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 .
Status of the Claims
Claims 1-2, 5, 7, 9, 14-19, 21, 23, 27-28, 49-50, 53, 55, and 57 are currently pending (claim set as filed on 9/27/2024). Claims 3-4, 6, 8, 10-13, 20, 22, 24-26, 29-48, 51-52, 54, 56, and 58-61 are cancelled. Claims 49-50, 53, 55, and 57 are withdrawn due to a restriction/election requirement. Claims 1-2, 5, 7, 9, 14-19, 21, 23, and 27-28 are under examination.
Election/Restrictions
Claims 49-50, 53, 55, and 57 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to a nonelected product, there being no allowable generic or linking claim. Election was made without traverse in the reply filed on 8/31/2026. Applicant’s election without traverse of the species elections of targeting ligand, sample type, and microbial cell in the reply filed on 8/31/2026 is acknowledged.
Priority
Acknowledgement is made of applicant’s claim to provisional application no. 63/241,214, filed on 9/7/2021. Thus the effective filing date of this application is 9/7/2021.
Information Disclosure Statement
The information disclosure statements (IDS) submitted on 3/7/2024, 5/10/2024, and 9/25/2025 were considered, initialed, and attached hereto. The submissions are in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statements are being considered by the examiner.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
Claims 1-2, 5, 7, 9, 14-19, 21, 23, and 27-28 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Regarding claim 1, the phrase “wherein the particle comprises a core and a shell” renders the claim indefinite as it is unclear what part of the particle structure has a core and shell; the nanodroplet/microbubble, the targeting ligand, both, or the conjugate structure as a whole. For examination purposes, the language is interpreted to mean the nanodroplet/microbubble comprises a core and a shell and the prior art shall be applied accordingly. Additionally, the term “low-pressure sonication” in claim 1 is a relative term which renders the claim indefinite. The term is not defined by the claim, the specification does not provide a scientific standard for ascertaining the requisite degree and its metes and bounds, and one of ordinary skill in the art would not be reasonably apprised of the scope of the invention. Although embodiments are provided in the instant specification, it is unknown as to what frequencies are intended to be included within the term or alternatively excluded from the term.
Claims 2, 5, 7, 9, and 14-19 are rejected for depending on claim 1.
Regarding claims 2, 5, 7, 9, and 23, the phrase "e.g." renders the claims indefinite because it is unclear whether the limitation(s) of:
“concanavalin A, wheat germ agglutinin, or mannose/mannan binding protein” and “an anti-lipoarabinomannan antibody, an anti-protein A antibody, a lipoteichoic acid antibody, or an anti GM 1 antibody” in claim 2
“by click chemistry, thiol/maleimide, thiol/thiol, hydrazide/aldehyde, gold/thiol, or biotin/streptavidin-based techniques” in claim 5
“a lipid” and “wherein the shell comprises 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), polyethylene glycol (PEG), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), dibenzocyclooctyne (DBCO), 1,2- distearoyl-sn-glycero-3-phosphoethanolamine-N-rmethoxy(polyethylene glycol)-2000] (DSPE-PEG2000), another DSPE with another PEG chain length, and/or DBCO with a click chemistry linker” in claim 7
“wherein the core comprises a gas with a boiling point of -100 *C to 50 °C, e.g., wherein the core comprises a perfluorocarbon, e.g., wherein the perfluorocarbon is octofluoropropane, decafluorobutane, or hexafluoropentane, e.g., wherein the core comprises sulfur hexafluoride” in claim 9
“wherein the floating is performed naturally without any additional steps or wherein the floating is performed by centrifuging the sample, e.g., wherein the centrifuging is performed in a syringe, plugged/capped pipette tip, capillary tube, or centrifuge tube” in claim 23
following the phrase are part of the claimed invention. See MPEP § 2173.05(d).
Regarding claims 2, 5, and 7, a broad range or limitation together with a narrow range or limitation that falls within the broad range or limitation (in the same claim) may be considered indefinite if the resulting claim does not clearly set forth the metes and bounds of the patent protection desired. See MPEP § 2173.05(c). In the present instance, the claims recite the broad recitations of:
“wherein the targeting ligand is a lectin” and “an antibody” in claim 2
“wherein the targeting ligand is conjugated to the shell of the particle” in claim 5
“wherein the shell comprises a surfactant” and “a combination thereof” in claim 7
and the claims also respectively recite:
“concanavalin A, wheat germ agglutinin, or mannose/mannan binding protein” and “an anti-lipoarabinomannan antibody, an anti-protein A antibody, a lipoteichoic acid antibody, or an anti GM 1 antibody” in claim 2
“by click chemistry, thiol/maleimide, thiol/thiol, hydrazide/aldehyde, gold/thiol, or biotin/streptavidin-based techniques” in claim 5
“a lipid” and “wherein the shell comprises 1, 2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), polyethylene glycol (PEG), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2- Page 2 of 7 dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), dibenzocyclooctyne (DBCO), 1,2- distearoyl-sn-glycero-3-phosphoethanolamine-N-rmethoxy(polyethylene glycol)-2000] (DSPE-PEG2000), another DSPE with another PEG chain length, and/or DBCO with a click chemistry linker” in claim 7
which is the narrower statement of the range/limitation. The claims are considered indefinite because there is a question or doubt as to whether the feature introduced by such narrower language is (a) merely exemplary of the remainder of the claim, and therefore not required, or (b) a required feature of the claims.
Regarding claim 21, the claim recites “and floating the microbial cell bound to the particle to the top of the sample” which renders the claim indefinite. The claims state that the sample comprises the microbial cell with no mention in regard to other components of the sample. Thus, if the sample merely comprises the microbial cell it is unclear as to what additional element of the sample the microbial cell-particle conjugate is floating towards. For examination purposes, the language has been broadly interpreted to mean the microbial cell-particle conjugate produces an inverse pellet.
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.
Claims 1-2, 5, 7, 9, 14, 16-19, 21, 23, and 27-28 are rejected under 35 U.S.C. 103 as being unpatentable over Lattwein (Lattwein et al., “Sonobactericide: an emerging treatment strategy for bacterial infections”, 2019 Sep 16, Ultrasound in Medicine and Biology, 46(2), pgs. 193-215) and as evidenced by Akadeum (Akadeum Life Sciences, “The Physics of How Microbubbles Sort Cells”, 2021 Jun 23, www.akadeum.com/technology/the-physics-of-how-microbubbles-sort-cells/?srsltid=AU7gw4Vbq063Kxs8yTZAadxQ2i2_JKCUA2lu6ipfg7To8kghqTbs501c, pgs. 1-4, accessed 2026 Sep 25).
Lattwein’s general disclosure relates to a sonobactericide method wherein targeted microbubbles are exposed to ultrasonic frequencies in order to disrupt bacterial biofilms (see abstract).
Regarding claim 1, Lattwein teaches a microbubble particle that comprises a shell and a core and is bound to a bacterial cell surface (see Lattwein pg. 198 – Figure 2a).
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Lattwein teaches microbubbles can be further modified by the addition of ligands in the coating (see Lattwein pg. 204, ¶ 3). Lattwein discloses cavitating the microbubbles on the bacterial biofilm surfaces, which suggests contact with the cell wall, via ultrasound vibrations to disrupt the biofilm (see Lattwein pg. 206, ¶ 6). The instant specification recites “nanodroplets enable efficient use of ultrasound for the lysis of microbes” (see specification [0048]), and thus the use of ultrasound would read on the claimed limitation of “sonication” as they have the same purpose and function. Lattwein further discloses that after ultrasonic treatment, the bacterial cells were lysed and intracellular DNA was released into the supernatant (see Lattwein pg. 202, ¶ 3 and pg. 203, ¶ 2). As the instant specification states the biomolecules can refer to “any molecule that can be obtained from a cell, including without limitation, DNA, RNA, intracellular protein, intramembrane/cell wall protein, and metabolites” (see specification [0029]), the release of DNA would read on this limitation.
However, Lattwein does not teach the method components in one singular embodiment and one would not immediately envisage the combination as claimed.
It would be obvious to one of ordinary skill in the art at the time of the effective filing date of the invention to combine the method steps of contacting the microbubble that is functionalized with a ligand to a microbial cell and applying ultrasonic vibrations to the microbubble/microbe sample to extract biomolecules like DNA. The ordinary artisan would have been motivated to do so as Lattwein discloses that the concepts of microbubble composition, ultrasound parameters, and bacteria are the main principles of sonobactericide therapy (see pg. 194, ¶ 2). Hence, the ordinary artisan would have found it obvious to combine into one singular embodiment as these are the foundational concepts of the art and work together to arrive at the end result of disrupting bacterial biofilms.
Regarding claim 2, Lattwein teaches a targeting ligand, such as anti-protein A antibodies, can be added to the microbubbles (see pg. 204, ¶ 3-4).
Regarding claim 5, Lattwein teaches the ligand is attached, which is conjugated, to the microbubble via the coating on the surface of the microbubble (see pg. 204, ¶ 3).
Regarding claims 7 and 9, Lattwein teaches the microbubble surface coating comprises a surfactant and the core is gaseous air (see pg. 204, ¶ 2).
Regarding claim 14, Lattwein teaches the ultrasound vibrations were administered at a frequency of 1 MHz (see pg. 206, ¶ 6) which is within the claimed range of about 1 kHz to about 100 MHz.
Regarding claim 16, Lattwein teaches the concentration of the microbubble particles is between 2 x 108 to 5 x 108 microbubbles/mL (see pg. 201, ¶ 4), which is fully encompassed within about 1x102 to about 1x1012 particles per ml.
Regarding claim 17, Lattwein teaches the samples are patient-derived clinical isolates (see pg. 195, ¶ 4).
Regarding claim 18, Lattwein teaches the microbial cells are Gram-positive bacteria (see pg. 198 – Figure 2a).
Regarding claim 19, Lattwein teaches the microbial cells are Gram-positive bacteria (see pg. 198 – Figure 2a) which are microbes and that they also form biofilms (see pg. 204, ¶ 4).
Regarding claim 21, Lattwein teaches a microbubble particle that comprises a shell and a core and is bound to a bacterial cell surface (see Lattwein pg. 198 – Figure 2a). Lattwein teaches microbubbles can be further modified by the addition of ligands in the coating (see Lattwein pg. 204, ¶ 3). As noted in the 112b rejection above, it is unclear as to how the microbial cell-particle conjugate can float to the top of the sample if the sample merely comprises the microbial cell itself. For examination purposes, the language has been broadly interpreted to mean the microbial cell-particle conjugate produces an inverse pellet. Lattwein also teaches the post-treatment samples can be centrifuged (see Lattwein pg. 202, ¶ 2). As evidenced by Akadeum, microbubbles have a low density and naturally gravity causes them to rise to the top of the container forming an inverse pellet (see Akadeum pg. 2 – “Why do Microbubbles Float?”). As the microbubbles are attached to the microbial cells, the microbial cells would also naturally comprise the pellet. Since the prior art and evidentiary reference teach the same structure of a microbial cell bound to a microbubble particle that can form an inverse pellet as the instant application, it would naturally have the functional limitation of “selectively enriching the microbial cell” as well.
Regarding claim 23, Lattwein teaches the particle can be a nanodroplet and be in contact with Gram-negative bacteria (see pg. 198 – Figure 2b).
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Lattwein teaches the nanodroplet-microbial cell conjugate can be subject to ultrasound vibrations that phase transition the nanodroplet into microbubbles (see pg. 205, ¶ 2). Lattwein teaches the bacterial biofilms can be centrifuged, and naturally resulting in an inverted pellet, post-treatment (see pg. 202, ¶ 2). Lattwein discloses that the ultrasound treatment can, but seemingly does not require, inertial cavitation to be performed instead of stable cavitation (see pg. 206, ¶ 2). Thus it would be obvious to the ordinary artisan to utilize a lower pressure cavitation method instead as the prior art suggests to suit their needs.
Regarding claim 27, Lattwein teaches the attachment of the microbubbles to the microbial cells (see Lattwein pg. 198 – Figure 2a) and as evidenced by Akadeum, the microbubbles will naturally float the surface of their container as an inverse pellet by gravity (see Akadeum pg. 2 – “Why do Microbubbles Float?”). Lattwein further teaches the biofilm post-treatment can be vortexed (see Lattwein pg. 202, ¶ 2) which would resuspend the inverse pellet and the microbes within it.
Regarding claim 28, Lattwein teaches the biofilm cells can undergo centrifugation post-treatment (see Lattwein pg. 202, ¶ 2) and naturally the cells would form inverse pellets due to their attachment to microbubbles, thus enriching the cells. As centrifugation results in the formation of a supernatant, the DNA extracts of the spun biofilm cells would be present in such a substance. The instant specification states the biomolecules can refer to “any molecule that can be obtained from a cell, including without limitation, DNA, RNA, intracellular protein, intramembrane/cell wall protein, and metabolites” (see specification [0029]), and so the release of DNA would read on this limitation.
Claim 15 is rejected under 35 U.S.C. 103 as being unpatentable over Lattwein (Lattwein et al., “ Sonobactericide: an emerging treatment strategy for bacterial infections”, 2019 Sep 16, Ultrasound in Medicine and Biology, 46(2), pgs. 193-215), further in view of Keravnou (Keravnou et al., “Accurate Measurement of Microbubble Response to Ultrasound With a Diagnostic Ultrasound Scanner”, 2014 Oct 3, IEE Transactions on Ultrasonics, Ferroelectrics, and Frequency Control, 62(1), pgs. 176-184), and as evidenced by Akadeum (Akadeum Life Sciences, “The Physics of How Microbubbles Sort Cells”, 2021 Jun 23, www.akadeum.com/technology/the-physics-of-how-microbubbles-sort-cells/?srsltid=AU7gw4Vbq063Kxs8yTZAadxQ2i2_JKCUA2lu6ipfg7To8kghqTbs501c, pgs. 1-4, accessed 2026 Sep 25).
Regarding claim 15, Lattwein teaches a microbubble particle and a microbial cell sample (see pg. 198 – Figure 2a).
However, Lattwein does not teach the particle to microbial cell ratio was 1:10 v/v.
Keravnou’s general disclosure relates to the phenomena of acoustic shadowing that occurs when microbubble concentration is high, limiting the depth of ultrasound vibrations to the affected area (see abstract).
Regarding claim 15, Keravnou teaches a solution with microbubbles at a low concentration (see pg. 177, ¶ 4).
It would have been obvious to one of ordinary skill in the art before the effective filing date of the invention to optimize the ratio of microbubbles at a lower concentration as taught in Keravnou to the 1:10 v/v ratio as claimed. The ordinary artisan would have been motivated to do so because Keravnou teaches that higher densities of microbubbles in a solution can actually impede ultrasound functioning by limiting acoustic pressure into deeper areas (see Keravnou abstract). This teaching would have been an advantage to Lattwein’s disclosure which also recognizes that increasing microbubble density can dampen ultrasound oscillations (see Lattwein pg. 205, ¶ 5). Thus by diluting the concentration of microbubbles to 1:10 compared to the microbial cells in the sample, one could improve the ultrasound frequency needed to extract biomolecules from the microbial cells present.
Conclusion
No claims are allowed.
Correspondence Information
Any inquiry concerning this communication or earlier communications from the examiner should be directed to Emmalee R. Williams whose telephone number is (571)272-5472. The examiner can normally be reached Monday - Friday 7:30 am - 5:00 pm.
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/EMMALEE R WILLIAMS/ Examiner, Art Unit 1653
/JENNIFER M.H. TICHY/Primary Examiner, Art Unit 1653
