Center for Disease Control and Prevention (CDC) recommends a “two-tier” testing method, where the first-tier consists of a sensitive enzyme immunoassay (EIA) or immunofluorescence assay (IFA). If the first tier is positive or equivocal, a Western blot (WB) is then recommended for confirming the presence of 2 of 3 immunoglobulin M (IgM) antibodies and/or 5 of 10 immunoglobulin G (IgG) antibodies targeting Bb-associated antigens [page 230, col.1].
To overcome limitations, large-scale screening efforts alongside new epitope mapping and peptide synthesis are Juong et al., focused on developing a universal multiantigen detection panel, with, 5 to 10 LD-specific antigen targets being suggested for improving diagnostic performance for early LD. Juong et al., leveraged advances in computation and machine learning, to train powerful serodiagnostic algorithms with these rich, multiantigen measurements derived from well-characterized clinical samples, to ultimately outperform the traditional two-tier test. Deep learning, which refers to the use of artificial neural networks with multiple hidden layers, can be especially effective in developing nonlinear yet robust inference models from noisy data sets with complex and confounding variables [page 230, col. 2]. The cassette is divided into a top and bottom case, which can be separated through a twisting mechanism, revealing the multiplexed sensing membrane on the top layer of the case. The sensing membrane contains 13 immunoreaction spots defined by a black wax-printed barrier, where each spot is preloaded with a different capture antigen or antigen epitope-containing peptide as well as proteins serving as positive and negative controls to enable multiplexed sensing information. The bottom case is shown below with the sensing membrane containing the multiantigen panel, see Figure 1C. Figure 3B shows an antigen panel. Therefore, Joung et al., teach a kit or system comprising an array of Lyme antigens.
The sensing membrane contains 13 immunoreaction spots defined by a black wax-printed barrier, where each spot is preloaded with a different capture antigen or antigen epitope-containing peptide as well as proteins serving as positive and negative controls to enable multiplexed sensing information within a single test (Figure 1B) [page 232, col.1]. Thus teaching claim 3. The first top case facilitates the uniform flow of a serum sample, where LD-specific antibodies are bound to the detection antigens immobilized on the nitrocellulose surface. The second top case is then used for color signal generation, where a conjugate pad, releases embedded gold nanoparticles (AuNPs) conjugated to anti-human IgM or IgG antibodies. The AuNPs then bind to the LD-specific IgM or IgG antibodies previously captured on the sensing membrane, resulting in a color signal in response to the captured amount. After completion of these sandwich immunoreactions, the sensing membrane is immediately imaged by a custom-designed mobile phone reader (Figure 1D,E), which captures the background image (taken before the assay operation) and the signal image (taken after the assay operation) of the sensing membrane for subsequent analysis in a computer, where a neural network is used to ultimately determine the final result (seropositive or seronegative) [page 232, col.1]. Thus teaching claims 1 and 3-4. The antigen panel includes OspC, DbpB, BmpA Figure 2. Thus teaching claim 5, 9, 11, 13, 15 and 18-19. For the first tier, a combination of whole cell lysate enzyme linked immunosorbent assay (ELISA), C6 peptide EIA, or VlsE/ PepC10 ELISA testing was used. The second tier, comprised the standard IgM and IgG WB. Samples were considered seropositive if any of the three EIA tests in the first tier had a positive or equivocal (borderline) result and the second tier had a positive result for either the IgM or IgG WB as defined by the CDC recommendation (≥2 of 3 bands for IgM WB and ≥5 of 10 bands for IgG) [Clinical Study]. Thus teaching claim 2.
The entire assay operation takes 15 min (Figure 2A), and the assay reader, image processing, and neural network-based analysis are completed in under 30s (Figure 2B), as detailed in the Experimental Section. Figure 3. (A) Sensing membrane and map of the multiantigen panel. (B) Example images of IgM (left) and IgG (right) sensing membranes after activation by a human serum sample. (C) shows the multiantigen channel calculated over the training data set for the IgM (left) and IgG (right) sensing membranes ranked in descending order.
It is noted that claims 18-19 which are drawn to a kit and system only comprise an array of Lyme antigens. Although the reference does not specifically disclose that a kit would have instructions which teach how to use said kit, it would have been prima facie obvious to anyone of ordinary skill in the art to include instructions which describe how to perform the assay. Applicants should note that the printed matter on the instructions in a kit cannot serve to define the kit over the prior art. See In re Gulack 217 USPQ (CAFC 1983).
Aucott et al., teach a test sample can be detected simultaneously using a multiplex assay, such as a multiplex ELISA. Multiplex assays offer the advantages of high throughput, a small volume of sample being required, and the ability to detect different proteins across a board dynamic range of concentrations. An antibody can be bound to a solid substrate such as a well, such as commercially available plates. Each well can be coated with the antibody. The array may be used herein to refer to multiple arrays arranged on a surface, such as would be the case where a surface bore multiple copies of an array. Such surfaces bearing multiple arrays may also be referred to as multiple arrays or repeating arrays [para 54]. Kits for the detection can include pre-coated strip plates, biotinylated secondary antibody, standards, controls, buffers, streptavidin-horse radish peroxidise (HRP), tetramethyl benzidine (TMB), stop reagents, and detailed instructions for carrying out the tests including performing standards [para 68]. Multiplex arrays in several different formats based on the utilization of, for example, flow cytometry, chemiluminescence or electron-chemiluminesence technology, are well known in the art. The multiplex ELISA from Quansys Biosciences (Logan, Utah) coats multiple specific capture antibodies at multiple spots (one antibody at one spot) in the same well on a 96-well microtiter plate [para 78]. Chemiluminescence technology is then used to detect multiple biomarkers at the corresponding spots on the plate [para 78]. Example 1 is measured, using a multiplex-based approach, the levels of 58 immune mediators and 7 acute phase markers in sera derived from of a cohort of patients diagnosed with acute Lyme disease and matched controls. Serology results were determined following the CDC's two-tier testing algorithm measuring both IgM and IgG, with time of symptom onset being determined by a structured interview with the patient at the pre-treatment study visit [para 103 and 137].
The serology results were determined following the CDC's two-tier testing algorithm measuring both IgM and IgG, with time of symptom onset being determined by a structured interview with the patient at the pre-treatment study visit. The present invention contemplates traditional immunoassays including, for example, sandwich immunoassays including ELISA or fluorescence-based immunoassays, immunoblots, Western Blots (WB), as well as other enzyme immunoassays. Well-known mathematical methods for correlating a marker combination to a disease status employ methods like discriminant analysis (DA) Discriminant Functional Analysis (DFA), Tree-Based Methods, Generalized Linear Models, Generalized Additive Models, Fuzzy Logic based Methods, Neural Networks and Genetic Algorithms based Methods [para 103].
Therefore, it would have been prima facie obvious at the time of applicants’ invention to apply Aucott et al’s multiplexed ELISA detection to Juong et al., describe multiplexed mass and nanoparticle detection immunoassay in order to provide a multiplex assays offer the advantages of high throughput, a small volume of sample being required, and the ability to detect different proteins across a board dynamic range of concentrations. One of ordinary skill in the art would have a reasonable expectation of success by the multiplex ELISA techniques using titer plates and well known reagents where one can increase sensitivity or specificity of the diagnostic assay depending on the preference of the diagnostician. Furthermore, no more than routine skill would have been required to incorporate the multiplex detection for Lyme disease using multiplexed ELISA having increased sensitivity, which improves long term outcomes.
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 method of detecting Lyme disease using multiplexed ELISA, wherein the disease, IgM and IgG and there is no change in the respective function of the antibodies or reagents, 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. Thus, 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.
Response to Arguments
8. Applicant’s arguments, filed March 24, 2026, with respect to the rejection(s) of claims 1-5, 9, 11, 13,15-17 and 19 under Juong et al., have been fully considered and are persuasive. Therefore, the rejection has been withdrawn. However, upon further consideration, a new ground(s) of rejection is made in view of Juong et al., in view of Ciaurriz et al’s ELISA multiplexed nanoparticle detection of antibodies.
Applicants argue that Juong et al., a paper based multiantigen POC test which is multiplexed vertical flow assay instead of a multiplexed ELISA. In response to applicant's arguments against the Juong et al., reference individually, one cannot show nonobviousness by attacking references individually where the rejections are based on combinations of references. See In re Keller, 642 F.2d 413, 208 USPQ 871 (CCPA 1981); In re Merck & Co., 800 F.2d 1091, 231 USPQ 375 (Fed. Cir. 1986). In this case, it would have been prima facie obvious at the time of applicants’ invention to apply Aucott et al’s multiplexed ELISA detection to Juong et al., describe multiplexed mass and nanoparticle detection immunoassay in order to provide a multiplex assays offer the advantages of high throughput, a small volume of sample being required, and the ability to detect different proteins across a board dynamic range of concentrations.
Pertinent Art
9. The prior art made of record and not relied upon is considered pertinent to applicant’s disclosure.
See also, WO 2013067448, US 20190079084; WO 2019060885; US 20200255889, EP 3738970 and US 20210072257.
Initial evaluation of the Ibios Lyme Detect™ IgM IgG Elisa for
detection of Lyme disease. American Journal of Tropical Medicine and Hygiene, (2020) Vol. 103, No. 5. SUPPL, pp. 161. Abstract Number: 572.
Meeting Info: American Society of Tropical Medicine and Hygiene Annual
Meeting, ASTMH 2020. Virtual. 15 Nov 2020-19 Nov 2020.
INITIAL EVALUATION OF THE INBIOS LYME DETECT™ IGM IGG ELISA FOR DETECTION OF LYME DISEASE. Reyes, Dindo [Reprint Author]; Ireton, Gregory; Needham, James; Raychaudhuri, Syamal; Vallur, Aarthy C. InBios Int Inc, Seattle, WA USA. American Journal of Tropical Medicine and Hygiene, (NOV 2021) Vol. 105,
No. 5, Suppl. 5, pp. 315.
Ang et al., investigated the influence of assay choice on the results in a two-tier testing algorithm for the detection of anti-Borrelia antibodies. Eighty-nine serum samples from clinically well-defined patients were tested in eight different enzyme-linked immunosorbent assay (ELISA) systems based on whole-cell antigens, whole-cell antigens supplemented with VlsE and assays using exclusively recombinant proteins. A subset of samples was tested in five immunoblots: one whole-cell blot, one whole-cell blot supplemented with VlsE and three recombinant blots. The number of IgM- and/or IgG-positive ELISA results in the group of patients suspected of Borrelia infection ranged from 34 to 59%. The percentage of positives in cross-reactivity controls ranged from 0 to 38%. Ang et al., Eur J Clin Microbiol Infect Dis.2011 Jan 27; 30(8):1027-1032
Brandt et al., (Front. Public Health, 04 December 2019. Vol. 7), teach the evaluation of patient IgM and IgG reactivity against multiple antigens such as BBA69 and BBA73 together with antigens OspC, DbpA, FlaB, and VlsE in Stage 1 and Stage 2 early Lyme disease patient serum samples, and combined IgM and IgG responses in a multi-antigen approach for sensitivity and specificity determination of Lyme disease. The six antigen approach, whereby reactivity against at least 2 of 6 antigens constituted a positive serology, could increase sensitivity without compromising specificity.
US Patent 12014490 is directed to use of a rapid-test-validation computing device to determine if a result of a rapid test device is valid and identify the result. The rapid-test-validation computing device captures images of the rapid test device and employs a first artificial intelligence mechanism to determine if the rapid test device is properly aligned in the images. The rapid-test-validation computing device then employs a second artificial intelligence mechanism to determine if a result of the rapid test device is valid or invalid. If the result is valid, the rapid-test-validation computing device employs a third artificial intelligence mechanism to determine and present an objective output of the rapid test device result to a user; otherwise, the rapid-test-validation computing device presents a notification to the user that the rapid test device result is invalid. See also US Patent 12.249,407.
WO2015054319 Jewett et al., teach a single streamlined quantitative test that provides equivalent sensitivity and increased specificity compared to existing two-tier testing. iPCR combined with the DOC recombinant antigen only required testing of the IgG antibody fraction for a positive diagnosis and appears to have the potential to determine both the stage of Lyme disease. Serum samples from 16 healthy individuals were assayed by multiplex iPCR for both IgM (A) and IgG (B) host- generated antibodies against recombinant DbpA, BmpA, OspC, BBK19, OspA, RevA, Crasp2, and BBK50 antigen-coupled magnetic beads.
WO2011112805 Burbelo et al., teach methods, and kits for the diagnosis or detection of infection by a pathogen that causes Lyme disease in a subject.
Conclusion
10. No claims allowed.
11. Applicant's amendment necessitated the new ground(s) of rejection presented in this Office action. Accordingly, THIS ACTION IS MADE FINAL. See MPEP § 706.07(a). Applicant is reminded of the extension of time policy as set forth in 37 CFR 1.136(a).
A shortened statutory period for reply to this final action is set to expire THREE MONTHS from the mailing date of this action. In the event a first reply is filed within TWO MONTHS of the mailing date of this final action and the advisory action is not mailed until after the end of the THREE-MONTH shortened statutory period, then the shortened statutory period will expire on the date the advisory action is mailed, and any nonprovisional extension fee (37 CFR 1.17(a)) pursuant to 37 CFR 1.136(a) will be calculated from the mailing date of the advisory action. In no event, however, will the statutory period for reply expire later than SIX MONTHS from the mailing date of this final action.
12. 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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