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 OFFICE ACTION
This Office Action is in response to the papers filed on 24 October 2025.
APPLICANT’S ELECTION
Applicants’ election without traverse of Group I (Claims 1, 2, 19, 22-23, 134-135, 140 and 360-371; drawn to a method of extracting one or more neutrophil serine proteases from a sample comprising white blood cells obtained from a subject) in the reply filed on 24 October 2025 is acknowledged. Claims directed to the non-elected invention have been cancelled.
CLAIMS UNDER EXAMINATION
Claims 1-2, 19, 22-23, 134-135, 140 and 360-371 are pending and have been examined on their merits.
PRIORITY
Provisional Application 63/053939, filed on 20 July 2020, is acknowledged.
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.
Claims 1-2, 19, 22-23, 134-135, 140, 360-361 and 365-371 are rejected under 35 U.S.C. 103 as being unpatentable over Seren et al. (Consequences of cathepsin C inactivation for membrane exposure of proteinase 3, the target antigen in autoimmune vasculitis. J. Biol. Chem. (2018) 293(32) 12415–12428) in view of Kalupov et al. (Structural Characterization of Mouse Neutrophil Serine Proteases and Identification of Their Substrate Specificities. JBC. Vol. 284, No. 49, pp. 34084–34091, 2009).
Seren studies neutrophil lysates from patients (see page 12415, right column, first paragraph). Blood is collected and white blood cells (a sample comprising WBCs) are pelleted (see page 12425, let column, second paragraph). Neutrophils (hence, WBCs) are isolated and purified. Isolated neutrophils are suspended in PBS (phosphate buffered saline) (see page 12415, left column, third paragraph). Therefore Seren adds an aqueous wash solution to form a mixture of the aqueous wash solution and the sample, wherein the wash solution is a phosphate buffered saline.
Neutrophils are lysed in HEPES buffer containing NaCl and 0.05% Nonidet P-40 (see page 12415, right column, second paragraph). As evidenced by the specification, the IUPAC name of Nonidet P-40 is octylphenoxypolyethoxyethanol (see [0047] of PG Pub). Therefore the art teaches adding a first aqueous medium comprising at least 0.01% of octylphenoxypolyethoxyethanol to cells to obtain a first lysate. Because Seren adds the claimed first aqueous medium to white blood cells, it would be expected to produce a first cell lysate comprising a first NSP extract and a first WBC residual.
Seren teaches centrifugation (at 10,000g for 10 min) to separate soluble fractions from cell debris (same cited section). Seren teaches the soluble fraction contains proteins (page 12425, right column, second paragraph). Seren teaches PR3 is soluble (see page 12422, right column, second paragraph). Therefore centrifugation is interpreted to separate the first lysate from a WBC residual (cell debris) to provide a first separated cell lysate.
The deficiencies of Seren are:
Seren forms a mixture containing WBCs and an aqueous wash solution comprising PBS. The art is silent regarding centrifuging the mixture to form a supernatant and a pellet.
Seren lyses white blood cells with the first aqueous medium. The art does not teach repeating the lysis and separation steps to produce a second cell lysate, a second WBC residual and a second NSP extract.
Kalupov teaches a method of purifying neutrophil elastase (NE), cathepsin G (CG), and proteinase 3 (Pr3) (see page 34086, right column, second paragraph).
Neutrophils are isolated from a subject (page 34085, left column, fifth paragraph).
Lavage fluid was collected from a patient and red blood cells were removed (page 34085, left column, fifth paragraph). Viable cells in the lavage fluid are cytospun (centrifuged) (page 34085, left column, fifth paragraph). The cell pellet is resuspended in PBS (same cited section). Therefore Kalupov forms a mixture of white blood cells and aqueous wash solution. To purify neutrophil serine proteases, cell pellets were subjected to three cycles of freezing and thawing. The cell suspension was centrifuged after each cycle, and supernatants were collected and diluted in two volumes of 50 mM phosphate-buffered saline, pH 7.4. Pooled supernatants were run on a heparin-sepharose column to recover protease fractions (page 34085, left column, last paragraph).
The following is also taught by the art:
Kalupov et al. teach Neutrophil serine proteases (NSPs), neutrophil elastase (NE), cathepsin G (CG), and proteinase 3 (Pr3), are mainly stored in neutrophil primary granules in readily active forms. NSPs are structurally related (page 34084, left column, first paragraph).
It would have been obvious to centrifuge the mixture of WBCs and aqueous wash solution taught by Seren. One would have been motivated to do so since Kalupov teaches centrifuging after adding PBS to neutrophils. The skilled artisan would do so to pellet neutrophils for subsequent lysis. One would have been motivated to collect the supernatant since Kalupov teaches collecting the supernatant to isolate NSPs.
One would have had a reasonable expectation of success since Kalupov teaches washed neutrophils can be centrifuged, and NSPs can be isolated from supernatants produced following centrifugation. It would have been obvious to repeat the lysis and separation steps to produce a second cell lysate, a second WBC residual and a second NSP extract. One would have been motivated to do so since Kalupov teaches repeatedly lysing, centrifuging and separating lysates to isolate NSPs. One would have expected similar results since both references are directed to methods of isolating NSPs from neutrophils. Because the claimed method is rendered obvious, the supernatant and NSP extracts would be expected to contain one or more NSPs. Therefore claim 1 is rendered obvious.
Regarding claim 2: Seren teaches lysis at “room temperature” (see text below Figure 1). Because the term “about” is not defined in the claims or the specification, the temperature taught by Seren reads on claim 2.
Repeating the cell lysis steps is rendered obvious on the grounds set forth above. Seren teaches an aqueous medium comprising 0.05% octylphenoxypolyethoxyethanol. It would have been obvious to use the same medium. One would have been motivated to do so since Seren teaches it lyses cells. Therefore claim 19 is included in this rejection.
It would have been obvious to combine the teachings of the prior art by pooling the first and second separated lysates. One would have been motivated to do so since Kalupov teaches pooling to isolate NSPs. One would have had a reasonable expectation of success since Kalupov teaches NSPs can be isolated from pooled fractions. One would have expected similar results since both references are directed to methods of isolating NSPs from neutrophils. Therefore claim 22 is rendered obvious.
Kalupov teaches centrifuging after each cycle, collecting the supernatant (hence, the lysate) and pooling to analyze protease activities (page 34086, left column, first paragraph). Therefore claim 23 is included in this rejection.
Regarding claim 134: The art teaches an aqueous medium comprising 50 mM HEPES buffer containing 750 mM NaCl, and 0.05% Nonidet P-40 (octylphenoxypolyethanol)
(see page 12415, right column, second paragraph). The specification states the term “about” is understood to mean those values near to a recited value ([0081] of PG Pub). Therefore the amounts of Herpes, NaCl and octylphenoxypolyethoxyethanol taught by Seren read on the claim. Claim 134 is included in this rejection.
The method of claim 1 is rendered obvious. Kalupov teaches NE can be isolated (supra). Therefore claim 135 is included in this rejection.
Seren isolated neutrophils from human subjects with Papillon–Lefèvre syndrome (PLS) (see page 12417, left column, first paragraph of Results; see page 1402 “Isolation of neutrophils from human peripheral blood”). Therefore claim 140 is included in this rejection.
Kalupov analyzes active NSP in each supernatant (see page 34086, left column, third paragraph). Therefore claim 360 is included in this rejection.
Seren teaches a PBS solution (supra). Therefore claim 361 is included in this rejection.
Seren teaches an aqueous medium comprising 0.05% Nonidet P-40 (octylphenoxypolyethoxyethanol). This reads on claims 365-369.
Seren isolates PR3 (see page 12425, right column, fourth paragraph). Therefore claim 370 is included in this rejection.
The method of claim 1 is rendered obvious. Kalupov teaches NE can be isolated (supra). Therefore claim 371 is included in this rejection.
Therefore Applicant’s Invention is rendered obvious as claimed.
Claims 362-364 are rejected under 35 U.S.C. 103 as being unpatentable over Seren in view of Kalupov as applied to claim 1 above, and further in view of Genaxxon Bioscience (Tris buffered saline (TBS) 2014, pages 1-2).
Claim 1 is rejected on the grounds set forth above. The teachings of the art are reiterated. Seren teaches an aqueous wash solution (phosphate buffered saline) comprising saline. The art does not teach the concentration of saline in PBS (claim 362). The art does not teach the use of Tris buffered saline (claims 363-364).
Genaxxon teaches Tris buffered saline (TBS) is isotonic and non-toxic to cells and is suitable for molecular biology. The buffer is commonly used as a wash buffer (see page 1, first paragraph of Product Description). Genaxxon teaches a TBS buffer comprising 0.05M Tris. 0.138M NaCl at a pH 8.0 (see Product specifications on page 1; 1X TBS pH 8.0). Because the claims do not define the term “about”, the amount of saline in TBS is interpreted to read on claim 362
It would have been obvious to use an aqueous wash comprising about 0.9% saline. Seren washes cells with saline solution and Genaxxon teaches washing cells with a solution comprising about 0.9% saline. One would do so since Genaxxon teaches it is non-toxic to cells. One would have had a reasonable expectation of success since Genaxxon teaches a solution comprising about 0.9% saline can be used to wash cells. One would have expected similar results since both references are directed to solutions for washing cells. Therefore claim 362 is rendered obvious.
It would have been obvious to use TBS comprising NaCl to wash cells. Seren washes cells and Genaxxon teaches washing cells with a TBS buffer. One would have been motivated to do so since Genaxxon teaches TBS is non-toxic to cells. One would have had a reasonable expectation of success since Genaxxon teaches TBS comprising NaCl can be used to wash cells. One would have expected similar results since both references are directed to solutions for washing cells. Therefore claim 363 is rendered obvious. Because the claims nor the specification define the term “about”, the concentrations and pH taught by Genaxxon are interpreted to read on claim 364.
Therefore Applicant’s Invention is rendered obvious as claimed.
Conclusion
No claims are allowed.
Any inquiry concerning this communication or earlier communications from the examiner should be directed to NATALIE MOSS whose telephone number is (571) 270-7439. The examiner can normally be reached on Monday-Friday, 8am-5pm EST.
If attempts to reach the examiner by telephone are unsuccessful, the examiner’s supervisor, Sharmila Landau can be reached on (571) 272-0614. The fax phone number for the organization where this application or proceeding is assigned is (571) 270-8439.
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/NATALIE M MOSS/ Examiner, Art Unit 1653