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 .
Claims 15, 17, 19, 22-30, 63-66, and 85 have been canceled. Claims 1, 6-14, 16, 18, 20, 21, 31, 32, 40, 47, 48-58, 60, 62, 67, 68, 70, 75-77, 79-84 and 86 have been amended. Claims 89-91 have been added. Claims 1-14, 16, 18, 20, 21, 31-62, 67-84 and 89-91 are pending and under consideration.
The objection to claims 10-14, 16, 18, 20, 21, 31-62, 67-84, and 86 under 37 CFR 1.75(c) as being in improper form because a multiple dependent claim cannot serve as the basis for another multiple dependent claim is withdrawn in light of applicant’s amendments.
The objection to claim 7 for a typographical error is withdrawn in light of applicant’s amendment.
Claim 75 is objected to because of the following informalities: reference to section d) which has been canceled.. Appropriate correction is required.
Applicant’s claim for the benefit of a prior-filed application under 35 U.S.C. 119(e) or under 35 U.S.C. 120, 121, 365(c), or 386(c) is acknowledged. Applicant has not complied with one or more conditions for receiving the benefit of an earlier filing date under 35 U.S.C. 119(e) as follows:
The later-filed application must be an application for a patent for an invention which is also disclosed in the prior application (the parent or original nonprovisional application or provisional application). The disclosure of the invention in the parent application and in the later-filed application must be sufficient to comply with the requirements of 35 U.S.C. 112(a) or the first paragraph of pre-AIA 35 U.S.C. 112, except for the best mode requirement. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
The disclosure of the prior-filed applications, Application No.63/068,249 and 63/105,639, fail to provide adequate support or enablement in the manner provided by 35 U.S.C. 112(a) or pre-AIA 35 U.S.C. 112, first paragraph for one or more claims of this application. Claim 32 requires the immune cell of claim 31 wherein the cell is modified to reduce or eliminate expression of the B2M gene product. Claims 33 and 34 require the immune cell of claim 32 comprising the sequence complementary to B2M RNA of SEQ ID NO: 172. Claim 35 and 36 require that the polynucleotide of claim 33 and 35 comprise a sequence encoding a shRNA specific to B2M. Claim 37specifies that the on or mor modifications ofB2N is one or more modifications to SEQ ID NO: 170. Clams 38-40 require that the one or more modifications of claims 37 comprise one or more inactivating mutations of the endogenous gene encoding B2M, that the one or more inactivating mutations comprise a deletion, insertion or frameshift mutations and wherein the one or more mutations are introduced with a gNA that specifically targets the endogenous SEQ ID NO: 170. Neither 63/068,249, nor 63/105,639 provide a written description of an immune cell modified to reduce or eliminate expression of the B2M gene product or a shRNA specific to B2M. Accordingly, the effective priority date for claims 32-40 will be the date of the first description of the inventive cell and polynucleotide having said characteristics as disclosed in 63/230,632, filed on 08/06/2021.
The following is a quotation of 35 U.S.C. 112(d):
(d) REFERENCE IN DEPENDENT FORMS.—Subject to subsection (e), a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
The following is a quotation of pre-AIA 35 U.S.C. 112, fourth paragraph:
Subject to the following paragraph [i.e., the fifth paragraph of pre-AIA 35 U.S.C. 112], a claim in dependent form shall contain a reference to a claim previously set forth and then specify a further limitation of the subject matter claimed. A claim in dependent form shall be construed to incorporate by reference all the limitations of the claim to which it refers.
Claims 18, 21, 34 , 39, 59-62 and 76-84 are rejected under 35 U.S.C. 112(d) or pre-AIA 35 U.S.C. 112, 4th paragraph, as being of improper dependent form for failing to further limit the subject matter of the claim upon which it depends, or for failing to include all the limitations of the claim upon which it depends.
(A)Claim 18 states “functional variant thereof” in reference to the LILRB1 transmembrane domain of the immune cell of claim 1. Claim 1 requires that the second receptor comprises a LILRB1 intracellular domain, a LILRB1 transmembrane domain and a LILRB1 hinge domain. Thus claim 18 as drawn to the “functional variant “ of the “LILRB1 transmembrane domain fails to include all the limitation of claim 1 which is limited to LILRB1 transmembrane domain and not a functional variant.
(B)Claim 21 states “functional variant thereof” in reference to the LILRB1 transmembrane domain of the immune cell of claim 20. Claim 20 is dependent on claim 1 requiring that the second receptor comprises a LILRB1 intracellular domain, a LILRB1 transmembrane domain and a LILRB1 hinge domain. Thus claim 21 as drawn to the “functional variant “ of the “LILRB1 transmembrane domain fails to include all the limitation of the claim on which it depends.
(C)Claim 39 states that one or more inactivating mutations comprise a deletion, insertion, substitution or a frameshift mutation of the immune cell of claim 38. Claim 38 requires one or more inactivating mutations of the endogenous gene encoding B2M. Stating all of the possibilities for obtaining an inactivating mutation in claim 39 fails to further limit claim 38.
(D) claims 59 states that the immune cell of the pharmaceutical composition of claim 58, expresses both of the first and second receptor. Claim 58 is ultimately dependent on claim 1 which requires that the immune cell expresses both of a first receptor and a second receptor. Thus, claim 59 stating the same, fails to further limit the claim on which it is dependent. .
(E)Claims 60 states that the immune cells of claim 59, wherein about 50% to 90% of the immune cells express both the first and second receptor. Claim 61 states that at least 90% of the immune cells express both of the first and second receptor. Claims 60 and 61 ultimately dependent on claim 1 which requires that the immune cell expresses both of a first receptor and a second receptor. Thus, claims 60 and 61 fail to include all the limitations of the claim on which they depend.
(F)Claims 76-84:
Claim 76 states that the administration of an effective amount of the immune cell to a subject with cancer reduces the size of a tumor in the subject. in the method of claim 75
Claim 77 states that the tumor size is reduced by about 5%. in the method of claim 76.
Claim 78 states that the tumor s eliminated in the method of claim 76..
Claim 79 states that the administration of an effective amount of the immune cell to a subject with cancer arrests the growth of a tumor in the subject in the method of claim 75.
Claim 80 states that the administration of the immune cell results in the reduction of the number of tumors in the subject in the method of claim 75.
Claim 81 states that the administration if the immune cell results in the selective killing of a cancer cell but not a normal cell in the subject in the method of claim 80.
Claim 82 state that at least 60% of the cell skilled are cancer cells in the method of claim 81.
Claim 83 states that the administration of the immune cells results in the killing of about 40% of the cancer cells of the subject in the method of claim 81.
None of the methods of claims 76-83 further limit the subject matter of the claim on which they depend. All of claims 76-83 are ultimately dependent on claim 75. Claim 75 requires a) determining the genotype or expression level of a non-target antigen in non-malignant cells and cancer cells of the subject; b) determining the expression vel of EGFR in cancer cell of the subject and c) administering to the subject an effective amount of the immune cell of claim 88 if the non-malignant cells express the non-target antigen and the cancer cells are EGFR+ and do not express the non-target antigen,. None of claims 76-83 change the scope of the method or add to the method steps. It is noted that a “wherein” clause in a method claim is not given weight when it simply expresses the intended result of a process step positively recited, see MPEP 2111.04. It is noted that the observed results in claims 76-83 are the intended results of a process step positively recited. The particular outcomes of claims 76-83 would be a function of the particular characteristics of the tumor cells of the subject including molecular and physiological differences in the tumor tissue, tumor burden in the subject, and the individual characteristics of the subject’s immune system. However, in all claims 76-83 the actual method steps remain the same as claim 75. Thus, claims 76-83 fails to further limit the subject matter of the claim upon which it depends.
Claim 84 states that the administration of the immune cell results in fewer side effects for the subject than administration of an otherwise equivalent immune cell comprising the first receptor without the second inhibitory receptor. The fewer side effects of the administered immune cell is an inherent characteristic of the inventive immune cell, and the method of claim 75 is unchanged. Thus, claim 84 fails to further limit the subject matter of the claim upon which it depends.
(G) Claim 34 states that the interfering RNA of claim 33 is capable of RNAi-mediated degradation of B2M RNA. However, claim 33 requires interfering RNA which comprises a sequence complementary to the B2M RNA of SEQ ID NO: 172. Interfering RNA, by definition, is capable of inducing degradation of the target mRNA. Thus, claim 34 fails to further limit claim 33.
Applicant may cancel the claim(s), amend the claim(s) to place the claim(s) in proper dependent form, rewrite the claim(s) in independent form, or present a sufficient showing that the dependent claim(s) complies with the statutory requirements.
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 11-13 and 31-47, 67-84, 88-91 are rejected and claims 6 and 8 remain 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.
(A)Claim 11 is vague and indefinite in the recitation of “Table 3”as a means for identifying the heavy chain CDRs of the extracellular ligand binding domain of the first receptor. Claim 12 is vague and indefinite in the recitation of “Table 2” as a means for identifying the heavy chain variable regions of the extracellular ligand binding domains of claim 11. Claim 13 is vague and indefinite in the recitation of “Table 1” as a means for identifying the sequences of the extracellular ligand binding domain of the first receptor. Claims 6 is vague and indefinite in the recitation of “Table 5” as a means for identifying the CDR sequences of the second receptor. Claim 8 is vague and indefinite in the recitation of “Table 4” as a means for identifying the extracellular binding domains of the second receptor.
Section 2173.05(s) of the M.P.E.P. states:
Reference to Figures or Tables
Where possible, claims are to be complete in themselves. Incorporation by reference to a specific figure or table “is permitted only in exceptional circumstances where there is no practical way to define the invention in words and where it is more concise to incorporate by reference than duplicating a drawing or table into the claim. Incorporation by reference is a necessity doctrine, not for applicant’s convenience.” Ex parteFressola, 27 USPQ2d 1608, 1609 (Bd. Pat. App. & Inter. 1993).
Thus, claims 6, 8 and 11-13 are vague and indefinite because they are not complete in themselves.
Applicant argues that this is a case of exceptional circumstances wherein there is no practical way to define the invention in words. This has been considered but not found persuasive. If the sequences of Tables 1-5 can be described in the specification by SEQ ID NO, they can be described in the claims by SEQ ID Nos.
(B)Claim 11 is vague and indefinite in the recitation of “or CDR sequences having at most 1, 2, 3, 4 substitutions, insertions or deletions in each CDR” because it is unclear if this pertains to the heavy chain CDRs, of Table 3, the light chain CDRs of Table 3, both of the heavy and light chain CDRs of Table 3, or other CDR sequences.
(C) Claim 12 is vague and indefinite because it is unclear if sequences having at least 70%, at least 80%, at least 90% or at least 95% identity thereto pertains to the Vl portion, the Vh portion or both of the Vl and Vh portions.
(D)Claim 31 and claims 32-62, 67-84, 88-91, dependent on claim 31, are vague and indefinite in reliance on canceled claim 22. For purpose of examination, claim 31 will be read as dependent on claim 21.
(E)It is unclear how the method for introducing the inactivating mutations in claims 40 and 47 alters the scope of the immune cells of claims 39 and 46, which are required to have one or more inactivating mutations in the endogenous genes encoding B2M and HLA-A*02, respectively.
(F) Claims 37 and 38 are vague and indefinite in the recitation of “one or more modification to a sequence encoding B2M” because it is unclear if applicant considers the introns of the gene encoding B2M to be part of the sequence encoding B2M.
(G) Claims 45 and 46 are vague and indefinite in the recitation of “one or more modification to a sequence encoding HLA-A*02” because it is unclear if applicant considers the introns of the gene encoding HLA-A*02 to be part of the sequence encoding B2M.
(H)Claim 75 is vague and indefinite in the recitation of “determining the…expression level of a non-target antigen” (part a) and “determining the expression level of EGFR” (part b) followed by administering to the subject an effect amount of the immune cells of claim 88 if the non-malignant cells express the target antigen and the cancer cells do not express the non-target antigen, and the cancer cells are EGFR positive (part c) The claim is unclear because part c does not require an action based on “expression level”. Rather part c requires only that the cancer cells are EGFR positive and the non-malignant cells express the non-target antigen and the cancer cells do not express the non-target antigen. Thus, part c does not utilize the expression levels determined in parts a and b but only requires the presence or absence of the of EGFR and the non-target antigen. It is unclear what applicant intends to use the determined expression levels for.
The following is a quotation of the first paragraph of 35 U.S.C. 112(a):
(a) IN GENERAL.—The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor or joint inventor of carrying out the invention.
The following is a quotation of the first paragraph of pre-AIA 35 U.S.C. 112:
The specification shall contain a written description of the invention, and of the manner and process of making and using it, in such full, clear, concise, and exact terms as to enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make and use the same, and shall set forth the best mode contemplated by the inventor of carrying out his invention.
Claims 11-14, 16, 18, 20, 21, 31-62, 67-84 and 86-91 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, as failing to comply with the written description requirement. The claim(s) contains subject matter which was not described in the specification in such a way as to reasonably convey to one skilled in the relevant art that the inventor or a joint inventor, or for applications subject to pre-AIA 35 U.S.C. 112, the inventor(s), at the time the application was filed, had possession of the claimed invention.
Section 2163 of the M.P.E.P. states that the written description requirement for a claimed genus may be satisfied through sufficient description of a representative number of species by actual reduction to practice, or by disclosure of relevant, identifying characteristics, i.e., structure or other physical and/or chemical properties, by functional characteristics coupled with a known or disclosed correlation between function and structure, or by a combination of such identifying characteristics, sufficient to show the applicant was in possession of the claimed genus. A “representative number of species” means that the species which are adequately described are representative of the entire genus. See, e.g., AbbVie Deutschland GMBH v. Janssen Biotech, 111 USPQ2d 1780, 1790 (Fed. Cir. 2014). Thus, when there is substantial variation within the genus, one must describe a sufficient variety of species to reflect the variation within the genus to provide a “representative number” of species. The “structural features common to the members of the genus” needed for one of skill in the art to ‘visualize or recognize’ the members of the genus takes into account the state of the art at the time of the invention.
Claim 11 is reliant on a genus of immune cells characterized by expressing a receptor which is a chimeric antigen receptor defined, in part, by having the Vh portion comprising heavy chain CDRs of Table 3, or by having the Vl portion comprising light chain CDRs of Table 3a, or by having CDR sequences having up to 4 substitutions, insertions or deletions in each CDR. It is noted that the CDR sequences of Table 3 comprises sequences of 5 amino acids to 17 amino acids. Four substitutions out of 5 amino acids is equivalent to a variant having only 20% sequence identity; four deletions out of 5 amino acids leaves only a single amino acid; and four insertions at any location in a CDR having 5 amino acids would almost double the length of the CDR. The CDRs of Table 3 encompass SEQ ID NO:31-65 and 151-156. The specification fails to describe CDR sequences having any substitutions, insertions or deletion wherein the CDR sequences constitute the antibody paratope of the first receptor of the immune cell which is required to bind to EGFR. Claim 11 is also reliant in part on a genus of immune cells characterized by expressing a receptor which is a chimeric antigen receptor defined, in part, only by the heavy chain CDRs from Table 3; and as an alternative embodiment, a genus of immune cells characterized by expressing a receptor which is a chimeric antigen receptor defined, in part, only by the light chain CDRs from Table 3.
The specification fails to provide a written description of the genus of extracellular ligand binding domain of the second receptor, wherein the binding domain was characterized only by (i) the heavy chain CDRs of Table 3, (ii) the light chain CDRs of Table 3, or having CDR sequences having up to 4 substitutions, insertions or deletions in each CDR.
Claim 12 is reliant, in part, on a genus of immune cells characterized by expressing a receptor which is a chimeric antigen receptor defined, in part, by having the Vh portion comprising the Vh sequences pf Table 2, or as an alternative embodiment, by having the Vl portion comprising Vl sequences of Table 2, or by comprising sequences having at least 70% , 85%, 90% or 95% identity to the Vh or Vl of Table 2. Claim 13 is reliant on a genus of immune cells characterized by expressing a receptor which is a chimeric antigen receptor defined, in part, as a sequence having at least 85%, 90%, 95%, 97% or 99% identity to the sequences set forth in Table 1. Claim 14 is reliant on a genus of immune cells characterized by expressing a receptor which is a chimeric antigen receptor comprising a first receptor which is an scFv sequence having at least 85%, 90%, 95%, 97% or 99% identity to SEQ ID NO: 9-18.
Claim 16 is reliant on the immune cell of claim 1, wherein the LILRB1 intracellular domain comprises a sequence at least 90%, 95%, 97% or 99% identical to SEQ ID NO:129. Claim 18 is reliant on the immune cell of claim 1, wherein the LILRB1 transmembrane domain or a functional variant comprises a sequence at least 90%, 95%, 97% or 99% identical to SEQ ID NO:133. Claim 20 is reliant on the immune cell of claim 1, wherein the LILRB1 hinge domain comprises a sequence at least 90%, 95%, 97% or 99% identical to SEQ ID NO:125, 126, or 132. Claim 21 is reliant on the immune cell of claim 20, wherein the second receptor comprises a LILRB1 intracellular, transmembrane, and hinge domain, a functional variant of each of the intracellular, transmembrane or hinge domain, and combinations thereof, wherein the LILRB1 intracellular domain comprises a sequence having at least 95% identity to SEQ ID NO: 130 or 131, wherein the LILRB1 intracellular domain comprises the ITIMs of SEQ ID NO: 115, 116, 117 and 118.
It is well-known in the art that the CDR regions arranged in appropriate spatial orientation provide the antibody paratope which determines antigen recognition. In claim 11, variants comprising up to 4 substitutions, deletions and insertions into the CDR regions are claimed. Beyond the presence of the CDR regions in Table 3, the specification fails to provide any written description of alterations within these regions that would preserve the binding to the EGFR. Claim 11 relies on only on a genus of the heavy chain CDRS of Table 3 which can be paired with any light chain and thus, any light chain CDRs for binding to EGFR and as an alternative embodiment, relies only on the genus of light chain CDRs of Table 3, which can be paired with any heavy chain for binding to EGFR. The specification describes only
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This fails to describe a genus of immune cells, wherein the first receptor is chimeric antigen receptor, wherein the extracellular ligand binding domain of the first receptor is defined only by the heavy chain CDRs of Table 3, and the genus of immune cells wherein the first re receptor is chimeric antigen receptor, wherein the extracellular ligand binding domain of the first receptor is defined only by the light chain CDRs of Table 3 and the genus of immune cells having up to 4 substitutions, insertions or deletions in “each CDR”.
Claim 12 embodies the immune cell of claim 11 wherein the extracellular binding domain of the first receptor comprises a variable heavy portion having a sequence from the Vh sequences of Table 2, or as an alternative embodiment, the extracellular binding domain of the first receptor comprises a variable light portion having a sequence from the Vl sequences of Tale 2, or as an alternative embodiment, sequence having at least 70%, 85%,90% or 95% identity thereto. The specification describes:
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This fails to describe a genus of immune cells, wherein the first receptor is chimeric antigen receptor, wherein the extracellular ligand binding domain of the first receptor is defined only by the heavy chain Vh of Table 2, or the genus of immune cells wherein the first receptor is chimeric antigen receptor, wherein the extracellular ligand binding domain of the first receptor is defined only by the light chain Vl of Table 2 or the genus of immune cells wherein the first receptor is a chimeric antigen receptor, wherein the extracellular ligand binding domain of the first receptor has sequences having at least 70%, 85%, 90% or 95% identity to the Vh or Vl sequences of Table 2. It is noted that because claim 11 allows for alterations in the CDR sequences, claim 12 requiring sequences of at least 70%, 85%, 90% or95% identity does not insure that the CDR sequence from Table 3 will not be altered in the sequences of at least 70%, 85%, 90% or95% identity.
Claim 13 is reliant on genus of immune cells of claim 12 wherein the extracellular ligand binding domain of the first receptor comprises a sequence selected from the sequences of Table 1, or a sequence having at least 85%, 90% 95%, 97% or 99% identity thereto. Table 1 provides the sequences of the anti-EGFR scFv of SEQ ID NO: 9-18. This fails to provide an adequate written description of sequence having at least 85%, 90% 95%, 97% or 99% identity thereto. It is noted that claim 13 is dependent on claim 11, wherein up to 4 substitutions, deletions, insertion are permitted within each CDR. Thus, sequence having at least 85%, 90% 95%, 97% or 99% identity to SEQ ID NO: 9-18 does not insure that the CDR regions of Table 3 do not have substitutions, deletions, or insertions relative to Table 3 CDR sequences.
The specification has not described a representative number of variants for all of the variants encompassed by the claims that would serve to characterized the claimed genus. Neither has the specification correlated a minimum structure with a desired function for the extracellular ligand binding domain of the first receptor, or the LILRB1 intracellular domain, transmembrane domain or hinge domain. One of skill in the art would not be able to envisage an extracellular ligand binding domain of claims 11-14 other than an antibody or scFv comprising a heavy chain variable region from Table 2 paired with the light chain variable region taught by the specification to provide the antibody paratope for binding to EGFR and also indicated in Table 2, or the scFv sequences of SEQ ID NO: 9-18. One of skill in the art would not be able to envisage the LILRB1 intracellular, transmembrane or hinge domain other than (a)SEQ ID NO: 129, 130 or 131, or comprising the ITIMS of SEQ ID NO: 115-118 for the intracellular domain, (b)SEQ ID NO: 133 for the transmembrane domain, or (c)SEQ ID NO: 125, 126 or 132 for the hinge domain as one could with a well-described genus. One of skill in the art world reasonably conclude that applicant was not in possession of the invention at the time of filing.
Claims 31-47, 67-84 and 86-91 are included in this rejection because the claims are dependent on a claim addressed above.
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 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 1-4, 6-14, 67-74 86-88, 90 and 91 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Kamb et al (WO2021/030149).
Claims 1-4, 6-14, 67-74 86-88, 90 and 91 are rejected under 35 U.S.C. 102(a)(2) as being anticipated by Kamb et al (WO2021/030149).
Kamb et al disclose an immune cell, comprising: (1) a first engineered receptor, the first engineered receptor comprising a transmembrane region and an extracellular region, the extracellular region comprising a first ligand binding domain capable of specifically binding an EGFR ligand; and (2) a second engineered receptor, the second engineered receptor comprising a transmembrane region and an extracellular region, the extracellular region comprising a second ligand binding domain capable of specifically binding an HLA-A*02 allele, wherein binding of the first ligand binding domain to the EGFR ligand activates or promotes activation of the immune cell by the first receptor, and wherein binding of the second ligand binding domain to the HLA-A*02 allele inhibits activation of the immune cell by the first receptor (claim 87 of ‘149) which meets those limitation of instant claim 1, part (b) and claims4. Kamb et al disclose that the first ligand binding domain comprises SEQ ID NO: 391 (claim 88 of ‘149), which is identical to the instant SEQ ID NO: 18,
BIZ67783
ID BIZ67783 standard; protein; 245 AA.
XX
AC BIZ67783;
XX
DT 01-APR-2021 (first entry)
XX
DE Anti-EGFR single chain antibody (ScFv), SEQ 391.
XX
KW EGFR; cancer; cell culture; cell therapy; cytostatic; immunotherapy;
KW single chain antibody; therapeutic.
XX
OS Synthetic.
OS Unidentified.
XX
CC PN WO2021030149-A1.
XX
CC PD 18-FEB-2021.
XX
CC PF 06-AUG-2020; 2020WO-US045228.
XX
PR 09-AUG-2019; 2019US-0885093P.
PR 06-APR-2020; 2020US-0005670P.
XX
CC PA (ATWO-) A2 BIOTHERAPEUTICS INC.
XX
CC PI Kamb CA, Kamb A, Xu H;
XX
DR WPI; 2021-17756V/021.
XX
XX
CC PS Claim 28; SEQ ID NO 391; 210pp; English.
XX
XX
SQ Sequence 245 AA;
Query Match 100.0%; Score 1291; DB 33; Length 245;
Best Local Similarity 100.0%;
Matches 245; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGDYYWSWIRQPPGKGLEWIGYIYYSGSTD 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 QVQLQESGPGLVKPSQTLSLTCTVSGGSISSGDYYWSWIRQPPGKGLEWIGYIYYSGSTD 60
Qy 61 YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARVSIFGVGTFDYWGQGTLVTVS 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 YNPSLKSRVTMSVDTSKNQFSLKVNSVTAADTAVYYCARVSIFGVGTFDYWGQGTLVTVS 120
Qy 121 SGGGGSGGGGSGGGGSGGEIVMTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 SGGGGSGGGGSGGGGSGGEIVMTQSPATLSLSPGERATLSCRASQSVSSYLAWYQQKPGQ 180
Qy 181 APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCHQYGSTPLTFGGGT 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 APRLLIYDASNRATGIPARFSGSGSGTDFTLTISSLEPEDFAVYYCHQYGSTPLTFGGGT 240
Qy 241 KAEIK 245
|||||
Db 241 KAEIK 245
thus, meeting the limitations of instant claims 1(a), 6-10 and 11-14.
Kamb et al disclose that the inhibitory receptor comprises an scFv comprising SEQ ID NO: 57,
BIZ67449
ID BIZ67449 standard; protein; 247 AA.
XX
AC BIZ67449;
XX
DT 01-APR-2021 (first entry)
XX
DE Anti-HLA-A*02 single chain antibody, SEQ 57.
XX
KW HLA-A*02; cancer; cell culture; cell therapy; cytostatic; immunotherapy;
KW single chain antibody; therapeutic.
XX
OS Synthetic.
OS Unidentified.
XX
CC PN WO2021030149-A1.
XX
CC PD 18-FEB-2021.
XX
CC PF 06-AUG-2020; 2020WO-US045228.
XX
PR 09-AUG-2019; 2019US-0885093P.
PR 06-APR-2020; 2020US-0005670P.
XX
CC PA (ATWO-) A2 BIOTHERAPEUTICS INC.
XX
CC PI Kamb CA, Kamb A, Xu H;
XX
DR WPI; 2021-17756V/021.
XX
XX
CC PS Claim 39; SEQ ID NO 57; 210pp; English.
XX
XX
SQ Sequence 247 AA;
Query Match 100.0%; Score 1312; DB 33; Length 247;
Best Local Similarity 100.0%;
Matches 247; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYHIHWVRQAPGQGLEWIGWIYPGNVNTEY 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 QVQLVQSGAEVKKPGSSVKVSCKASGYTFTSYHIHWVRQAPGQGLEWIGWIYPGNVNTEY 60
Qy 61 NEKFKGKATITADESTNTAYMELSSLRSEDTAVYYCAREEITYAMDYWGQGTLVTVSSGG 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 NEKFKGKATITADESTNTAYMELSSLRSEDTAVYYCAREEITYAMDYWGQGTLVTVSSGG 120
Qy 121 GGSGGGGSGGGGSGGDIQMTQSPSTLSASVGDRVTITCRSSQSIVHSNGNTYLEWYQQKP 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GGSGGGGSGGGGSGGDIQMTQSPSTLSASVGDRVTITCRSSQSIVHSNGNTYLEWYQQKP 180
Qy 181 GKAPKLLIYKVSNRFSGVPARFSGSGSGTEFTLTISSLQPDDFATYYCFQGSHVPRTFGQ 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GKAPKLLIYKVSNRFSGVPARFSGSGSGTEFTLTISSLQPDDFATYYCFQGSHVPRTFGQ 240
Qy 241 GTKVEVK 247
|||||||
Db 241 GTKVEVK 247
which is identical to the instant SEQ ID NO: 93, thus meeting the limitations of instant claims 6-9.
Kamb et al disclose that the immune cells of the invention are T cells (claim 53 of ‘149 and paragraph [0063]) which meets the embodiments of instant claim 88. Kamb et al disclose that the inventive T cells include including autologous T cells and T cell lines (paragraph [0478]) which meet the limitations of claims 90 and 91, respectively. Kamb et al disclose that the activator CAR comprises a CD8 hinge (page 63, paragraph [0257]), a CD8 transmembrane region (page 64, paragraph [0262], line 5) and an intracellular domain derived from CD28, 4-1BB or CD3ζ (page 80, paragraph [0345], lines 7, 8 and 13) which meets the limitations of claim 10. Kamb et al disclose that the inhibitory CAR comprises a LILRB1 hinge domain, a LILRB1transmembrane domain and a LILRB1 intracellular domain (page 89, paragraph [0385]) which meets the limitations of instant claim 1(b).
Kamb et al disclose a method of treating a subject in need thereof, comprising administering to the subject a therapeutically effective amount of a composition comprising immune cells comprising the engineered receptors of the disclosure wherein the immune cells express both engineered receptors in the same cell (pages 110-111, paragraphs [0487]-[0488]) and that any cancer wherein a plurality of the cancer cells express the first, activator ligand and do not express the second, inhibitor ligand is envisaged as within the scope of the instant disclosure (paragraph [0489]), which meets the limitations of instant claims 67-70. Kamb et al disclose that the second ligand is selected from the group including a HLA class I allele, wherein expression of the second ligand has been lost in the target cell from loss of heterozygosity, such as loss of the HLA-A*02 allele (page 3, paragraph [0013]) which meets that embodiment in claims 1(b), 2-4 and 70-73.. Kamb et al disclose that the first ligand binding domain comprises a single chain Fv antibody fragment (scFv) wherein the first ligand is EGFR (paragraphs [0015]-[0016]) which meet the limitations of an EGFR+ cancer in claims 67-73. Kamb et al disclose that the condition treated by the methods described herein is metastasis of melanoma cells, prostate cancer cells, testicular cancer cells, breast cancer cells, brain cancer cells, pancreatic cancer cells, colon cancer cells, thyroid cancer cells, stomach cancer cells, lung cancer cells, ovarian cancer cells, Kaposi's sarcoma cells, skin cancer cells, renal cancer cells, head or neck cancer cells, throat cancer cells, squamous carcinoma cells, bladder cancer cells, osteosarcoma cells, cervical cancer cells, endometrial cancer cells, esophageal cancer cells, liver cancer cells, or kidney cancer cells (paragraph [0488]) which meets the limitations of claim 74 for treatment of metastases.
Kamb et al discloses kits and articles of manufacture comprising the immune cells comprising the engineered receptors wherein the kit comprises articles such as vials, syringes and instructions for use (paragraph [0504]) which meets the limitations of claims 86 and 87.
The applied reference has a common inventor with the instant application. Based upon the earlier effectively filed date of the reference, it constitutes prior art under 35 U.S.C. 102(a)(2). This rejection under 35 U.S.C. 102(a)(2) might be overcome by: (1) a showing under 37 CFR 1.130(a) that the subject matter disclosed in the reference was obtained directly or indirectly from the inventor or a joint inventor of this application and is thus not prior art in accordance with 35 U.S.C. 102(b)(2)(A); (2) a showing under 37 CFR 1.130(b) of a prior public disclosure under 35 U.S.C. 102(b)(2)(B) if the same invention is not being claimed; or (3) a statement pursuant to 35 U.S.C. 102(b)(2)(C) establishing that, not later than the effective filing date of the claimed invention, the subject matter disclosed in the reference and the claimed invention were either owned by the same person or subject to an obligation of assignment to the same person or subject to a joint research agreement.
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 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.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-4, 6-14, 67-81, 84, 86-88, 90 and 91 are rejected under 35 U.S.C. 103 as being unpatentable over Kamb et al (WO2021/030149) in view of Gross et al (WO2019/068007, cited in the prior Office action).
Kamb et al teach the limitations of claims 1-4, 6-14, 67-74, 86-88, 90 and 91 for the reasons set forth above. Kamb et al also teach that the treatment of cancer with the inventive T cells can result in a reduction in size of a tumor and that preferably, after treatment, tumor size is reduced by 5% (paragraph [0490]) which meets the limitations of claims 76 and 77. Kamb et al teach that the treatment of cancer with the inventive T cells can result reduction of tumor size of greater than 75% (paragraph [0490]). One of skill in the art would understand that a reduction of tumor size of greater than 75% includes reduction of tumor size by 100% and is commensurate with elimination of the tumor, thus fulfilling the limitations of claim 78. Kamb et al teach the treatment of cancer with the inventive T cells can result in in a decrease in tumor growth rate most preferably reduced by at least 75%. (paragraph [0498]). One of skill in the art would understand that a reduction of at least 75% includes a 100% reduction commensurate with arresting the growth of the tumor in the subject of instant claim 79. Kamb et al teach Kamb et al teach that cells expressing an EGFR CAR activator and an HLA-A*02 LIR-1 blocker are activated by EGFR+/HL A- A* 02- target cells but not EGFR+/HLA- A*02+ cells. (paragraph [0071]) and that hemizygous tumor cells which express only Target A and have lost Target B due to loss of heterozygosity (LOH), in a background of normal cells expressing both Target A and Target B. are amenable to the inventive cytotoxic therapeutics that are blocked by Target B and activated by Target A, thereby selectively killing tumors (paragraph [0041]) thus meeting the limitations of claim 81.
Kamb et al do not specifically teach that a) the genotype or expression level of a non-target antigen in non-malignant cells and cancer cells of the subject is determined; b) the expression level of EGFR in cancer cells of the subject is determined; © the subject is administered an effective amount of the immune cells of claim 88 if the non-malignant cells express the non-target antigen and the cancer cells express do not express the non-target antigen, and the cancer cells are EGFR positive.
Gross et al teach [0102] a method of providing a population of safe redirected immune effector cells for treating cancer in a patient having a tumor characterized by LOH, wherein the safe redirected immune cells are obtained by (i) identifying a single allelic variant of a polymorphic cell surface epitope that is not expressed by cells of the tumor due to LOH, but that is expressed by the cells of the normal tissue, (ii) identifying a non-polymorphic cell surface epitope of an antigen, wherein said epitope is a tumor- associated antigen or is shared by cells at least of related tumor and normal tissue in said cancer patient; (iii) selecting at least one nucleic acid molecule defining an iCAR as defined herein and at least one nucleic acid molecule comprising a nucleotide sequence encoding an aCAR as defined herein, wherein the iCAR comprises an extracellular domain that specifically binds to a cell surface epitope of (i) and the aCAR comprises an extracellular domain that specifically binds to a cell surface epitope of (ii); (iv) preparing at least one population of safe redirected effector immune cells by transfecting effector immune cells with the nucleic acid molecules of (iii) or transducing effector immune cells with the vectors of (iii).
It would have been prima facie obvious at the time prior to the effective filing date to identify a single allelic variant of an HLA allele that is not expressed by cells of the tumor due to LOH in the cancer cells of a subject; determine that the tumor- associated antigen, EGFR is expressed in the cancer cells of the patient; administer an effective amount of a T cell expressing an inhibitory CAR-T cell, wherein the iCAR comprises a scFv that binds to the cell surface allelic variant determined in part (i) and wherein the T cell also expresses an activating CAR which comprises a scFv that binds to EGFR to a subject if non-malignant cells of the subject express the allelic variant that is not expressed by cells of the tumor , and the cancer cells do not express the allelic variant and the cancer cells of the subject are EGFR positive.
One of skill in the art would have been motivated to do so in order to provide a personalized engineered T cell expressing a iCAR comprising an scFv that binds to a HLA specifically lost by LOH and an activating CAR comprising an scFv that binds to EGFR. One of skill in the art would have been motivated to do so because Kamb et al teach that cells expressing an EGFR CAR activator and an HLA-A*02 LIR-1 blocker are activated by EGFR+/HLA-A*02 negative target cells but not EGFR+/HLA-A*02 positive cells. One of skill in the art would understand that selecting the iCAR by the method of Gross et al would avoid “on-target” (EGFR), “off tumor” (normal cells expressing EGFR) toxicity, thus meeting the limitations of claim 84.
Claims 1-4, 6-14, 16, 18, 20, 21, 67-74, 86-88, 90 and 91 are rejected under 35 U.S.C. 103 as being unpatentable over Kamb et al (WO2021/030149) in view of Kamb et al (WO2021/119489, priority to 63/946,888)
Kamb et al (‘149) teach the limitations of claims 1-4, 6-14, 67-74, 86-88, 90 and 91 for the reasons set forth above. Kamb et al do not specifically teach that the LILRB1 intracellular, transmembrane and hinge domain together comprise the sequence of SEQ ID NO: 131.
Kamb et al (‘489) teach inhibitory chimeric antigen receptors having the hinge, transmembrane region, and/or intracellular domain of LILRB1 (paragraph [0005]). Kamb et al (‘489) teach that SEQ ID NO:2 and 3 having the LILBR1 hinge, transmembrane and intracellular domain. SEQ ID NO:2 is identical to the instant SEQ ID NO: 130 and SEQID NO:3 is identical to the instant SEQ ID NO: 131.
It would have been prima facie obvious at the time prior to the effective filing date to use SEQ ID NO:2 or 3 of Kamb et al (‘489) as the sequences of the LILRB1 hinge, transmembrane and intercellular domain of the iCAR of Kamb et al (‘149). One of skill in the art would have been motivated to do so because Kamb et al (‘149) teach that the inhibitor CAR had LILRB1 hinge, transmembrane and intercellular domain, and Kamb et al (‘489) teach the specific sequences for the domains. SEQ ID NO:2 and 3 of Kamb et al (‘489) meet the limitations of claim 16, 18 and 20 as well as 21 at least because SEQ ID NO: 129 of claim 16 is residues 67-233 of SEQ ID NO: 3, SEQ ID NO: 133 of claim 18 is residues 44-66 of SEQ ID NO: 3; and SEQ ID NO: 126 of claim 20 is residues 1-43 of SEQ ID NO: 3.
Claims 1-4, 6-14, 16, 18, 20, 21, 67-74, 86-88, 90 and 91 are rejected under 35 U.S.C. 103 as being unpatentable over Kamb et al (‘149) and Kamb et al (‘489) as applied to claims 1-4, 6-14, 16, 18, 20, 21, 67-74, 86-88, 90 and 91 above, and further in view of Ashton-Rickardt (WO2021/216978) as evidenced by Paddison et al (Genes & Development, 2002, Vol. 16, pp. 948-958).
The combined teachings of Kamb et al (‘149) and Kamb et al(‘489) render obvious the limitations of claims 1-4, 6-14, 16, 18, 20, 21, 67-74, 86-88, 90 and 91 for the reasons set forth above. Neither of Kamb et al nor Kamb et al teach the immune cell of claim 21, wherein expression of an MHC I gene has been reduced or eliminated, wherein the MHC I gene is B2M, wherein the immune cell comprises an iRNA against the B2M mRNA of SEQ ID NO: 172, wherein the iRNA is capable of inducing degradation of the B2M mRNA.
Ashton-Rickardt teaches SEQ ID NO: 10, which is the sequence of encoding human B2M RNA with adjacent sequences (page 7, line 17 to page 8, line 15). SEQ ID NO: 10 comprises the instant SEQ ID NO: 172::
GenCore version 6.5.2
Copyright (c) 1993 - 2026 Biocceleration Ltd.
OM nucleic - nucleic search, using sw model
Run on: June 10, 2026, 22:47:28 ; Search time 1 Seconds
(without alignments)
3.159 Million cell updates/sec
Title: US-18-165-623-172
Perfect score: 943
Sequence: 1 attcctgaagctgacagcat..........aaataaatcataaaacttga 943
Scoring table: IDENTITY_NUC
Gapop 10.0 , Gapext 1.0
Searched: 1 seqs, 1675 residues
Total number of hits satisfying chosen parameters: 2
Minimum DB seq length: 0
Maximum DB seq length: inf
Post-processing: Minimum Match 0%
Maximum Match 100%
Listing first 50 summaries
Database : NASEQ2_06102026_184725.fasta:*
SUMMARIES
%
Result Query
No. Score Match Length DB ID Description
----------------------------------------------------------------------------
1 943 100.0 1675 1 NASEQ2_06102026_184725
c 2 33.2 3.5 1675 1 NASEQ2_06102026_184725
ALIGNMENTS
RESULT 1
NASEQ2_06102026_184725
Query Match 100.0%; Score 943; DB 1; Length 1675;
Best Local Similarity 100.0%;
Matches 943; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 ATTCCTGAAGCTGACAGCATTCGGGCCGAGATGTCTCGCTCCGTGGCCTTAGCTGTGCTC 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 ATTCCTGAAGCTGACAGCATTCGGGCCGAGATGTCTCGCTCCGTGGCCTTAGCTGTGCTC 60
Qy 61 GCGCTACTCTCTCTTTCTGGCCTGGAGGCTATCCAGCGTACTCCAAAGATTCAGGTTTAC 120
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 GCGCTACTCTCTCTTTCTGGCCTGGAGGCTATCCAGCGTACTCCAAAGATTCAGGTTTAC 120
Qy 121 TCACGTCATCCAGCAGAGAATGGAAAGTCAAATTTCCTGAATTGCTATGTGTCTGGGTTT 180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 TCACGTCATCCAGCAGAGAATGGAAAGTCAAATTTCCTGAATTGCTATGTGTCTGGGTTT 180
Qy 181 CATCCATCCGACATTGAAGTTGACTTACTGAAGAATGGAGAGAGAATTGAAAAAGTGGAG 240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 CATCCATCCGACATTGAAGTTGACTTACTGAAGAATGGAGAGAGAATTGAAAAAGTGGAG 240
Qy 241 CATTCAGACTTGTCTTTCAGCAAGGACTGGTCTTTCTATCTCTTGTACTACACTGAATTC 300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 CATTCAGACTTGTCTTTCAGCAAGGACTGGTCTTTCTATCTCTTGTACTACACTGAATTC 300
Qy 301 ACCCCCACTGAAAAAGATGAGTATGCCTGCCGTGTGAACCATGTGACTTTGTCACAGCCC 360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 ACCCCCACTGAAAAAGATGAGTATGCCTGCCGTGTGAACCATGTGACTTTGTCACAGCCC 360
Qy 361 AAGATAGTTAAGTGGGATCGAGACATGTAAGCAGCATCATGGAGGTTTGAAGATGCCGCA 420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 AAGATAGTTAAGTGGGATCGAGACATGTAAGCAGCATCATGGAGGTTTGAAGATGCCGCA 420
Qy 421 TTTGGATTGGATGAATTCCAAATTCTGCTTGCTTGCTTTTTAATATTGATATGCTTATAC 480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 TTTGGATTGGATGAATTCCAAATTCTGCTTGCTTGCTTTTTAATATTGATATGCTTATAC 480
Qy 481 ACTTACACTTTATGCACAAAATGTAGGGTTATAATAATGTTAACATGGACATGATCTTCT 540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 ACTTACACTTTATGCACAAAATGTAGGGTTATAATAATGTTAACATGGACATGATCTTCT 540
Qy 541 TTATAATTCTACTTTGAGTGCTGTCTCCATGTTTGATGTATCTGAGCAGGTTGCTCCACA 600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 TTATAATTCTACTTTGAGTGCTGTCTCCATGTTTGATGTATCTGAGCAGGTTGCTCCACA 600
Qy 601 GGTAGCTCTAGGAGGGCTGGCAACTTAGAGGTGGGGAGCAGAGAATTCTCTTATCCAACA 660
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 GGTAGCTCTAGGAGGGCTGGCAACTTAGAGGTGGGGAGCAGAGAATTCTCTTATCCAACA 660
Qy 661 TCAACATCTTGGTCAGATTTGAACTCTTCAATCTCTTGCACTCAAAGCTTGTTAAGATAG 720
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 TCAACATCTTGGTCAGATTTGAACTCTTCAATCTCTTGCACTCAAAGCTTGTTAAGATAG 720
Qy 721 TTAAGCGTGCATAAGTTAACTTCCAATTTACATACTCTGCTTAGAATTTGGGGGAAAATT 780
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 TTAAGCGTGCATAAGTTAACTTCCAATTTACATACTCTGCTTAGAATTTGGGGGAAAATT 780
Qy 781 TAGAAATATAATTGACAGGATTATTGGAAATTTGTTATAATGAATGAAACATTTTGTCAT 840
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 TAGAAATATAATTGACAGGATTATTGGAAATTTGTTATAATGAATGAAACATTTTGTCAT 840
Qy 841 ATAAGATTCATATTTACTTCTTATACATTTGATAAAGTAAGGCATGGTTGTGGTTAATCT 900
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 ATAAGATTCATATTTACTTCTTATACATTTGATAAAGTAAGGCATGGTTGTGGTTAATCT 900
Qy 901 GGTTTATTTTTGTTCCACAAGTTAAATAAATCATAAAACTTGA 943
|||||||||||||||||||||||||||||||||||||||||||
Db 901 GGTTTATTTTTGTTCCACAAGTTAAATAAATCATAAAACTTGA 943
Ashton-Rickardt teaches shRNA target locations across the B2M gene (Figure 7 and page 9, lines 22-23) and B2M degrading siRNA comprising one or more of SEQ ID NO: 1-5 (page 9, lines 9-16) which meets the imitations of claims 31-34. Ashton-Rickardt teaches that the B2M-modifying RNA may include a small hairpin RNA (shRNA) which includes siRNA sequence(s) selected from the group consisting of SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3, SEQ ID NO: 4, and SEQ ID NO: 5 and one or more hairpin loop sequences selected from the group consisting of SEQ ID NO: 6 and SEQ ID NO: 7 (page 10, lines 1-4) which meet the limitations of claims 31-35. Ashton-Rickardt teaches that shRNA is an artificial molecule with a tight hairpin turn that can be used to silence target gene expression by RNA interference. Ashton-Rickardt teaches that shRNA can be incorporated into plasmid vectors and integrated into genomic DNA for a stable expression. Ashton-Rickardt teaches that the shRNA is processed in vivo to release the incorporated siRNA.
Paddison et al provide evidence that vectors comprising the shRNA are double stranded and have between 19-29 base homology with the target gene (page 955. Legend for Figure 4(A)), thus the shRNA of Ashton-Rickardt fulfills the requirement of claim 36 for a first sequence having a sequence complementary to the sequence of B2M mRNA and a second sequence having from 5’ to 3’ a sequence complementary to said first sequence.
Ashton-Rickardt teaches that B2M is a component of MHC I and is essential for cell surface MHC I peptide presentation, and stability of the peptide binding groove, such that absence of B2M results in very limited amounts of MHC I on the cell surface (page 7, lines 8-12). Ashton-Rickardt teaches that reduction or elimination of B2M expression by interference with B2M expression decreases recognition of the cell as foreign and thus reduces immunogenicity which allows for the use of non-autologous cells in transplants (page 7, lines 13-17). Ashton-Rickardt teaches the B2M-modifying sequences with additional chimeric antigen receptors and suicide genes for the production of targeted cells for cell-based immunotherapy (page 8, lines 30-33).
It would have been prima facie obvious at the time prior to the effective filing date to reduce or eliminate B2M expression in the immune cells rendered obvious by Kamb et al (‘149) and Kamb et al (‘489).. One of skill in the rt would have been motivated to do so to allow for the use of allogenic T cell lines to be used to make the immune cells expressing an inhibitory receptor comprising a scFv to a HLA lost by the cancer cells by LOH , and co-expressing an activating receptor comprising an scFv to a tumor associated antigen. Further, one of skill in the art would understand that the immune cell expression of the scFv to an HLA lost by heterozygosity could result in the binding of the scFv to the other immune cells in the composition of administered immune cells and thus limit effectiveness. Thus, by eliminating HLA expression by elimination or reduction of B2M would counter this possibility.
Claims 1-4, 6-14, 16, 18, 20, 21, 31, 32, 37-40, 67-74, 86-88, 90 and 91 are rejected under 35 U.S.C. 103 as being unpatentable over Kamb et al (‘149) and Kamb et al (‘489) as applied to claims 1-4, 6-14, 16, 18, 20, 21, 67-74, 86-88, 90 and 91 above, and further in view of Poirot et al (WO2015/136001).
The combined teachings of Kamb et al (‘149) and Kamb et al (‘489) render obvious the limitations of claims 1-4, 6-14, 16, 18, 20, 21, 67-74, 86-88, 90 and 91 for the reasons set forth above. Neither of Kamb et al nor Kamb et al teach the immune cell of claim 21, wherein expression or function of an MHC I gene has been reduced or eliminations, wherein the MHC I gene is B2M, wherein the immune cell comprises modified B2M, wherein the modification comprises one or more inactivating mutations of the endogenous gene encoding B2M, wherein the one or more inactivating mutations are introduced with a nucleic acid guided endonuclease in a complex with a guide nucleic acid that specifically targets SEQ ID NO: 170.
Poirot et al teach an engineered T cell, wherein said T cell expresses a rare cutting endonuclease able to selectively inactivate the gene encoding B2M by DNA cleavage, wherein said T cell comprises a nucleic acid encoding said rare-cutting endonuclease, wherein the rare cutting endonuclease is a RNA-guided endonuclease, and wherein said cleavage is the result of an exogenous nucleic acid that inhibit the expression of B2M (claims 24 , 26, 27, 29, 30 and 33 of ‘001) thus fulfilling the limitations of instant claims 31, 32, and 38-40 for inactivation by cleavage which is a “deletion”.. Poirot et al teach that the inventio provides improvement to immunotherapy because a allogeneic T cell engineered by the inventive method can be used to treat a subject as opposed to engineering of an autologous T cell (page 4, lines 16-30). Poirot et al teach that the engineered cell can further comprise a CAR directed against at least one antigen expressed at the surface of a malignant cell (claim 42 of ‘001).
SEQ ID NO: 2 od Poirot et al comprises instant SEQ ID NO: 170:
GenCore version 6.5.2
Copyright (c) 1993 - 2026 Biocceleration Ltd.
OM nucleic - nucleic search, using sw model
Run on: June 11, 2026, 19:31:59 ; Search time 1 Seconds
(without alignments)
88.804 Million cell updates/sec
Title: NASEQ1_06112026_153120
Perfect score: 6673
Sequence: 1 aatataagtggaggcgtcgc..........acttgatgtgttatctctta 6673
Scoring table: IDENTITY_NUC
Gapop 10.0 , Gapext 1.0
Searched: 1 seqs, 6654 residues
Total number of hits satisfying chosen parameters: 2
Minimum DB seq length: 0
Maximum DB seq length: inf
Post-processing: Minimum Match 0%
Maximum Match 100%
Listing first 50 summaries
Database : US-18-165-623-170.fasta:*
SUMMARIES
%
Result Query
No. Score Match Length DB ID Description
----------------------------------------------------------------------------
1 6654 99.7 6654 1 US-18-165-623-170 COMPOSITIONS AND M
c 2 34.2 0.5 6654 1 US-18-165-623-170 COMPOSITIONS AND M
ALIGNMENTS
RESULT 1
US-18-165-623-170
Query Match 99.7%; Score 6654; DB 1; Length 6654;
Best Local Similarity 100.0%;
Matches 6654; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 6 AAGTGGAGGCGTCGCGCTGGCGGGCATTCCTGAAGCTGACAGCATTCGGGCCGAGATGTC 65
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 AAGTGGAGGCGTCGCGCTGGCGGGCATTCCTGAAGCTGACAGCATTCGGGCCGAGATGTC 60
Qy 66 TCGCTCCGTGGCCTTAGCTGTGCTCGCGCTACTCTCTCTTTCTGGCCTGGAGGCTATCCA 125
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 TCGCTCCGTGGCCTTAGCTGTGCTCGCGCTACTCTCTCTTTCTGGCCTGGAGGCTATCCA 120
Qy 126 GCGTGAGTCTCTCCTACCCTCCCGCTCTGGTCCTTCCTCTCCCGCTCTGCACCCTCTGTG 185
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 GCGTGAGTCTCTCCTACCCTCCCGCTCTGGTCCTTCCTCTCCCGCTCTGCACCCTCTGTG 180
Qy 186 GCCCTCGCTGTGCTCTCTCGCTCCGTGACTTCCCTTCTCCAAGTTCTCCTTGGTGGCCCG 245
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 GCCCTCGCTGTGCTCTCTCGCTCCGTGACTTCCCTTCTCCAAGTTCTCCTTGGTGGCCCG 240
Qy 246 CCGTGGGGCTAGTCCAGGGCTGGATCTCGGGGAAGCGGCGGGGTGGCCTGGGAGTGGGGA 305
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 CCGTGGGGCTAGTCCAGGGCTGGATCTCGGGGAAGCGGCGGGGTGGCCTGGGAGTGGGGA 300
Qy 306 AGGGGGTGCGCACCCGGGACGCGCGCTACTTGCCCCTTTCGGCGGGGAGCAGGGGAGACC 365
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 AGGGGGTGCGCACCCGGGACGCGCGCTACTTGCCCCTTTCGGCGGGGAGCAGGGGAGACC 360
Qy 366 TTTGGCCTACGGCGACGGGAGGGTCGGGACAAAGTTTAGGGCGTCGATAAGCGTCAGAGC 425
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 TTTGGCCTACGGCGACGGGAGGGTCGGGACAAAGTTTAGGGCGTCGATAAGCGTCAGAGC 420
Qy 426 GCCGAGGTTGGGGGAGGGTTTCTCTTCCGCTCTTTCGCGGGGCCTCTGGCTCCCCCAGCG 485
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 GCCGAGGTTGGGGGAGGGTTTCTCTTCCGCTCTTTCGCGGGGCCTCTGGCTCCCCCAGCG 480
Qy 486 CAGCTGGAGTGGGGGACGGGTAGGCTCGTCCCAAAGGCGCGGCGCTGAGGTTTGTGAACG 545
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 CAGCTGGAGTGGGGGACGGGTAGGCTCGTCCCAAAGGCGCGGCGCTGAGGTTTGTGAACG 540
Qy 546 CGTGGAGGGGCGCTTGGGGTCTGGGGGAGGCGTCGCCCGGGTAAGCCTGTCTGCTGCGGC 605
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 CGTGGAGGGGCGCTTGGGGTCTGGGGGAGGCGTCGCCCGGGTAAGCCTGTCTGCTGCGGC 600
Qy 606 TCTGCTTCCCTTAGACTGGAGAGCTGTGGACTTCGTCTAGGCGCCCGCTAAGTTCGCATG 665
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 TCTGCTTCCCTTAGACTGGAGAGCTGTGGACTTCGTCTAGGCGCCCGCTAAGTTCGCATG 660
Qy 666 TCCTAGCACCTCTGGGTCTATGTGGGGCCACACCGTGGGGAGGAAACAGCACGCGACGTT 725
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 661 TCCTAGCACCTCTGGGTCTATGTGGGGCCACACCGTGGGGAGGAAACAGCACGCGACGTT 720
Qy 726 TGTAGAATGCTTGGCTGTGATACAAAGCGGTTTCGAATAATTAACTTATTTGTTCCCATC 785
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 721 TGTAGAATGCTTGGCTGTGATACAAAGCGGTTTCGAATAATTAACTTATTTGTTCCCATC 780
Qy 786 ACATGTCACTTTTAAAAAATTATAAGAACTACCCGTTATTGACATCTTTCTGTGTGCCAA 845
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 781 ACATGTCACTTTTAAAAAATTATAAGAACTACCCGTTATTGACATCTTTCTGTGTGCCAA 840
Qy 846 GGACTTTATGTGCTTTGCGTCATTTAATTTTGAAAACAGTTATCTTCCGCCATAGATAAC 905
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 841 GGACTTTATGTGCTTTGCGTCATTTAATTTTGAAAACAGTTATCTTCCGCCATAGATAAC 900
Qy 906 TACTATGGTTATCTTCTGCCTCTCACAGATGAAGAAACTAAGGCACCGAGATTTTAAGAA 965
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 901 TACTATGGTTATCTTCTGCCTCTCACAGATGAAGAAACTAAGGCACCGAGATTTTAAGAA 960
Qy 966 ACTTAATTACACAGGGGATAAATGGCAGCAATCGAGATTGAAGTCAAGCCTAACCAGGGC 1025
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 961 ACTTAATTACACAGGGGATAAATGGCAGCAATCGAGATTGAAGTCAAGCCTAACCAGGGC 1020
Qy 1026 TTTTGCGGGAGCGCATGCCTTTTGGCTGTAATTCGTGCATTTTTTTTTAAGAAAAACGCC 1085
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1021 TTTTGCGGGAGCGCATGCCTTTTGGCTGTAATTCGTGCATTTTTTTTTAAGAAAAACGCC 1080
Qy 1086 TGCCTTCTGCGTGAGATTCTCCAGAGCAAACTGGGCGGCATGGGCCCTGTGGTCTTTTCG 1145
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1081 TGCCTTCTGCGTGAGATTCTCCAGAGCAAACTGGGCGGCATGGGCCCTGTGGTCTTTTCG 1140
Qy 1146 TACAGAGGGCTTCCTCTTTGGCTCTTTGCCTGGTTGTTTCCAAGATGTACTGTGCCTCTT 1205
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1141 TACAGAGGGCTTCCTCTTTGGCTCTTTGCCTGGTTGTTTCCAAGATGTACTGTGCCTCTT 1200
Qy 1206 ACTTTCGGTTTTGAAAACATGAGGGGGTTGGGCGTGGTAGCTTACGCCTGTAATCCCAGC 1265
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1201 ACTTTCGGTTTTGAAAACATGAGGGGGTTGGGCGTGGTAGCTTACGCCTGTAATCCCAGC 1260
Qy 1266 ACTTAGGGAGGCCGAGGCGGGAGGATGGCTTGAGGTCCGTAGTTGAGACCAGCCTGGCCA 1325
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1261 ACTTAGGGAGGCCGAGGCGGGAGGATGGCTTGAGGTCCGTAGTTGAGACCAGCCTGGCCA 1320
Qy 1326 ACATGGTGAAGCCTGGTCTCTACAAAAAATAATAACAAAAATTAGCCGGGTGTGGTGGCT 1385
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1321 ACATGGTGAAGCCTGGTCTCTACAAAAAATAATAACAAAAATTAGCCGGGTGTGGTGGCT 1380
Qy 1386 CGTGCCTGTGGTCCCAGCTGCTCCGGTGGCTGAGGCGGGAGGATCTCTTGAGCTTAGGCT 1445
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1381 CGTGCCTGTGGTCCCAGCTGCTCCGGTGGCTGAGGCGGGAGGATCTCTTGAGCTTAGGCT 1440
Qy 1446 TTTGAGCTATCATGGCGCCAGTGCACTCCAGCGTGGGCAACAGAGCGAGACCCTGTCTCT 1505
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1441 TTTGAGCTATCATGGCGCCAGTGCACTCCAGCGTGGGCAACAGAGCGAGACCCTGTCTCT 1500
Qy 1506 CAAAAAAGAAAAAAAAAAAAAAAGAAAGAGAAAAGAAAAGAAAGAAAGAAGTGAAGGTTT 1565
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1501 CAAAAAAGAAAAAAAAAAAAAAAGAAAGAGAAAAGAAAAGAAAGAAAGAAGTGAAGGTTT 1560
Qy 1566 GTCAGTCAGGGGAGCTGTAAAACCATTAATAAAGATAATCCAAGATGGTTACCAAGACTG 1625
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1561 GTCAGTCAGGGGAGCTGTAAAACCATTAATAAAGATAATCCAAGATGGTTACCAAGACTG 1620
Qy 1626 TTGAGGACGCCAGAGATCTTGAGCACTTTCTAAGTACCTGGCAATACACTAAGCGCGCTC 1685
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1621 TTGAGGACGCCAGAGATCTTGAGCACTTTCTAAGTACCTGGCAATACACTAAGCGCGCTC 1680
Qy 1686 ACCTTTTCCTCTGGCAAAACATGATCGAAAGCAGAATGTTTTGATCATGAGAAAATTGCA 1745
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1681 ACCTTTTCCTCTGGCAAAACATGATCGAAAGCAGAATGTTTTGATCATGAGAAAATTGCA 1740
Qy 1746 TTTAATTTGAATACAATTTATTTACAACATAAAGGATAATGTATATATCACCACCATTAC 1805
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1741 TTTAATTTGAATACAATTTATTTACAACATAAAGGATAATGTATATATCACCACCATTAC 1800
Qy 1806 TGGTATTTGCTGGTTATGTTAGATGTCATTTTAAAAAATAACAATCTGATATTTAAAAAA 1865
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1801 TGGTATTTGCTGGTTATGTTAGATGTCATTTTAAAAAATAACAATCTGATATTTAAAAAA 1860
Qy 1866 AAATCTTATTTTGAAAATTTCCAAAGTAATACATGCCATGCATAGACCATTTCTGGAAGA 1925
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1861 AAATCTTATTTTGAAAATTTCCAAAGTAATACATGCCATGCATAGACCATTTCTGGAAGA 1920
Qy 1926 TACCACAAGAAACATGTAATGATGATTGCCTCTGAAGGTCTATTTTCCTCCTCTGACCTG 1985
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1921 TACCACAAGAAACATGTAATGATGATTGCCTCTGAAGGTCTATTTTCCTCCTCTGACCTG 1980
Qy 1986 TGTGTGGGTTTTGTTTTTGTTTTACTGTGGGCATAAATTAATTTTTCAGTTAAGTTTTGG 2045
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1981 TGTGTGGGTTTTGTTTTTGTTTTACTGTGGGCATAAATTAATTTTTCAGTTAAGTTTTGG 2040
Qy 2046 AAGCTTAAATAACTCTCCAAAAGTCATAAAGCCAGTAACTGGTTGAGCCCAAATTCAAAC 2105
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2041 AAGCTTAAATAACTCTCCAAAAGTCATAAAGCCAGTAACTGGTTGAGCCCAAATTCAAAC 2100
Qy 2106 CCAGCCTGTCTGATACTTGTCCTCTTCTTAGAAAAGATTACAGTGATGCTCTCACAAAAT 2165
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2101 CCAGCCTGTCTGATACTTGTCCTCTTCTTAGAAAAGATTACAGTGATGCTCTCACAAAAT 2160
Qy 2166 CTTGCCGCCTTCCCTCAAACAGAGAGTTCCAGGCAGGATGAATCTGTGCTCTGATCCCTG 2225
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2161 CTTGCCGCCTTCCCTCAAACAGAGAGTTCCAGGCAGGATGAATCTGTGCTCTGATCCCTG 2220
Qy 2226 AGGCATTTAATATGTTCTTATTATTAGAAGCTCAGATGCAAAGAGCTCTCTTAGCTTTTA 2285
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2221 AGGCATTTAATATGTTCTTATTATTAGAAGCTCAGATGCAAAGAGCTCTCTTAGCTTTTA 2280
Qy 2286 ATGTTATGAAAAAAATCAGGTCTTCATTAGATTCCCCAATCCACCTCTTGATGGGGCTAG 2345
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2281 ATGTTATGAAAAAAATCAGGTCTTCATTAGATTCCCCAATCCACCTCTTGATGGGGCTAG 2340
Qy 2346 TAGCCTTTCCTTAATGATAGGGTGTTTCTAGAGAGATATATCTGGTCAAGGTGGCCTGGT 2405
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2341 TAGCCTTTCCTTAATGATAGGGTGTTTCTAGAGAGATATATCTGGTCAAGGTGGCCTGGT 2400
Qy 2406 ACTCCTCCTTCTCCCCACAGCCTCCCAGACAAGGAGGAGTAGCTGCCTTTTAGTGATCAT 2465
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2401 ACTCCTCCTTCTCCCCACAGCCTCCCAGACAAGGAGGAGTAGCTGCCTTTTAGTGATCAT 2460
Qy 2466 GTACCCTGAATATAAGTGTATTTAAAAGAATTTTATACACATATATTTAGTGTCAATCTG 2525
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2461 GTACCCTGAATATAAGTGTATTTAAAAGAATTTTATACACATATATTTAGTGTCAATCTG 2520
Qy 2526 TATATTTAGTAGCACTAACACTTCTCTTCATTTTCAATGAAAAATATAGAGTTTATAATA 2585
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2521 TATATTTAGTAGCACTAACACTTCTCTTCATTTTCAATGAAAAATATAGAGTTTATAATA 2580
Qy 2586 TTTTCTTCCCACTTCCCCATGGATGGTCTAGTCATGCCTCTCATTTTGGAAAGTACTGTT 2645
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2581 TTTTCTTCCCACTTCCCCATGGATGGTCTAGTCATGCCTCTCATTTTGGAAAGTACTGTT 2640
Qy 2646 TCTGAAACATTAGGCAATATATTCCCAACCTGGCTAGTTTACAGCAATCACCTGTGGATG 2705
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2641 TCTGAAACATTAGGCAATATATTCCCAACCTGGCTAGTTTACAGCAATCACCTGTGGATG 2700
Qy 2706 CTAATTAAAACGCAAATCCCACTGTCACATGCATTACTCCATTTGATCATAATGGAAAGT 2765
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2701 CTAATTAAAACGCAAATCCCACTGTCACATGCATTACTCCATTTGATCATAATGGAAAGT 2760
Qy 2766 ATGTTCTGTCCCATTTGCCATAGTCCTCACCTATCCCTGTTGTATTTTATCGGGTCCAAC 2825
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2761 ATGTTCTGTCCCATTTGCCATAGTCCTCACCTATCCCTGTTGTATTTTATCGGGTCCAAC 2820
Qy 2826 TCAACCATTTAAGGTATTTGCCAGCTCTTGTATGCATTTAGGTTTTGTTTCTTTGTTTTT 2885
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2821 TCAACCATTTAAGGTATTTGCCAGCTCTTGTATGCATTTAGGTTTTGTTTCTTTGTTTTT 2880
Qy 2886 TAGCTCATGAAATTAGGTACAAAGTCAGAGAGGGGTCTGGCATATAAAACCTCAGCAGAA 2945
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2881 TAGCTCATGAAATTAGGTACAAAGTCAGAGAGGGGTCTGGCATATAAAACCTCAGCAGAA 2940
Qy 2946 ATAAAGAGGTTTTGTTGTTTGGTAAGAACATACCTTGGGTTGGTTGGGCACGGTGGCTCG 3005
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 2941 ATAAAGAGGTTTTGTTGTTTGGTAAGAACATACCTTGGGTTGGTTGGGCACGGTGGCTCG 3000
Qy 3006 TGCCTGTAATCCCAACACTTTGGGAGGCCAAGGCAGGCTGATCACTTGAAGTTGGGAGTT 3065
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3001 TGCCTGTAATCCCAACACTTTGGGAGGCCAAGGCAGGCTGATCACTTGAAGTTGGGAGTT 3060
Qy 3066 CAAGACCAGCCTGGCCAACATGGTGAAATCCCGTCTCTACTGAAAATACAAAAATTAACC 3125
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3061 CAAGACCAGCCTGGCCAACATGGTGAAATCCCGTCTCTACTGAAAATACAAAAATTAACC 3120
Qy 3126 AGGCATGGTGGTGTGTGCCTGTAGTCCCAGGAATCACTTGAACCCAGGAGGCGGAGGTTG 3185
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3121 AGGCATGGTGGTGTGTGCCTGTAGTCCCAGGAATCACTTGAACCCAGGAGGCGGAGGTTG 3180
Qy 3186 CAGTGAGCTGAGATCTCACCACTGCACACTGCACTCCAGCCTGGGCAATGGAATGAGATT 3245
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3181 CAGTGAGCTGAGATCTCACCACTGCACACTGCACTCCAGCCTGGGCAATGGAATGAGATT 3240
Qy 3246 CCATCCCAAAAAATAAAAAAATAAAAAAATAAAGAACATACCTTGGGTTGATCCACTTAG 3305
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3241 CCATCCCAAAAAATAAAAAAATAAAAAAATAAAGAACATACCTTGGGTTGATCCACTTAG 3300
Qy 3306 GAACCTCAGATAATAACATCTGCCACGTATAGAGCAATTGCTATGTCCCAGGCACTCTAC 3365
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3301 GAACCTCAGATAATAACATCTGCCACGTATAGAGCAATTGCTATGTCCCAGGCACTCTAC 3360
Qy 3366 TAGACACTTCATACAGTTTAGAAAATCAGATGGGTGTAGATCAAGGCAGGAGCAGGAACC 3425
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3361 TAGACACTTCATACAGTTTAGAAAATCAGATGGGTGTAGATCAAGGCAGGAGCAGGAACC 3420
Qy 3426 AAAAAGAAAGGCATAAACATAAGAAAAAAAATGGAAGGGGTGGAAACAGAGTACAATAAC 3485
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3421 AAAAAGAAAGGCATAAACATAAGAAAAAAAATGGAAGGGGTGGAAACAGAGTACAATAAC 3480
Qy 3486 ATGAGTAATTTGATGGGGGCTATTATGAACTGAGAAATGAACTTTGAAAAGTATCTTGGG 3545
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3481 ATGAGTAATTTGATGGGGGCTATTATGAACTGAGAAATGAACTTTGAAAAGTATCTTGGG 3540
Qy 3546 GCCAAATCATGTAGACTCTTGAGTGATGTGTTAAGGAATGCTATGAGTGCTGAGAGGGCA 3605
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3541 GCCAAATCATGTAGACTCTTGAGTGATGTGTTAAGGAATGCTATGAGTGCTGAGAGGGCA 3600
Qy 3606 TCAGAAGTCCTTGAGAGCCTCCAGAGAAAGGCTCTTAAAAATGCAGCGCAATCTCCAGTG 3665
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3601 TCAGAAGTCCTTGAGAGCCTCCAGAGAAAGGCTCTTAAAAATGCAGCGCAATCTCCAGTG 3660
Qy 3666 ACAGAAGATACTGCTAGAAATCTGCTAGAAAAAAAACAAAAAAGGCATGTATAGAGGAAT 3725
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3661 ACAGAAGATACTGCTAGAAATCTGCTAGAAAAAAAACAAAAAAGGCATGTATAGAGGAAT 3720
Qy 3726 TATGAGGGAAAGATACCAAGTCACGGTTTATTCTTCAAAATGGAGGTGGCTTGTTGGGAA 3785
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3721 TATGAGGGAAAGATACCAAGTCACGGTTTATTCTTCAAAATGGAGGTGGCTTGTTGGGAA 3780
Qy 3786 GGTGGAAGCTCATTTGGCCAGAGTGGAAATGGAATTGGGAGAAATCGATGACCAAATGTA 3845
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3781 GGTGGAAGCTCATTTGGCCAGAGTGGAAATGGAATTGGGAGAAATCGATGACCAAATGTA 3840
Qy 3846 AACACTTGGTGCCTGATATAGCTTGACACCAAGTTAGCCCCAAGTGAAATACCCTGGCAA 3905
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3841 AACACTTGGTGCCTGATATAGCTTGACACCAAGTTAGCCCCAAGTGAAATACCCTGGCAA 3900
Qy 3906 TATTAATGTGTCTTTTCCCGATATTCCTCAGGTACTCCAAAGATTCAGGTTTACTCACGT 3965
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3901 TATTAATGTGTCTTTTCCCGATATTCCTCAGGTACTCCAAAGATTCAGGTTTACTCACGT 3960
Qy 3966 CATCCAGCAGAGAATGGAAAGTCAAATTTCCTGAATTGCTATGTGTCTGGGTTTCATCCA 4025
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 3961 CATCCAGCAGAGAATGGAAAGTCAAATTTCCTGAATTGCTATGTGTCTGGGTTTCATCCA 4020
Qy 4026 TCCGACATTGAAGTTGACTTACTGAAGAATGGAGAGAGAATTGAAAAAGTGGAGCATTCA 4085
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4021 TCCGACATTGAAGTTGACTTACTGAAGAATGGAGAGAGAATTGAAAAAGTGGAGCATTCA 4080
Qy 4086 GACTTGTCTTTCAGCAAGGACTGGTCTTTCTATCTCTTGTACTACACTGAATTCACCCCC 4145
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4081 GACTTGTCTTTCAGCAAGGACTGGTCTTTCTATCTCTTGTACTACACTGAATTCACCCCC 4140
Qy 4146 ACTGAAAAAGATGAGTATGCCTGCCGTGTGAACCATGTGACTTTGTCACAGCCCAAGATA 4205
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4141 ACTGAAAAAGATGAGTATGCCTGCCGTGTGAACCATGTGACTTTGTCACAGCCCAAGATA 4200
Qy 4206 GTTAAGTGGGGTAAGTCTTACATTCTTTTGTAAGCTGCTGAAAGTTGTGTATGAGTAGTC 4265
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4201 GTTAAGTGGGGTAAGTCTTACATTCTTTTGTAAGCTGCTGAAAGTTGTGTATGAGTAGTC 4260
Qy 4266 ATATCATAAAGCTGCTTTGATATAAAAAAGGTCTATGGCCATACTACCCTGAATGAGTCC 4325
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4261 ATATCATAAAGCTGCTTTGATATAAAAAAGGTCTATGGCCATACTACCCTGAATGAGTCC 4320
Qy 4326 CATCCCATCTGATATAAACAATCTGCATATTGGGATTGTCAGGGAATGTTCTTAAAGATC 4385
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4321 CATCCCATCTGATATAAACAATCTGCATATTGGGATTGTCAGGGAATGTTCTTAAAGATC 4380
Qy 4386 AGATTAGTGGCACCTGCTGAGATACTGATGCACAGCATGGTTTCTGAACCAGTAGTTTCC 4445
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4381 AGATTAGTGGCACCTGCTGAGATACTGATGCACAGCATGGTTTCTGAACCAGTAGTTTCC 4440
Qy 4446 CTGCAGTTGAGCAGGGAGCAGCAGCAGCACTTGCACAAATACATATACACTCTTAACACT 4505
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4441 CTGCAGTTGAGCAGGGAGCAGCAGCAGCACTTGCACAAATACATATACACTCTTAACACT 4500
Qy 4506 TCTTACCTACTGGCTTCCTCTAGCTTTTGTGGCAGCTTCAGGTATATTTAGCACTGAACG 4565
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4501 TCTTACCTACTGGCTTCCTCTAGCTTTTGTGGCAGCTTCAGGTATATTTAGCACTGAACG 4560
Qy 4566 AACATCTCAAGAAGGTATAGGCCTTTGTTTGTAAGTCCTGCTGTCCTAGCATCCTATAAT 4625
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4561 AACATCTCAAGAAGGTATAGGCCTTTGTTTGTAAGTCCTGCTGTCCTAGCATCCTATAAT 4620
Qy 4626 CCTGGACTTCTCCAGTACTTTCTGGCTGGATTGGTATCTGAGGCTAGTAGGAAGGGCTTG 4685
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4621 CCTGGACTTCTCCAGTACTTTCTGGCTGGATTGGTATCTGAGGCTAGTAGGAAGGGCTTG 4680
Qy 4686 TTCCTGCTGGGTAGCTCTAAACAATGTATTCATGGGTAGGAACAGCAGCCTATTCTGCCA 4745
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4681 TTCCTGCTGGGTAGCTCTAAACAATGTATTCATGGGTAGGAACAGCAGCCTATTCTGCCA 4740
Qy 4746 GCCTTATTTCTAACCATTTTAGACATTTGTTAGTACATGGTATTTTAAAAGTAAAACTTA 4805
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4741 GCCTTATTTCTAACCATTTTAGACATTTGTTAGTACATGGTATTTTAAAAGTAAAACTTA 4800
Qy 4806 ATGTCTTCCTTTTTTTTCTCCACTGTCTTTTTCATAGATCGAGACATGTAAGCAGCATCA 4865
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4801 ATGTCTTCCTTTTTTTTCTCCACTGTCTTTTTCATAGATCGAGACATGTAAGCAGCATCA 4860
Qy 4866 TGGAGGTAAGTTTTTGACCTTGAGAAAATGTTTTTGTTTCACTGTCCTGAGGACTATTTA 4925
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4861 TGGAGGTAAGTTTTTGACCTTGAGAAAATGTTTTTGTTTCACTGTCCTGAGGACTATTTA 4920
Qy 4926 TAGACAGCTCTAACATGATAACCCTCACTATGTGGAGAACATTGACAGAGTAACATTTTA 4985
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4921 TAGACAGCTCTAACATGATAACCCTCACTATGTGGAGAACATTGACAGAGTAACATTTTA 4980
Qy 4986 GCAGGGAAAGAAGAATCCTACAGGGTCATGTTCCCTTCTCCTGTGGAGTGGCATGAAGAA 5045
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 4981 GCAGGGAAAGAAGAATCCTACAGGGTCATGTTCCCTTCTCCTGTGGAGTGGCATGAAGAA 5040
Qy 5046 GGTGTATGGCCCCAGGTATGGCCATATTACTGACCCTCTACAGAGAGGGCAAAGGAACTG 5105
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5041 GGTGTATGGCCCCAGGTATGGCCATATTACTGACCCTCTACAGAGAGGGCAAAGGAACTG 5100
Qy 5106 CCAGTATGGTATTGCAGGATAAAGGCAGGTGGTTACCCACATTACCTGCAAGGCTTTGAT 5165
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5101 CCAGTATGGTATTGCAGGATAAAGGCAGGTGGTTACCCACATTACCTGCAAGGCTTTGAT 5160
Qy 5166 CTTTCTTCTGCCATTTCCACATTGGACATCTCTGCTGAGGAGAGAAAATGAACCACTCTT 5225
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5161 CTTTCTTCTGCCATTTCCACATTGGACATCTCTGCTGAGGAGAGAAAATGAACCACTCTT 5220
Qy 5226 TTCCTTTGTATAATGTTGTTTTATTCTTCAGACAGAAGAGAGGAGTTATACAGCTCTGCA 5285
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5221 TTCCTTTGTATAATGTTGTTTTATTCTTCAGACAGAAGAGAGGAGTTATACAGCTCTGCA 5280
Qy 5286 GACATCCCATTCCTGTATGGGGACTGTGTTTGCCTCTTAGAGGTTCCCAGGCCACTAGAG 5345
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5281 GACATCCCATTCCTGTATGGGGACTGTGTTTGCCTCTTAGAGGTTCCCAGGCCACTAGAG 5340
Qy 5346 GAGATAAAGGGAAACAGATTGTTATAACTTGATATAATGATACTATAATAGATGTAACTA 5405
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5341 GAGATAAAGGGAAACAGATTGTTATAACTTGATATAATGATACTATAATAGATGTAACTA 5400
Qy 5406 CAAGGAGCTCCAGAAGCAAGAGAGAGGGAGGAACTTGGACTTCTCTGCATCTTTAGTTGG 5465
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5401 CAAGGAGCTCCAGAAGCAAGAGAGAGGGAGGAACTTGGACTTCTCTGCATCTTTAGTTGG 5460
Qy 5466 AGTCCAAAGGCTTTTCAATGAAATTCTACTGCCCAGGGTACATTGATGCTGAAACCCCAT 5525
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5461 AGTCCAAAGGCTTTTCAATGAAATTCTACTGCCCAGGGTACATTGATGCTGAAACCCCAT 5520
Qy 5526 TCAAATCTCCTGTTATATTCTAGAACAGGGAATTGATTTGGGAGAGCATCAGGAAGGTGG 5585
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5521 TCAAATCTCCTGTTATATTCTAGAACAGGGAATTGATTTGGGAGAGCATCAGGAAGGTGG 5580
Qy 5586 ATGATCTGCCCAGTCACACTGTTAGTAAATTGTAGAGCCAGGACCTGAACTCTAATATAG 5645
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5581 ATGATCTGCCCAGTCACACTGTTAGTAAATTGTAGAGCCAGGACCTGAACTCTAATATAG 5640
Qy 5646 TCATGTGTTACTTAATGACGGGGACATGTTCTGAGAAATGCTTACACAAACCTAGGTGTT 5705
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5641 TCATGTGTTACTTAATGACGGGGACATGTTCTGAGAAATGCTTACACAAACCTAGGTGTT 5700
Qy 5706 GTAGCCTACTACACGCATAGGCTACATGGTATAGCCTATTGCTCCTAGACTACAAACCTG 5765
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5701 GTAGCCTACTACACGCATAGGCTACATGGTATAGCCTATTGCTCCTAGACTACAAACCTG 5760
Qy 5766 TACAGCCTGTTACTGTACTGAATACTGTGGGCAGTTGTAACACAATGGTAAGTATTTGTG 5825
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5761 TACAGCCTGTTACTGTACTGAATACTGTGGGCAGTTGTAACACAATGGTAAGTATTTGTG 5820
Qy 5826 TATCTAAACATAGAAGTTGCAGTAAAAATATGCTATTTTAATCTTATGAGACCACTGTCA 5885
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5821 TATCTAAACATAGAAGTTGCAGTAAAAATATGCTATTTTAATCTTATGAGACCACTGTCA 5880
Qy 5886 TATATACAGTCCATCATTGACCAAAACATCATATCAGCATTTTTTCTTCTAAGATTTTGG 5945
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5881 TATATACAGTCCATCATTGACCAAAACATCATATCAGCATTTTTTCTTCTAAGATTTTGG 5940
Qy 5946 GAGCACCAAAGGGATACACTAACAGGATATACTCTTTATAATGGGTTTGGAGAACTGTCT 6005
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 5941 GAGCACCAAAGGGATACACTAACAGGATATACTCTTTATAATGGGTTTGGAGAACTGTCT 6000
Qy 6006 GCAGCTACTTCTTTTAAAAAGGTGATCTACACAGTAGAAATTAGACAAGTTTGGTAATGA 6065
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6001 GCAGCTACTTCTTTTAAAAAGGTGATCTACACAGTAGAAATTAGACAAGTTTGGTAATGA 6060
Qy 6066 GATCTGCAATCCAAATAAAATAAATTCATTGCTAACCTTTTTCTTTTCTTTTCAGGTTTG 6125
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6061 GATCTGCAATCCAAATAAAATAAATTCATTGCTAACCTTTTTCTTTTCTTTTCAGGTTTG 6120
Qy 6126 AAGATGCCGCATTTGGATTGGATGAATTCCAAATTCTGCTTGCTTGCTTTTTAATATTGA 6185
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6121 AAGATGCCGCATTTGGATTGGATGAATTCCAAATTCTGCTTGCTTGCTTTTTAATATTGA 6180
Qy 6186 TATGCTTATACACTTACACTTTATGCACAAAATGTAGGGTTATAATAATGTTAACATGGA 6245
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6181 TATGCTTATACACTTACACTTTATGCACAAAATGTAGGGTTATAATAATGTTAACATGGA 6240
Qy 6246 CATGATCTTCTTTATAATTCTACTTTGAGTGCTGTCTCCATGTTTGATGTATCTGAGCAG 6305
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6241 CATGATCTTCTTTATAATTCTACTTTGAGTGCTGTCTCCATGTTTGATGTATCTGAGCAG 6300
Qy 6306 GTTGCTCCACAGGTAGCTCTAGGAGGGCTGGCAACTTAGAGGTGGGGAGCAGAGAATTCT 6365
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6301 GTTGCTCCACAGGTAGCTCTAGGAGGGCTGGCAACTTAGAGGTGGGGAGCAGAGAATTCT 6360
Qy 6366 CTTATCCAACATCAACATCTTGGTCAGATTTGAACTCTTCAATCTCTTGCACTCAAAGCT 6425
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6361 CTTATCCAACATCAACATCTTGGTCAGATTTGAACTCTTCAATCTCTTGCACTCAAAGCT 6420
Qy 6426 TGTTAAGATAGTTAAGCGTGCATAAGTTAACTTCCAATTTACATACTCTGCTTAGAATTT 6485
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6421 TGTTAAGATAGTTAAGCGTGCATAAGTTAACTTCCAATTTACATACTCTGCTTAGAATTT 6480
Qy 6486 GGGGGAAAATTTAGAAATATAATTGACAGGATTATTGGAAATTTGTTATAATGAATGAAA 6545
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6481 GGGGGAAAATTTAGAAATATAATTGACAGGATTATTGGAAATTTGTTATAATGAATGAAA 6540
Qy 6546 CATTTTGTCATATAAGATTCATATTTACTTCTTATACATTTGATAAAGTAAGGCATGGTT 6605
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6541 CATTTTGTCATATAAGATTCATATTTACTTCTTATACATTTGATAAAGTAAGGCATGGTT 6600
Qy 6606 GTGGTTAATCTGGTTTATTTTTGTTCCACAAGTTAAATAAATCATAAAACTTGA 6659
||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 6601 GTGGTTAATCTGGTTTATTTTTGTTCCACAAGTTAAATAAATCATAAAACTTGA 6654
Thus, cleavage of SEQ ID NO: 2 fulfills the requirements of instant claim 37 for a modification that eliminates the function of B2M.
It would have been prima facie obvious at the time prior to the effective filing date to eliminate the function of B2M in an engineered T cell expressing the dual CAR rendered obvious by the combined teachings of Kamb et al (‘149) and Kamb et al (‘489). One of skill in the rt would have been motivated to do so to allow for the use of allogenic T cell lines to be used to make the immune cells expressing an inhibitory receptor comprising a scFv to a HLA lost by the cancer cells by LOH , and co-expressing an activating receptor comprising an scFv to a tumor associated antigen. Further, one of skill in the art would understand that the immune cell expression of the scFv to an HLA lost by heterozygosity could result in the binding of the scFv to the other immune cells in the composition of administered immune cells and thus limit effectiveness. Thus, by eliminating the function of B2M, particularly the ethe function which controls expression of HLA and the stability of the peptide in the HLA cleft, would counter this possibility.
Double Patenting
The no statutory double patenting rejection is based on a judicially created doctrine grounded in public policy (a policy reflected in the statute) so as to prevent the unjustified or improper timewise extension of the “right to exclude” granted by a patent and to prevent possible harassment by multiple assignees. A nonstatutory double patenting rejection is appropriate where the conflicting claims are not identical, but at least one examined application claim is not patentably distinct from the reference claim(s) because the examined application claim is either anticipated by, or would have been obvious over, the reference claim(s). See, e.g., In re Berg, 140 F.3d 1428, 46 USPQ2d 1226 (Fed. Cir. 1998); In re Goodman, 11 F.3d 1046, 29 USPQ2d 2010 (Fed. Cir. 1993); In re Longi, 759 F.2d 887, 225 USPQ 645 (Fed. Cir. 1985); In re Van Ornum, 686 F.2d 937, 214 USPQ 761 (CCPA 1982); In re Vogel, 422 F.2d 438, 164 USPQ 619 (CCPA 1970); In re Thorington, 418 F.2d 528, 163 USPQ 644 (CCPA 1969).
A timely filed terminal disclaimer in compliance with 37 CFR 1.321(c) or 1.321(d) may be used to overcome an actual or provisional rejection based on nonstatutory double patenting provided the reference application or patent either is shown to be commonly owned with the examined application, or claims an invention made as a result of activities undertaken within the scope of a joint research agreement. See MPEP § 717.02 for applications subject to examination under the first inventor to file provisions of the AIA as explained in MPEP § 2159. See MPEP § 2146 et seq. for applications not subject to examination under the first inventor to file provisions of the AIA . A terminal disclaimer must be signed in compliance with 37 CFR 1.321(b).
The filing of a terminal disclaimer by itself is not a complete reply to a nonstatutory double patenting (NSDP) rejection. A complete reply requires that the terminal disclaimer be accompanied by a reply requesting reconsideration of the prior Office action. Even where the NSDP rejection is provisional the reply must be complete. See MPEP § 804, subsection I.B.1. For a reply to a non-final Office action, see 37 CFR 1.111(a). For a reply to final Office action, see 37 CFR 1.113(c). A request for reconsideration while not provided for in 37 CFR 1.113(c) may be filed after final for consideration. See MPEP §§ 706.07(e) and 714.13.
The USPTO Internet website contains terminal disclaimer forms which may be used. Please visit www.uspto.gov/patent/patents-forms. The actual filing date of the application in which the form is filed determines what form (e.g., PTO/SB/25, PTO/SB/26, PTO/AIA /25, or PTO/AIA /26) should be used. A web-based eTerminal Disclaimer may be filled out completely online using web-screens. An eTerminal Disclaimer that meets all requirements is auto-processed and approved immediately upon submission. For more information about eTerminal Disclaimers, refer to www.uspto.gov/patents/apply/applying-online/eterminal-disclaimer.
Claims 1-4 and 64 rejected on the ground of nonstatutory double patenting as being unpatentable over claims 1-27 of U.S. Patent No.11,602,544. Although the claims at issue are not identical, they are not patentably distinct from each other because the claims of the patent anticipate the instant claims.
Claim 1 of ‘544 teach a T-cell comprising (a) an activating receptor comprising an extracellular ligand binding domain specific for EGFR, and (b) an inhibitory receptor specific to HLA-A*02. The claims of the patent do not teach that the HLA-A*02 expression in a EGFR positive cancer cell due LOH
Section 804 IIb of the M.P.E.P. teaches:
The specification can be used as a dictionary to learn the meaning of a term in the patent claim. Toro Co. v. White Consol. Indus., Inc., 199 F.3d 1295, 1299, 53 USPQ2d 1065, 1067 (Fed. Cir. 1999)
In the instant case, the specification of the ‘544 patent teaches
In some embodiments, the EGFR+/HLA-A*02− cancer cell is derived from an EGFR+/HLA-A*02+ cell by loss of heterozygosity at HLA-A leading to loss of HLA-A*02.
Thus, instant claims 1-4 are obvious over the claims of the patent to the extent that the inhibitory receptor being specific for HLA-A*02 is specific for HLA-A*02 on the normal cell but lost on the tumor cell due to LOH.
Claim 9 of the patent teaches the same limitation as required in instant claim 1.
The CDR sequences of the scFv of claim 1 of the patent meet the limitations of claim 11 for CDRs selected from Table 3.
The CDR sequences of claim 4 of the patent meet the limitations of instant claim 6 for CDR sequences selected from Table 5 and the limitations of claim 7 for the CDR sequences of SEQ ID NO: 101-106. Claim 11 and 12 of the patent meet the limitations of instant claims 31 and 41. Claim 21 of the patent meets the limitations of instant claim 67. Claims 14-20 of the patent meets the limitations of instant claims 67-73. Claims 24 and 25 of the patent, drawn to a kit comprising the immune cell of claim 1 and said kit with instructions, respectively fulfills the limitations of instant claims 86 and 87 to the extent that the immune cell of claim 1 of the patent encompasses the inhibitory receptor comprising a CAR comprising a LILRB1 intracellular, hinge and transmembrane domain of claim 9 of the patent.
All other rejections and objections as set forth in the previous Office action ae withdrawn in light of applicant’s amendments.
Claims 1-14, 16, 18, 20, 21, 31-41, 59-62, 67-84 and 89-91 are rejected.
Allowable Subject Matter
Claims 42-58 are objected to as being dependent upon a rejected base claim, but would be allowable if rewritten in independent form including all of the limitations of the base claim and any intervening 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 KAREN A CANELLA whose telephone number is (571)272-0828. The examiner can normally be reached M-F 10-6:30.
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KAREN A. CANELLA
Examiner
Art Unit 1643
/Karen A. Canella/Primary Examiner, Art Unit 1643
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