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
This application has been filed on date 06/07/2022. Acknowledgment is made of the present application as a proper National Stage (371) entry of PCT/US20/64101, filed on 12/09/2020, which claims benefit of the 62/945,482, filed on 12/09/2019.
Claim status
Claim 4 is canceled. Claims 7-13 are withdrawn. Claims 1, 2, 3, 14, 15, 19, and 20 are amended, and claims 21-25 are new. Claims 1-3, 5-6 and 14-25 are examined.
Objections/Rejection status
The objection of claims 1-3, 14 and 20 is withdrawn in view of the amendment of the claims.
The rejection of claims 1-6 under 35 USC 112(a) scope of enablement is withdrawn in view of the amendment of the claims.
The rejection of claims 4-6 and 14-20 under 35 USC 112(b) is withdrawn in view of the amendment of the claims.
The rejection of claims 1-6 and 14-20 under 35 USC 101 is withdrawn in view of the amendment of the claim.
The objection of claim 2 is new.
Claim Objections
Claim 2 is objected to because of the following informalities:
It appears that a parenthesis is added in line 4 of claim 2 by mistake.
The required markers in the baseline, reference or control are not consistent with the markers in the test samples, e.g., “detecting in a plasma or serum sample of a patient with cancer elevated levels of PPIA, PDIA3 and at least one of … when compared with a baseline, reference or control levels of PPIA, EIF4H, PDIA3 and at least one of….” EIF4H is an optional marker in the test sample while in the control sample EIF4H is required to be tested together PPIA and PDIA3.
Appropriate correction is required.
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) 1 is/are rejected under 35 U.S.C. 103 as being unpatentable over Thalin (WO2017027379) in view of Wang et al. (Platelet-derived ERp57 mediates platelet incorporation into a growing thrombus by regulation of the αIIbβ3 integrin, Blood (2013) 122 (22): 3642–3650), Nigro et al. (Cyclophilin A: a key player for human disease, Cell Death and Disease (2013) 4, e888) Brown (US20140220580), and Mosawy (Effect of the Flavonol Quercetin on Human Platelet Function: A Review, Food and Public Health 2015, 5(1): 1-9).
For claim 1, Thalin discloses a method for diagnosing cancer or cancer-associated thrombosis (see Title, see page 1 lines 7-10). The method comprises detecting in a plasma sample of individual a presence or amount of a biomarker, and indicating cancer-associated thrombosis or an elevated risk for cancer associated thrombosis (see page 5 lines 22-33). The method further comprises comparing the plasma levels of the biomarker in cancer patient with that in healthy individual (see page 19 lines 21-31), wherein the cancer patients are from lung cancer, pancreatic cancer, colorectal cancer (see page 38 lines 15-31).
The method further comprises a step of administering a treatment to the thrombosis patient, e.g., antiplatelet agents (see page 43 lines 5-11, 22-31, and page 44 lines 1-18).
The cancer patient in the method is actively undergoing treatment for cancer, the treatment comprising chemotherapeutic treatment and/or radiation, and/or immunotherapy, and/or cell therapy (see page 6 lines 22-33 and page 7 lines 1-11: teaching a method for screening for thrombosis or an elevated risk for thrombosis in cancer individuals following chemotherapy to identify whether an individual with cancer that has received chemotherapy has thrombosis).
Thalin does not teach that the biomarkers are PPIA, PDIA3 and at least one of EIF5A, EIF4H, EIF4a3, UBE2N, UBE2L3, UBE2I, and HSP70.
While Thalin teaches treating thrombosis patients by using an antiplatelet agent, Thalin does not use an effective amount of rutin, or quercetin, or rutin derivative compound, or quercetin derivative compound for treatment.
Wang teaches that platelet-derived ERp57 plays an important role in physiologic platelet function and thrombosis (see Abstract). In particular, the increasing surface expression of ERp57 protein and activity is required for hemostasis, thrombosis, and platelet accumulation into a growing thrombus (see Abstract and Discussion page 3645). ERp57 is also known as PDIA3.
Nigro teaches that CyPA has a critical function in several human diseases, e.g., cancer cardiovascular etc. (see Abstract and Facts). CyPA is encoded by the peptidyl prolyl isomerase A (PPIA) gene (see page 3 col.2 last paragraph). For instance, CyPA is a critical mediator for cardiovascular diseases and is generally overexpressed in cancer and regulates malignant transformation and metastasis (see Facts). Moreover, CyPA deficient mice are protected from vascular remodeling, atherosclerosis, and thrombosis that complicate the disease (see pages 2-3 Atherosclerosis). Serum CyPA concentrations in unstable angina and acute myocardial infarction subjects are significantly higher than those in patients with stable angina and controls (see page 3 col.2 Cardiac diseases). Myocardial infarction is also known as one of the thrombotic events, which is defined in paragraph 45 in the instant specification. Nigro suggests that an increase in CyPA expression contributes to pathological conditions in light of what is known about CyPA (see page 8 column 1 paragraph 1-2). Therefore, increasing levels of CyPA are shown in both cancers and thrombotic events.
Brown discloses that biomarkers can be assessed for diagnostic, prognostic methods to identify phenotypes, such as a condition or disease, or the stage or progression of a disease, and to determine treatment efficacy (see Abstract). The marker can be from a bodily fluid (e.g., serum or plasma etc.) including nucleic acids, protein, and circulating structures such as vesicles, and nucleic acid-protein complexes (see Abstract, see par.29). The phenotype can also comprise a cardiovascular disease, e.g., pancreatic cancer or a thrombotic event (see par.37 and par.97).
Brown further teaches that the method of identifying a biosignature comprises determining the presence or level of one or more biomarkers in a biological sample. One or more biomarkers are selected from Table 5 which comprises PPIA and PDIA3. Moreover, Brown provides illustrative antigens for use in characterizing various diseases and disorders, e.g., Hsp70 protein for detecting cancers (see par.233 Table 4).
The methods further comprise comparing the biosignature to a reference biosignature, wherein the comparison is used to characterize a cancer. The reference biosignature can be from a subject without cancer (see par.12).
Brown also suggests the methods can be used to assess various diseases or conditions, where biomarkers correspond to various such diseases or conditions. For example, the method can comprise any number of useful biomarkers, e.g., a biosignature can comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more different biomarkers (or in some cases different molecules of the same biomarkers, such protein and nucleic acid) (see par.347). Assessing a plurality of components or characteristics provides increased sensitivity and/or specificity as compared to assessing fewer components or characteristics (see par.330).
Mosawy teaches that platelets play an important role in the development of cardiovascular disease and are essential in the formation of arterial thrombosis. Therefore, antiplatelet is the mainstay treatment for patients who are at high risk of developing thrombotic episodes. However, increasing evidence has indicated inadequate protection by antiplatelet therapy against thrombotic events in some patients. Several studies have therefore investigated many polyphenolic compounds for their cardiovascular and antiplatelet potentials. Quercetin is a polyphenolic flavonol that is found to have potent antiplatelet activity. Quercetin and some of its metabolites Rutin have been demonstrated to inhibit platelet aggregation and activation. (See page 3 col.1 par.2 and page 5 col.2 par.2)
Mosawy states that while quercetin has been demonstrated to reduce or inhibit platelet aggregation, the degree of inhibition and the effective quercetin concentrations are varying. Some studies have shown quercetin to be very effective in reducing platelet aggregation at low concentrations, while other studies have reported higher concentrations were required to achieve anti-aggregatory affects. (See page 3 col.1 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 method of Thalin, by measuring the levels of proteins comprising PPIA, PDIA3 and at least one of the claimed markers, e.g., HSP70 for determining the risk of thrombotic event in a cancer patient because Wang and Nigro teach that ERp57 (or PDIA3) and CypA (or PPIA), respectively, are associated with thrombotic event, wherein the increased expression or levels of these markers contributes to pathological conditions of thrombotic events. Moreover, Brown teaches that PPIA, PDIA3, and HSP70 are the markers for diagnostic, prognostic methods to identify phenotypes, such as a condition or disease, or the stage or progression of a disease, and to determine treatment efficacy, wherein the condition or disease comprises thrombotic events and cancers.
A person of ordinary skill in the art would have been motivated to assess a plurality of biomarkers (e.g., a combination of PPIA, PDIA3, HSP70) because it provides increased sensitivity and/or specificity as compared to assessing fewer components or characteristics (see Brown par.330).
A skilled artisan would have had a reasonable expectation of success in combining Thalin, Wang, Nigro and Brown because Thalin is directed to a method for diagnosing cancer-associated thrombosis, while Brown teaches that determining the presence or level of one or more biomarkers corresponding to various such diseases or conditions in a biological sample can be used to assess various diseases or conditions. Since Wang, Nigro and Brown teach that these biomarkers (e.g., PPIA, PDIA3, HSP70) correspond to cancer and thrombotic events as discussed above, the combined teaching would lead to success in diagnosing cancer-associated thrombosis.
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 treating method of Thalin, by administering to the patient at-risk of thrombotic event quercetin and its derivative compounds as an alternative antiplatelet agent in the method of Thalin (see Thalin page 43 lines 5-11, 22-31, and page 44 lines 1-18). It is because quercetin and its derivative compounds have been demonstrated to reduce or inhibit platelet aggregation as taught by Mosawy. As such, an effective amount of quercetin should be used to achieve anti-aggregatory affects in patients (see Mosawy page 3 col.1 par.2). A person of ordinary skill in the art would have been motivated to use quercetin and its derivative compounds because a conventional antiplatelet therapy against thrombotic events in some patients does not have adequate protection as taught by Mosawy.
Claims 2-3 and 21-22 are rejected under 35 U.S.C. 103 as being unpatentable over Thalin in view of Wang, Nigro, Brown and Mosawy, as applied in claim 1, further in view of Lines et al. (US20130095095).
Claim 2 comprises the limitation in claim 1, detecting the claimed protein in a plasma or serum sample of a patient, and treating the at-risk patient with an effective amount of isoquercetin and optionally an antithrombotic agent.
Claim 3 comprises the limitation in claim 2, and monitoring risk of a thrombotic event in cancer patients weekly bi-weekly, monthly, or as long as indicated throughout the course of treatment.
For claims 2-3, Thalin, Wang, Nigro, Brown and Mosawy teach the invention as discussed in claim 1 above.
The method of Thalin further comprises measuring the presence and/or quantity of a biomarker in plasma samples over a period of time that is indicative of adverse effects in individuals during treatment (see page 4 line 22 and page 26). The period of time is one week or more, or one month or more (see page 26, lines 23-33).
Thalin further teaches that selecting the treatment for the individual is based on presence and/or quantity in the test sample of said biomarkers. The treatment can be thrombosis treatment. See pages 43-44 for lists of antithrombotic agents, e.g., heparin or aspirin.
Wang teaches measuring the level of PDIA3 biomarker in a plasma samples (see page 3643 Flow cytometry and aggregation studies of human platelets).
Nigro teaches measuring the level of PPIA in serum (see page 3 col.2 Cardiac diseases).
Brown teaches measuring the level of biomarkers (e.g., PPIA, PDIA3, or HSP70) in a bodily fluid comprising sera, or plasma (see par.29).
Mosawy teaches using quercetin or rutin as an alternative antiplatelet agent to treat patients with thrombotic event. See discussion in claim 1 above.
See discussion of Thalin, Wang, Nigro, Brown, and Mosawy in claim 1 above.
They do not use an effective amount of isoquercetin and optionally an antithrombotic agent.
Lines teaches a method for improving the efficacy of a blood thinning medication, in which an effective amount of the above-mentioned composition is administered to a subject taking a blood thinning medication. See paragraph 8. The blood thinning medication is an antiplatelet drug or an anticoagulant, e.g., aspirin or heparin (see par.19), which is also called an antithrombotic agent as taught by Thalin.
The method of Lines is for treatment of thrombotic disorders, using a composition containing quercetin, together with one or more of vitamin B3, vitamin C, and folic acid. See Abstract. The thrombotic disorders refer to thrombosis induced by cancer, e.g. the pancreas (see par.20). The quercetin here refers to both quercetin aglycon and quercetin derivatives, e.g., quercetin-3-O-glucoside (isoquercetin) (see par.23). The efficient dose of quercetin is about 250 mg to 1 g (see par.22).
Lines teaches that the efficacy of quercetin is enhanced by vitamin B3, vitamin C, or both because they maintain quercetin levels in plasma up to five times those of quercetin alone (see par.21).
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 method of Thalin, Wang, Nigro, Brown and Mosawy in detecting the claimed proteins in plasma or serum sample with a reasonable expectation of success because Wang, Nigro, and Brown demonstrate the elevated levels of PPIA, PDIA3, or HSP70 in plasma or serum sample of patients with thrombotic event associated with cancer.
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 method of Thalin, treating thrombosis caused by cancer by using a combination of isoquercetin and antithrombotic agent because Mosawy teaches that a conventional antiplatelet therapy against thrombotic events in some patients does not have adequate protection and Lines teaches that the combination of isoquercetin and antithrombotic agent improves the efficacy of antithrombotic agent (Lines par.8).
A skilled artisan would have had a reasonable expectation of success in combining Thalin and Lines because Thalin is directed to a method for diagnosing and treating cancer-associated thrombosis, while Lines refers to a method of improving the efficacy of cancer-related thrombotic disorder treatment.
For claim 21, Thalin, Wang, Nigro, Brown, Mosawy and Lines teach the invention as discussed in claim 2 above, wherein the method is repeated weekly, bi-weekly, monthly, or as long as indicated throughout the course of treatment. The method of Thalin further comprises measuring the presence and/or quantity of biomarker in plasma samples over a period of time is indicative of adverse effects in individuals during treatment (see page 4 line 22 and page 26). The period of time is one week or more, or one month or more (see page 26, lines 23-33).
For claim 22, Thalin, Wang, Nigro, Brown, Mosawy and Lines teach the invention as discussed in claim 2 above, wherein the cancer is lung cancer, pancreatic cancer, colorectal cancer (see Thalin page 38 lines 15-31, Nigro page 5 col.2 par.2).
Claims 5-6 are rejected under 35 U.S.C. 103 as being unpatentable over Thalin in view of Wang, Nigro, Brown, Mosawy and Lines, as applied in claim 2, and further in view of Rohatgi et al. (Vascular Toxicity Associated With Chemotherapy and Molecular Targeted Therapy: What Should a Radiologist Know?, Vascular and Interventional Radiology, vol.203, issue 6, 2014).
For claims 5-6, Thalin, Wang, Nigro, Brown, Mosawy and Lines teach the invention as discussed in claim 2 above. The method of Thalin is used for screening for cancer-associated thrombosis or an elevated risk for cancer-associated thrombosis in cancer individuals, wherein the results indicate cancer-associated thrombosis or an elevated risk for cancer-associated thrombosis in said cancer individuals (see page 5 lines 15-33, page 6 lines 6-11).
Thalin further discloses that chemotherapy is known to increase the risk for thrombosis. Also, the use of anticoagulant therapy, e.g., heparin, is associated with a substantial risk of bleeding, especially in patients with active cancer. So the method of predicting the susceptibility to cancer-associated thrombosis in cancer patients could reveal patients with the highest risk of thrombosis and allow for individualized thrombo-prophylaxis strategies. See page 51 paragraph 1.
However, Thalin does not explicitly teach the method of predicting the susceptibility to cancer-associated thrombosis in cancer patients is done on the patient exhibiting no primary venous thromboembolism (as recited in claim 5), or no major hemorrhages during treatment (as recited in claim 6).
Wang teaches that platelet surface ERp57 (or PDIA3) targeting aIIbb3 is required for platelet incorporation into a developing thrombus (see page 3643 col.1 par.1).
Nigro teaches that CyPA (or PPIA) is involved in the early phase of atherosclerosis (see page 3 col.1 par.1 col.2 par.1) and the elevated level of PPIA is observed in acute myocardial infarction subjects (page 3 col.2 par.5).
Brown also teaches that the method of detecting one or more biomarkers is used for detecting a disease or condition, including pre-symptomatic early stage detection (see par.88).
Rohatgi clearly points out that venous and arterial thromboembolic events and rarely hemorrhage are complications of chemotherapy (see Abstract). There are some severe adverse events associated with the toxicity of therapeutic agents. For example, grades 3 or 4 hemorrhage or high-grade venous thromboembolism (see page 1354 col.1 last par. col.2 2nd par.). An awareness of modern-day drug toxicities is key to the early diagnosis and management of these complications. Many affected patients are initially asymptomatic, and thrombosis is incidentally discovered on routine imaging examinations performed for re-staging. Early diagnosis improves outcomes because patients benefit from early intervention. See Conclusion page 1356.
Therefore, it would have been obvious to one of ordinary skill in the art before the effective filing date of the claimed invention to apply the method of Thalin to screen on the cancer patients having no primary venous thromboembolism, or no major hemorrhages for the risk of thrombotic events for predicting the susceptibility to cancer-associated thrombosis in cancer patients because Thalin and Rohatgi teach that chemotherapy is known to increase the risk for thrombosis, wherein Rohatgi teaches that venous and arterial thromboembolic events and rarely hemorrhage are complications of chemotherapy.
A person of ordinary skill in the art would have been motivated to screen on the cancer patients showing no clinical symptoms of severe thrombosis (e.g., primary venous thromboembolism, or major hemorrhages) because early diagnosis will lead to early intervention, thereby improving a patient outcome (see Thalin page 51 paragraph 1, Rohatgi Conclusion page 1356).
A skilled artisan would have had a reasonable expectation of success in detecting the claimed proteins in the cancer patients showing no clinical symptoms of severe thrombosis because these proteins are involved in the early phase of thrombotic events or cancers, as taught by Wang, Nigro and Brown.
Claims 23-25 are rejected under 35 U.S.C. 103 as being unpatentable over Thalin in view of Wang, Nigro, Brown, Mosawy and Lines, as applied in claim 21, and further in view of Karimi et al. (Cancer-Associated Thrombosis, The Open Cardiovascular Medicine Journal, 2010, 4, 78-82).
For claims 23-25, Thalin, Wang, Nigro, Brown, Mosawy and Lines teach the invention as discussed in claim 21 above. They do not teach the cancer is a metastasizing cancer.
Karimi teaches that thrombosis is a common complication in patients with cancer (see Abstract), wherein the cancer is pancreas cancer, or breast cancer, or colorectal cancer (see page 78 col.1 par.3, and page 80 col.1 par.3). It is estimated that about 4-20% of patients with cancer experience venous thrombosis (page 80 col.1 par.2). The appearance of venous thrombosis has been clearly associated with metastatic cancer and the stage of cancer (see page 80 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 apply the modified method of Thalin to cancer patients, wherein the cancer is colorectal cancer, pancreatic cancer, or breast cancer, or wherein the cancer is a metastasizing cancer from colorectal cancer, pancreatic cancer, or breast cancer. It is because Karimi teaches that thrombosis is highly associated with these types of cancer and with metastatic cancers as discussed above.
Claim(s) 14-20 is/are rejected under 35 U.S.C. 103 as being unpatentable over Thalin (WO2017027379) in view of Wang et al. (Platelet-derived ERp57 mediates platelet incorporation into a growing thrombus by regulation of the αIIbβ3 integrin, Blood (2013) 122 (22): 3642–3650), Nigro et al. (Cyclophilin A: a key player for human disease, Cell Death and Disease (2013) 4, e888) Brown (US20140220580), Kim (US20100203510), Beretta (US20070202496), and Lu et al. (Identification and quantification of Heat-Shock Protein 70, BioProcess International 13(9) Oct 2015).
For claims 14, 16 and 18, Thalin, Wang, Nigro, and Brown teach a method for detecting the protein markers PPIA, PDIA3 and at least one of EIF5A, EIF4H, EIF4a3, UBE2N, UBE2L3, UBE2I, and HSP70 in a plasma sample from a subject with cancer, wherein the subject is actively undergoing treatment for cancer, the treatment comprising chemotherapeutic treatment and/or radiation, and/or immunotherapy, and/or cell therapy (see discussion of Thalin, Wang, Nigro, and Brown in claim 1 above).
Thalin teaches an ELISA-based assay to detect a target biomarker comprising: using a binding agent capable of binding to the biomarker (e.g., antibody) (see page 31, lines 17-33), a solid support coated with the binding agent capable of binding to the biomarker (page 32 lines 20-25), antibody conjugates (e.g., anti-rabbit horseradish peroxidase) are reactive with the antibodies specific to the biomarker (page 33 lines1-5). The method comprises contacting the sample with the solid support and the conjugates and assaying a reaction of the conjugates with the sample (see page 32 lines 20-33, page 33 lines 1-15).
Thalin does not specifically teach the methods of detection are for detecting PPIA, PDIA3, and HSP70.
Kim discloses a method of detecting and quantifying PPIA in a serum sample comprising: anti-PPIA antibodies, and antibody conjugated HRPs that are reactive with the antibody to PPIA (see par.41, 66 and 70-71). See par.70: after coating the polyclonal antibody obtained from a rabbit with 1 μg/ml of a sample, the coated antibody was blocking treated using 1% BSAT. See par.71: after adding a PPIA sample solution followed by a monoclonal antibody conjugated HRP to the treated antibody, the antibody was developed using TMB (3.3′, 5.5′-tetramethylbenzidine).
Beretta discloses a method of detecting and quantifying PDIA3 by Elisa comprising: anti-ERp57 antibody is coated on the plate, adding serum sample, detecting with anti-Erp57 SPA-725 antibodies, and antibody conjugate (see par.19, par.46: teaching that ninety six-well microplates are coated with anti-ERp57 N-20 antibodies, antibody conjugate is horseradish peroxidase-conjugated goat anti-mouse immunoglobulin which is reactive with anti-Erp57 SPA-725 antibodies).
Lu discloses a method of detecting and quantifying HSP70 comprising: a solid support coated with antibodies to Hsp70 (see page 50 col.3 par.4 teaching anti-Hsp70 coated wells), antibody conjugates that are reactive with the antibodies to Hsp70 (see page 50 col.3 par.4 teaching that HRP-conjugated goat-anti rabbit IgG is reactive with a polyclonal anti-Hsp70 detection antibody).
Lu discloses a method of detecting and quantifying HSP70 comprising: a solid support coated with antibodies to Hsp70 (see page 50 col.3 par.4 teaching anti-Hsp70 coated wells), antibody conjugates that are reactive with the antibodies to Hsp70 (see page 50 col.3 par.4 teaching that HRP-conjugated goat-anti rabbit IgG is reactive with a polyclonal anti-Hsp70 detection antibody).
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 method of Thalin, substituting the antibodies to the target antigen H3Cit with the antibodies specific to PPIA, PDIA3, and HSP70 for measuring the biomarkers PPIA, PDIA3, and HSP70 in plasma or serum from a patient with cancer. The motivation to detect these biomarkers in the sample is for the purpose of early assessing various diseases or conditions, e.g., thrombotic events in cancer patients. It will help to improve patient outcomes due to an early intervention (see Thalin page 51 paragraph 1 teaching that quantification of biomarkers in plasma could reveal patients with the highest risk of thrombosis and allow for individualized thrombo-prophylaxis strategies, see discussion of Thalin, Wang, Nigro, and Brown in claims).
A skilled artisan would have had a reasonable expectation of success in detecting PPIA, PDIA3, and HSP70 by using the method of Thalin because Kim, Beretta, and Lu support the detection of PPIA, PDIA3, and HSP70 by providing the detection reagents (e.g., anti-PPIA antibody, anti-PDIA3 antibody, anti-HSP70 antibody, antibody conjugates that are reactive with the antibodies to the target proteins) and the immunoassay of detecting target antigen comprising a solid support coated with the antibody to the target antigen, and antibody conjugates that are reactive with the antibody to the target antigen.
For claim 15, Thalin, Wang, Nigro, Brown, Kim, Beretta, and Lu teach the method of claim 14. Thalin teaches the conjugate comprises HRP or alkaline phosphatase (see page 45 lines 21-23). Thalin teaches that TMB is a chromogenic substrate of HRP enzyme (see page 33 line 9).
Thalin does not teach using antigen standard comprising PPIA, PDIA3, and HSP70.
Lu discloses a method of detecting and quantifying HSP70 further comprising Hsp70 standard (see page 50 col.3 par.4), wherein the standard antigen is used for quantifying the concentration of the target antigen in the sample (see page 56 col.1 par.2-3).
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 method of Thalin, further comprising an antigen standard of the target protein markers PPIA, PDIA3, and HSP70 for the benefit of quantifying the concentration of the target protein markers with a reasonable expectation of success because Lu supports the method of using the antigen standard as a reference signal to calculate the concentration of the target antigen (see page 56 col.1 par.2-3). One having an ordinary skill in the art would have been motivated to quantify the concentration of the target protein markers for the benefit of defining the elevated levels of the protein markers among cancer subjects to determine the risk for thrombotic events in the subjects.
For claim 17, Thalin, Wang, Nigro, Brown, Kim, Beretta, and Lu teach the method of claim 14. Thalin teaches that the solid support is microplate (see page 34 lines 20-25). Thalin does not teach that the solid support is a microbead array.
Brown teaches an immunoassay to detect biomarkers, comprising contacting the sample with a solid support coated with an antibody to a marker and the conjugates and assaying the reaction of the conjugates with the sample (see par.33, 177-179: teaching that micro-vesicle may be contacted with one or more binding agent, see par.177-179: vesicle expressing biomarkers is detect with a binding agent linked directly or indirectly to a solid surface or substrate, wherein the binding agent is an antibody, see par.599 and FIG. 2A showing an antibody, the detector antibody, is directly coupled to a label, and is used to detect a biomarker on the captured vesicle). The substrate can be microwell plate (see par.427), microbead array (see par.436). The chromogenic label can comprise alkaline phosphatase with para-nitrophenyl phosphate (see par.186). The method used to detect biomarkers comprises immunoassay, ELISA (see par.32).
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 method of Thalin, substituting the solid support of Thalin with the microbead array as taught by Brown with a reasonable expectation of success because the microplate is functionally equivalent to the microbead array in providing a solid support to immobilize a capture antibody.
For claim 19, Thalin, Wang, Nigro, Brown, Kim, Beretta, and Lu teach the method of claim 14. Thalin teaches the conjugate comprises HRP or alkaline phosphatase (see page 45 lines 21-23). Thalin does not teach a chromogenic reagent for alkaline phosphatase comprising p-nitrophenyl phosphate.
Brown teaches that the chromogenic label can comprise alkaline phosphatase with para-nitrophenyl phosphate (see par.186).
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 method of Thalin, substituting enzyme HRP with enzyme alkaline phosphatase conjugate with a reasonable expectation of success because the alkaline phosphatase label is functionally equivalent to the HRP in providing a color signal for the detection method. Accordingly, the chromogenic label used for alkaline phosphatase is para-nitrophenyl phosphate as taught by Brown.
For claim 20, Thalin, Wang, Nigro, Brown, Kim, Beretta, and Lu teach the method of claim 19. Claim 20 recites contacting the antigen standards with the solid support and the conjugates, and assaying the relative levels of PPIA, PDIA3 and at least one of EIF5A, EIF4H, EIF4a3, UBE2N, UBE2L3, UBE2I, and HSP70 in the sample relative to the antigen standards. It generally means that the antigen standard is for generating the standard curve of the antigen concentration so that the amount of a tested antigen detected from a test sample is calculated in relative to the antigen standard curve.
Lu discloses a method of detecting and quantifying HSP70 further comprising Hsp70 standard (see page 50 col.3 par.4), wherein the standard antigen is used for quantifying the concentration of the target antigen in the sample (see page 56 col.1 par.2-3). The chemiluminescence signal from the antibody conjugates can be quantified and compared with a signal generated from a series of standards (i.e., standard curve) (see page 56 col.1 par.2). The sample is run at appropriate dilutions and graphed its chemiluminescence signals with a linear-fitted standard curve, then interpolated the HSP70 concentration from the sample (see page 56 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 method of Thalin, further comprising an antigen standard of the target protein markers PPIA, PDIA3, and HSP70 and assaying relative levels of PPIA, PDIA3, and HSP70 in the sample relative to the antigen standards for the benefit of quantifying the concentration of the target protein markers with a reasonable expectation of success because Lu supports the method of using the antigen standard as a reference signal to calculate the concentration of the target antigen (see page 56 col.1 par.2-3). One having an ordinary skill in the art would have been motivated to quantify the concentration of the target protein markers for the benefit of defining the elevated levels of the protein markers among cancer subjects to determine the risk for thrombotic events in the subjects.
Response to Arguments
For rejection under 35USC 112(a), Applicant's arguments filed 12/18/2025 have been fully considered and they are persuasive because the claimed methods require detecting "elevated levels of PPIA, PDIA3 and at least one ofEIF5A, EIF4H, EIF4a3, UBE2N, UBE2L3, UBE2I, and HSP70" as recited by claims 1-3; PDIA3 is only recited in combination with other biomarkers. Even if PDIA3 expression varies across cancer types, the panel includes multiple biomarkers, and "diagnosing the patient as at-risk of a thrombotic condition when PPIA, PDIA3 and at least one of EIF5A, EIF4H, EIF4a3, UBE2N, UBE2L3, UBE2I, and HSP70 are elevated" as recited by claims 1-3 would still be enabled through the combination of biomarkers in the panel.
For rejection under 35USC 103, Applicant’s arguments with respect to claim(s) 1-2, 5-6, 14-20 have been considered but are moot because the new ground of rejection is made in view of the amendment of the claims.
Conclusion
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.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to CHAU N.B. TRAN whose telephone number is (571)272-3663. The examiner can normally be reached Mon-Fri 8:30-6:30 CT.
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, Bao-Thuy L Nguyen can be reached at 571-272-0824. The fax phone number for the organization where this application or proceeding is assigned is 571-273-8300.
Information regarding the status of published or unpublished applications may be obtained from Patent Center. Unpublished application information in Patent Center is available to registered users. To file and manage patent submissions in Patent Center, visit: https://patentcenter.uspto.gov. Visit https://www.uspto.gov/patents/apply/patent-center for more information about Patent Center and https://www.uspto.gov/patents/docx for information about filing in DOCX format. For additional questions, contact the Electronic Business Center (EBC) at 866-217-9197 (toll-free). If you would like assistance from a USPTO Customer Service Representative, call 800-786-9199 (IN USA OR CANADA) or 571-272-1000.
/CHAU N.B. TRAN/Examiner, Art Unit 1677
/BAO-THUY L NGUYEN/Supervisory Patent Examiner, Art Unit 1677
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