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 1-31 are currently pending.
Election/Restrictions
2. Applicant’s election without traverse of Group I, claims 1-14 and 22-31 in the reply filed on November 14, 2025 is acknowledged.
The Office would like to note that it appears the previous Examiner inaccurately accounted for the pending claims. Specifically, as it pertains to claims 22-31. Group I (claims 1-14 and 22-31) is identified as being drawn to method for detecting anti-Bartonella henselae antibodies. Group II (claims 15-21) is identified as being drawn to kits comprising Bartonella henselae antigens. However, claims 22-31 are not drawn to a method for detecting anti-Bartonella henselae antibodies. In fact claims 22-31 are drawn to a method of treating a human subject for Bartonella henselae infection. Claims 22-31 are drawn to a completely different invention from what is encompassed by Group I. The methods as evidenced by the claims themselves, are directed to different inventions which are not connected in design, operation, or effect. These methods are independent since they are not disclosed as capable of use together, they have different modes of operation, they have different functions, and they have different effects.
For these reasons, the Office believes claims 22-31 were erroneously placed in Group I and thus should not be examined with the claims drawn to methods for detecting. Consequently, claims 15-31 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected invention, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on November 14, 2025.
Claims 1-14 are currently under examination.
Information Disclosure Statement
3. The information disclosure statement (IDS) submitted on June 29, 2023 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.
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.
4. Claim(s) 1-5 and 7-14 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Huang et al., WO 2010/021849 A2; Published: 2/25/10.
Independent claim 1 is drawn to a method of preparing a calibration sample for detection of anti-Bartonella henselae antibodies in a subject, comprising obtaining one or more control sample from one or more human control subject infected with Bartonella henselae, isolating antibodies from the one or more control sample to obtain a solution comprising isolated antibodies, affinity purifying, from the isolated antibodies, antibodies that specifically bind target Bartonella henselae 17 kDa antigen or a fragment thereof to obtain a solution of purified antibodies, and diluting the solution of purified antibodies to a predetermined protein concentration to obtain the calibration sample.
Independent claim 7 is drawn to a method of detecting anti-Bartonella henselae antibodies in a test subject, comprising contacting a peptide with a subject sample, comprising antibodies, from the test subject, wherein the peptide comprises a test Bartonella henselae 17 kDa antigen or a fragment thereof, and detecting a test signal indicating subject sample antibodies binding to the peptide; contacting the peptide with a calibration sample, comprising antibodies, wherein the calibration sample comprises a threshold protein concentration, and detecting a calibration signal indicating calibration sample antibodies binding to the peptide, wherein the calibration sample was prepared by: obtaining one or more control sample from one or more human control subject infected with Bartonella henselae, isolating antibodies from the one or more control sample to obtain a solution comprising isolated antibodies, affinity purifying, from the isolated antibodies, antibodies that specifically bind a target Bartonella henselae 17 kDa antigen or a fragment thereof to obtain a solution of purified antibodies, diluting the solution of purified antibodies to a predetermined protein concentration to obtain a base calibration sample, and diluting the base calibration sample to obtain the calibration sample having a threshold protein concentration; wherein detecting anti-Bartonella henselae antibodies in the test subject comprises detecting a test signal that equals or exceeds the calibration signal.
Huang et al. disclose methods for the detection of antibodies specific for Bartonella henselae. The 17-kDa polypeptides, and methods are useful in the detection of recent and/or ongoing infections with Bartonella henselae (see abstract; meets claim 1, 7 and 14). The method for the detection of antibodies specific for Bartonella henselae in a biological sample (i.e. serum ) from a mammal, preferably a human (see paragraph 0075). The method comprises the steps of binding a recombinant polypeptide according to the disclosure to a solid support; contacting the bound recombinant polypeptides to said sample to form antibody-antigen complexes if said antibodies are present in said sample; washing away unbound immunoglobulin; and detecting the presence of any formed antigen-antibody complexes, such as by contacting such complexes with a secondary antibody capable of initiating a visualization reaction, such as a color change of a substrate (see paragraph 0025-27; meets claim 7).
Steps were taken to solubilize the protein. Purification of said protein was conducted under denaturing conditions. Nickel chelating affinity chromatography was used to capture the protein via interaction with the 6x his tag. Briefly, recombinant protein was released from the cell using BugBuster MasterMix. The cell pellet from spin down was re-suspended in 5ml BugBuster MasterMix by gentle pipette mixing. The mixtures were centrifuged to separate insoluble part from soluble component. Both soluble and insoluble fraction was transferred to a fresh tube for SDS PAGE analysis (see paragraph 00121; meets claim 1 and 7).
ROC accesses is the performance of the system in terms of "Sensitivity" and "1 -Specificity" for each observed value of the discriminator variable assumed as decision threshold (i.e., cutoff value to differentiate between two groups of response). For ELISA, the cutoff value can be shifted over a range of observed values (i.e., OD450nm reading), and Sensitivity and 1- Specificity can be established for each of these values (meeting claim 1 and 7). Moreover, Huang discloses that the detection of antibody binding in IFA sero-positive sera may be accomplished by techniques known in the art, e.g., ELISA (enzyme-linked immunosorbent assay), western blots, and the like. In one embodiment, antibody binding is assessed by detecting a label on the primary antibody. In another embodiment, the primary antibody is assessed by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labeled. Many means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention (see paragraph 0092; meets claims 1, 7 and 13). The present invention further comprises a positive and negative serum control (see paragraph 0093). A significant difference between IFA sero-positive sera and IFA-negative sera at coating concentration from 100 ng/ml to 16 ng/ml and a sera dilution of 1:100. The capacity of the recombinant protein to differentiate positive sera to negative sera was enhanced at higher coating concentrations from 12 ng/ml to 500 ng/ml. From this analysis, it was determined that 250 ng/ml coating antigen (recombinant Fragment 3) was sensitive enough to specifically identify antibodies (see paragraph 00136; meets claim 12).
Further, in this series of studies, a panel containing 12 IFA positive serum samples and 6 IFA negative serum samples was evaluated by ELISA using the respective antigen as the coating antigen. Figure 15 presents the results of this ELISA analysis (see paragraph 00139; meets claim 1, 5, 7 and 11). An IgG ELISA assay was adopted and evaluated the binding activity of the modified peptides towards IgG. The ELISA procedure involves: (i) coating 96-well micro- titer plates with modified peptides at varying concentrations (e.g., 0.2-5 μg/ml) at 40C overnight; (ii) adding 5% skim milk to block non-specific binding; (iii) adding patients' sera to allow formation of antibody-antigen complex; (iv) detecting the antibody-antigen complex. IFA sero-positive sera served as positive controls and IFA sero-negative sera served as negative controls. Detection of antibody-antigen complex was performed (00159; meets claim 1 and 7).
In an individual Patient Study a peptide exhibited a dose-dependent increase in binding towards sero-positive serum as measured by OD45onm- These data indicate that the amino acid sequence of peptide 3C is responsible for the specific recognition of antibodies that is present in IFA-sero positive sera. (see paragraph 00160). A panel of 47 human sera was used in an ELISA assay to evaluate the sensitivity and specificity for synthetic polypeptides of 17-kDa protein. This panel contains 27 IFA positive sera and 20 IFA negative sera (see paragraph 00165; meets claim 5, 11, 13).
Further, for the IgM Capture assay, purified recombinant 17-kDa full-length protein was biotinylated using the Sulfo-NHS Biotin kit. MaxiSorp 96 well plates were coated with anti-human IgM at a concentration of 1 μg/ml in coating buffer (0.015 M Na2CO3, 0.035 M NaHCO3 [pH 9.6]) and incubated overnight at 4°C. The plates were washed with PBS-Tween 20 and blocked with 1% bovine serum albumin (BSA) at room temperature. IgM positive and negative patient sera were used. The sera were diluted 1 : 100 in 1% BSA with 0.05% Tween 20, prior to being run in duplicate, and reacted with the plates for 1 hr at room temperature. Biotinylated recombinant 17-kDa protein was added to the plates at 1 μg/ml in dilution buffer and incubated at room temperature for 1 h with shaking. Streptavidin-HRP was used to detect the plates followed by development with 3,3',5,5'-Tetramethylbenzidine (TMB) (Moss, Pasadena, MD) for 30 minutes. The reaction was stopped with 2M HCl and the absorbance at 450 nm was recorded after a period of 5-30 minutes (see paragraph 00167; meets claims 1 and 7).
The results showed that recombinant 17-kDa full-length protein of Bartonella henselae could bind to IgM IFA-positive (n=13) and IFA-negative (n=34) patient samples with sensitivity and specificity values of 100% and 97.1%, respectively. Receiver operating characteristic (ROC) curve analysis (determines threshold and cutoff values) was used to evaluate the performance of the IgM Capture method. ROC plots can be used to compare all tests, even when tests have quite different cutoff values. The study of area under the curve (AUC), which evaluates the probabilities of (1 -Specificity) and Sensitivity, is an important statistical feature of such curves. ROC analysis of the calibrated slopes revealed an AUC of 0.998 (95% CI, 0.919-1.000) (see paragraph 00168; meets claims 5, 7 and 11). During the IgG ELISA Analysis, the reactions were visualized by adding 100 μl substrate 3,3'5,5'-Tetramethylbenzidine Solution (see paragraph 00177; meets claim 7).
Lastly, Huang et al. disclose that the target Bartonella henselae 17 kDa antigen comprises a sequence of amino acids and the sequence is 100% identical to the claimed SEQ ID NOs: 1-13 (see attached SCV result; meets claims 2-4 and 8-10).
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SEQ ID NO: 1
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SEQ ID NO: 2
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SEQ ID NO: 3
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SEQ ID NO: 4
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SEQ ID NO: 5
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SEQ ID NO: 6
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SEQ ID NO: 7
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SEQ ID NO: 8
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SEQ ID NO: 9
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SEQ ID NO: 10
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109
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SEQ ID NO: 11
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114
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SEQ ID NO: 12
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113
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SEQ ID NO: 13
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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.
5. Claim(s) 6 is rejected under 35 U.S.C. 103 as being unpatentable over Huang et al., WO 2010/021849 A2; Published: 2/25/10 as applied to claims 1-5 and 7-14 above, and further in view of Smith, FFPE or Frozen? Working with Human Clinical Samples; 2014; Life Science Articles.
Independent claim 1 is drawn to a method of preparing a calibration sample for detection of anti-Bartonella henselae antibodies in a subject, comprising obtaining one or more control sample from one or more human control subject infected with Bartonella henselae, isolating antibodies from the one or more control sample to obtain a solution comprising isolated antibodies, affinity purifying, from the isolated antibodies, antibodies that specifically bind target Bartonella henselae 17 kDa antigen or a fragment thereof to obtain a solution of purified antibodies, and diluting the solution of purified antibodies to a predetermined protein concentration to obtain the calibration sample.
Independent claim 7 is drawn to a method of detecting anti-Bartonella henselae antibodies in a test subject, comprising contacting a peptide with a subject sample, comprising antibodies, from the test subject, wherein the peptide comprises a test Bartonella henselae 17 kDa antigen or a fragment thereof, and detecting a test signal indicating subject sample antibodies binding to the peptide; contacting the peptide with a calibration sample, comprising antibodies, wherein the calibration sample comprises a threshold protein concentration, and detecting a calibration signal indicating calibration sample antibodies binding to the peptide, wherein the calibration sample was prepared by: obtaining one or more control sample from one or more human control subject infected with Bartonella henselae, isolating antibodies from the one or more control sample to obtain a solution comprising isolated antibodies, affinity purifying, from the isolated antibodies, antibodies that specifically bind a target Bartonella henselae 17 kDa antigen or a fragment thereof to obtain a solution of purified antibodies, diluting the solution of purified antibodies to a predetermined protein concentration to obtain a base calibration sample, and diluting the base calibration sample to obtain the calibration sample having a threshold protein concentration; wherein detecting anti-Bartonella henselae antibodies in the test subject comprises detecting a test signal that equals or exceeds the calibration signal.
Huang et al. teach the limitations as it pertains to methods for the detection of antibodies specific for Bartonella henselae, as set forth supra.
Huang et al. do not specifically teach the furthering step limitation, where freezing the calibration sample takes place, as recited in claim 6.
Smith teaches that frozen samples can preserve specimens well depending upon your application. Specifically, freezing has the advantage that it is fast, compared to FFPE sample preparation, and the resulting samples are well suited for molecular analysis. For molecular analysis, including work with DNA, RNA and post-translational protein modifications (PTMs), frozen samples are preferable to FFPE samples for several reasons. The variability that nonstandardized preparation methods introduce into FFPE samples affects the integrity of molecular data disproportionately. For subsequent molecular analyses, such as mass spectrometry, quantitative real-time PCR, next-generation sequencing and Western blotting for PTMs, frozen samples are a necessity. “Frozen sections are considered the gold standard for most molecular assays, especially [for sequencing DNA or RNA] strands that are longer than 50 base pairs,” says Jennifer Freeland, research and development manager at Thermo Fisher Scientific (which recently acquired Life Technologies). Also, Freeland says, “frozen tissue is best when experimenting with new antibodies and immunohistochemistry results are unknown.”
It would have been obvious before the effective filing date of the presently claimed invention to freeze a sample because it offers the advantage in that it is fast resulting in samples well suited for molecular analyses as suggested by Smith with a reasonable expectation of success. This modification may be viewed as an additional element which is known in the prior art. One skilled in the art could have combined the elements as claimed by known methods with no change in their respective functions, and the combination would have yielded predictable results to one of ordinary skill in the art at the time of the invention.
Further, it would be obvious and the combination would have yielded predictable results to one of ordinary skill in the art because freezing biological samples preserves their integrity by halting metabolism, making them ideal for long-term storage, molecular analysis (DNA, RNA, proteins), and future research, offering advantages like improved data quality, reduced contamination risk, efficient processing, and better preservation of morphology and molecular structures compared to other methods. Lastly, this technique is crucial for biobanking, diagnostics, and scientific studies, ensuring samples remain viable and consistent over time.
Accordingly, the subject matter of claim 6 would have been prima facie obvious to one of ordinary skill in the art before the effective filing date of the presently claimed invention, absent evidence to the contrary.
Conclusion
6. No claim is allowed.
7. The prior art made of record and not relied upon is considered pertinent to applicant's disclosure. Eskow et al., JAMA Neurology, 2001; 58(9): 1357-63.
8. 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, Vanessa Ford can be reached at (571) 272-0857. 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 January 8, 2026
/BRIAN GANGLE/Primary Examiner, Art Unit 1645