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 .
Continued Examination Under 37 CFR 1.114
A request for continued examination under 37 CFR 1.114, including the fee set forth in 37 CFR 1.17(e), was filed in this application after final rejection. Since this application is eligible for continued examination under 37 CFR 1.114, and the fee set forth in 37 CFR 1.17(e) has been timely paid, the finality of the previous Office action has been withdrawn pursuant to 37 CFR 1.114. Applicant's submission filed on 05/05/2026 has been entered.
Receipt is acknowledged of an amendment, filed on 05/05/2026, in which claims 8 and 9 were withdrawn in a previously established election of species, claims 1-7, 10, 13 and 14 are previously presented and claims 11 and 12 are amended.
Claims 1-7 and 10-14 are currently under examination.
Priority
Acknowledgment is made of applicant' s claim for priority based on a continuation application filed as PCT/US2021/058634 on 11/09/2021.
Acknowledgment is made of applicant' s claim for priority based on a provisional application filed as 63/113,817 on 11/13/2020.
All claims are given the priority date of 11/13/2020.
Claim Rejections - 35 USC § 102
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 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.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claims 1, 3-6 and 10-14 are rejected under 35 U.S.C. 102(a)(1)/(a)(2) as being anticipated by Shi et al (US 2018/0171295 A1; Cited in a Prior Office Action).
Regarding claim 1, Shi teaches a buoyancy enabled separation method for isolation from a sample including the sample cells 1 that are peripheral blood mononuclear cells (PBMC) 1 are mixed and incubated with anti-CDS microbubble 2 in a syringe-like container 3; Microbubbles 2 and targeted cells 4 gradually float to the liquid surface (FIG. 2); The targeted cell 4 population (CDS+ cells) is retained (positive selection) by pushing the plunger 5 (FIG. 3) with the container opening 6 facing the ground (FIG. 2); For the next cycle of BACS, the second type of targeted microbubble 2 (anti-CD62L) with buffer is mixed with the anti-CDS sorted cells; The process restarts as previously described (FIGS. 2-4); Therefore, after the 3rd cycle with anti-CD45RA microbubbles 2, the naive CDS+ T cell population (CDS+ CD62L+CD45RA+) in the syringe-like container 3 is stimulated and expanded by binding to anti-CD3/anti-CD2S conjugated microbubbles 2 in culture medium and subsequent manipulations ( e.g. viral transduction with chimeric antigen receptors) can also be performed in the same container 3 [0045] and (Page 2, Figs. 1-4). Shi teaches for CAR-T cell therapy, a CAR gene transduction can be performed within the container 3 by adding retroviral vectors with the cells and microbubbles within the incubating mixture [0070]. Shi teaches the microbubbles spontaneously burst within 10-16 hours in standard culture conditions at 37° C [0093].
Regarding claim 3, Shi teaches a buoyancy enabled separation method for isolation from a sample including the sample cells 1 that are peripheral blood mononuclear cells (PBMC) 1 are mixed and incubated with anti-CDS microbubble 2 in a syringe-like container 3; Microbubbles 2 and targeted cells 4 gradually float to the liquid surface (FIG. 2); The targeted cell 4 population (CDS+ cells) is retained (positive selection) by pushing the plunger 5 (FIG. 3) with the container opening 6 facing the ground (FIG. 2); For the next cycle of BACS, the second type of targeted microbubble 2 (anti-CD62L) with buffer is mixed with the anti-CDS sorted cells; The process restarts as previously described (FIGS. 2-4); Therefore, after the 3rd cycle with anti-CD45RA microbubbles 2, the naive CDS+ T cell population (CDS+ CD62L+CD45RA+) in the syringe-like container 3 is stimulated and expanded by binding to anti-CD3/anti-CD2S conjugated microbubbles 2 in culture medium and subsequent manipulations ( e.g. viral transduction with chimeric antigen receptors) can also be performed in the same container 3 [0045] and (Page 2, Figs. 1-4). Shi teaches for CAR-T cell therapy, a CAR gene transduction can be performed within the container 3 by adding retroviral vectors with the cells and microbubbles within the incubating mixture [0070].
Regarding claim 4, Shi teaches the microbubbles spontaneously burst within 10-16 hours in standard culture conditions at 37° C [0093].
Regarding claims 5 and 6, Shi teaches a buoyancy enabled separation method for isolation from a sample including the sample cells 1 that are peripheral blood mononuclear cells (PBMC) 1 are mixed and incubated with anti-CDS microbubble 2 in a syringe-like container 3; Microbubbles 2 and targeted cells 4 gradually float to the liquid surface (FIG. 2); The targeted cell 4 population (CDS+ cells) is retained (positive selection) by pushing the plunger 5 (FIG. 3) with the container opening 6 facing the ground (FIG. 2); For the next cycle of BACS, the second type of targeted microbubble 2 (anti-CD62L) with buffer is mixed with the anti-CDS sorted cells; The process restarts as previously described (FIGS. 2-4); Therefore, after the 3rd cycle with anti-CD45RA microbubbles 2, the naive CDS+ T cell population (CDS+ CD62L+CD45RA+) in the syringe-like container 3 is stimulated and expanded by binding to anti-CD3/anti-CD2S conjugated microbubbles 2 in culture medium and subsequent manipulations ( e.g. viral transduction with chimeric antigen receptors) can also be performed in the same container 3 [0045] and (Page 2, Figs. 1-4). Shi teaches for CAR-T cell therapy, a CAR gene transduction can be performed within the container 3 by adding retroviral vectors with the cells and microbubbles within the incubating mixture [0070].
Regarding claim 10, Shi teaches a buoyancy enabled separation method for isolation from a sample including the sample cells 1 that are peripheral blood mononuclear cells (PBMC) 1 are mixed and incubated with anti-CDS microbubble 2 in a syringe-like container 3; Microbubbles 2 and targeted cells 4 gradually float to the liquid surface (FIG. 2); The targeted cell 4 population (CDS+ cells) is retained (positive selection) by pushing the plunger 5 (FIG. 3) with the container opening 6 facing the ground (FIG. 2); For the next cycle of BACS, the second type of targeted microbubble 2 (anti-CD62L) with buffer is mixed with the anti-CDS sorted cells; The process restarts as previously described (FIGS. 2-4); Therefore, after the 3rd cycle with anti-CD45RA microbubbles 2, the naive CDS+ T cell population (CDS+ CD62L+CD45RA+) in the syringe-like container 3 is stimulated and expanded by binding to anti-CD3/anti-CD2S conjugated microbubbles 2 in culture medium and subsequent manipulations ( e.g. viral transduction with chimeric antigen receptors) can also be performed in the same container 3 [0045] and (Page 2, Figs. 1-4).
Regarding claim 11, Shi teaches a buoyancy enabled separation method for isolation from a sample including the sample cells 1 that are peripheral blood mononuclear cells (PBMC) 1 are mixed and incubated with anti-CDS microbubble 2 in a syringe-like container 3; Microbubbles 2 and targeted cells 4 gradually float to the liquid surface (FIG. 2); The targeted cell 4 population (CDS+ cells) is retained (positive selection) by pushing the plunger 5 (FIG. 3) with the container opening 6 facing the ground (FIG. 2); For the next cycle of BACS, the second type of targeted microbubble 2 (anti-CD62L) with buffer is mixed with the anti-CDS sorted cells; The process restarts as previously described (FIGS. 2-4); Therefore, after the 3rd cycle with anti-CD45RA microbubbles 2, the naive CDS+ T cell population (CDS+ CD62L+CD45RA+) in the syringe-like container 3 is stimulated and expanded by binding to anti-CD3/anti-CD2S conjugated microbubbles 2 in culture medium and subsequent manipulations ( e.g. viral transduction with chimeric antigen receptors) can also be performed in the same container 3 [0045] and (Page 2, Figs. 1-4). Shi teaches for CAR-T cell therapy, a CAR gene transduction can be performed within the container 3 by adding retroviral vectors with the cells and microbubbles within the incubating mixture [0070].
Regarding claims 12 and 13, Shi teaches CD3/CD28 affinity ligand combination conjugated microbubbles are infused for the isolation, sorting, expansion, manipulation, or stimulation of T cells [0059]. Shi teaches T cell expansion by using anti-CD3/CD2S conjugated microbubbles for one or more re-stimulations with different blood donors [0039].
Regarding claim 14, Shi teaches for CAR-T cell therapy, a CAR gene transduction can be performed within the container 3 by adding retroviral vectors with the cells and microbubbles within the incubating mixture wherein the microbubbles are binding to CD 19 cell surface markers CD 19 expressed T-cells [0070].
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 text of those sections of Title 35, U.S. Code not included in this action can be found in a prior Office action.
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.
Claim 2 is rejected under 35 U.S.C. 103 as being unpatentable over Shi et al (US 2018/0171295 A1; Cited in a Prior Office Action) in view of Taylor et al (J Gene Med 2007; 9: 77–87.).
The teachings of Shi are as described and applied above.
Regarding claim 2, Shi does not teach wherein the viruses, viral vectors or virus-like particles are first bound to the microbubbles for concentrating these on the microbubbles before the target cells are introduced into the mixture for binding subsequently to the microbubbles with the viruses, viral vectors or virus-like particles already bound to the microbubbles.
Taylor teaches envelope-deficient retroviral vector, inherently incapable of target cell entry, was combined with cationic microbubbles and subsequently added to target cells in order to complete successful viral transduction to the target cells (Page 77, Methods and Results).
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 teachings of Shi to include the virus or virus-like particles are first bound to the microbubbles before the target cells are introduced as taught by Coelho because Shi teaches it is within
the ordinary skill in the art to use a method for ex vivo transduction of biomolecules from viruses, viral vectors or virus-like particles into target cells, comprising: preparing a mixture by mixing a quantity of viruses, viral vectors or virus- like particles and flexible lipid shell microbubbles, said flexible lipid shell microbubbles being conjugated with one or more ligands binding to the viruses, viral vectors or virus-like particles and to the target cells; incubating the mixture over a time span allowing the viruses, viral vectors or virus-like particles to bind to microbubbles; incubating the microbubbles with the viruses, viral vectors or virus-like particles and the target cells to allow transduction to take place, transferring the biomolecules from viruses, viral vectors or virus-like particles into the target cells while the viruses, viral vectors or virus-like particles and the target cells are bound to the microbubbles and Taylor teaches conjugating the retroviral vectors to microbubbles first before introduction to the target cells always for successful viral transduction.
One would have been motivated to make such a modification in order to receive the expected benefit of successful viral transduction as taught by Taylor.
Claim 7 is rejected under 35 U.S.C. 103 as being unpatentable over Shi et al (US 2018/0171295 A1; Cited in a Prior Office Action) in view of Ji et al (US 2004/0028654 Al; Cited in a Prior Office Action) and Wang et al (Biomaterials, 183, pgs. 173-184; 2018; Cited in a Prior Office Action).
The teachings of Shi are as described and applied above.
Regarding claim 7, Shi does not teach wherein the target cells are T cells and the viral vectors are retroviral vectors, and the microbubbles are conjugated with protamine that binds the viral vectors and an RGD peptide that binds the target cells.
Wang teaches the development of a novel conjugate complex of plectin/integrin-targeted
bispecific molecular probe, termed Gd-Cy7-PTP/RGD (Page 173, Abstract). Wang teaches the bispecific molecular probe comprises four parts: Gd (III) for MRI, cyanine 7 (Cy7) for NIRF, the
peptide PTP for binding to plectin-1 specifically overexpressed on the surface of PDAC cells,
and the peptide RGD for targeting integrin widely expressed on pancreatic duct epithelial cells
and angiogenesis (Page 174, Column 1).
Wang does not teach the use of the bispecific ligand to bind the viral vectors, viruses or
viral-like particles.
Ji teaches Protamine is a natural, arginine-rich peptide with an overall positive charge
[0013]. Ji teaches methods and compositions of the invention involve a viral composition that
can be administered systemically wherein the invention includes viral compositions having
improved transduction efficiency in vitro, ex vivo, and in vivo and the methods provide for an
increased transduction efficiency and therapeutic efficacy in cancer cells and tumors, in
particular cancer and tumor cells associated with the lung [0012]. Ji teaches the invention
includes viral compositions comprising a (a) a protamine molecule and (b) a therapeutic viral
vector [0012]. Ji teaches a viral vector and protamine molecule complex can be used for
increasing transduction efficiencies, increasing therapeutic efficacy and alleviating side effects of
viral vector therapy, such as neutralizing antibody production and hepatic toxicity [0014]. Ji
teaches the targeting ligand may be operably coupled to a protamine molecule either directly,
e.g. fusion protein, or indirectly by means of linking moiety, for example as a bispecific ligand
including protamine and a targeting peptide [0016]. Ji teaches the targeting moiety is a ligand,
such as a peptide ligand that interacts with cell surface receptors, such as EGFR, VEGFR, and
CAR [0015].
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 teachings of Shi to include the microbubbles are conjugated with a ligand binding to a virus for transduction to a target cell as taught by Ji and the
RGD peptide for specific cell targeting as taught by Wang because Shi teaches it is within
the ordinary skill in the art to use a method for ex vivo transduction of biomolecules from viruses, viral vectors or virus-like particles into target cells, comprising: preparing a mixture by mixing a quantity of viruses, viral vectors or virus- like particles and flexible lipid shell microbubbles, said flexible lipid shell microbubbles being conjugated with one or more ligands binding to the viruses, viral vectors or virus-like particles and to the target cells; incubating the mixture over a time span allowing the viruses, viral vectors or virus-like particles to bind to microbubbles; incubating the microbubbles with the viruses, viral vectors or virus-like particles and the target cells to allow transduction to take place, transferring the biomolecules from viruses, viral vectors or virus-like particles into the target cells while the viruses, viral vectors or virus-like particles and the target cells are bound to the microbubbles, Wang teaches the development of a novel conjugate complex of plectin/integrin-targeted bispecific molecular probe, termed Gd-Cy7-PTP/RGD and Ji teaches using protamine in a bispecific targeting ligand such as using protamine to bind a virus and a targeting peptide to target cell surface receptors such as CAR.
One would have been motivated to make such a modification in order to receive the
expected benefit of a protamine ligand for improved transduction of viral particles to a target cell
as taught by Ji and a bispecific ligand utilizing an RGD peptide to bind to target cells as taught
by Wang.
Claim Rejections - 35 USC § 103
The previous rejection of claims 1, 2, 4, 6 and 10 under 35 U.S.C. 103 as being unpatentable over Claudio et al (US 2008/0063604 Al) in view of Ji et al (US 2004/0028654 Al) has been withdrawn.
The previous rejection of claim 7 under 35 U.S.C. 103 as being unpatentable over Claudio et al (US 2008/0063604 Al) in view of Ji et al (US 2004/0028654 Al) and further in view of Wang et al (Biomaterials,183, pgs. 173-184; 2018) has been withdrawn.
The previous rejection of claims 3, 5, 11, 12, 13 and 14 under 35 U.S.C. 103 as being unpatentable over Claudio et al (US 2008/0063604 Al) in view of Ji et al (US 2004/0028654 Al) and further in view of Shi et al (US 2018/0171295 Al) has been withdrawn.
Conclusion
No claims are allowed.
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/ALEXANDRA ROSE LIPPOLIS/Examiner, Art Unit 1637
/CELINE X QIAN/Primary Examiner, Art Unit 1637
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