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 1-21 are pending.
Applicant’s election without traverse of Group I in the reply filed on May 27, 2025 is acknowledged.
Claims 15-18 are withdrawn from further consideration by the examiner, 37 C.F.R. 1.142(b) as being drawn to non-elected inventions.
Claims 1-14 and 19-21, drawn to an isolated dimer comprising a first polypeptide and a second polypeptide, each polypeptide comprising a human IgG Fc region comprising a variant CH2 domain each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution, wherein the amino acid positions are numbered according to the EU index of Kabat, are being acted upon in this Office Action.
Priority
Applicant’ claim priority to provisional applications 61/845,948 filed July 12, 2013 and 61/671,640 filed July 13, 2012, is acknowledged.
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, or 365(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. 120 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 the first paragraph of 35 U.S.C. 112. See Transco Products, Inc. v. Performance Contracting, Inc., 38 F.3d 551, 32 USPQ2d 1077 (Fed. Cir. 1994).
The disclosure of the prior-filed application 61/671,640 fails to provide adequate support or enablement in the manner provided by the first paragraph of 35 U.S.C. 112 for one or more claims of this application. The provisional application 61/671,640 does not disclose the particular substitutions L234A, L235A, D265S as recited in claim 1, T350V, L351Y, F405A and Y407V as recited in claim 6, T350V, T366L, K392L and T394W as recited in claim 6.
Therefore for the purposes of applying prior art, the effective filing date of claims 1-14 and 19-21 is July 12, 2013, the date that provisional application 61/845,948 was filed.
Should applicant disagree with the examiner’s factual determination above, applicant should point to evidence that shows that the invention of claims 1-14 and 19-21 is in fact described in one or more of the previously-filed applications.
Information Disclosure Statement
The information disclosure statements (IDS) submitted July 9, 2024 have been considered by the examiner and an initialed copy of the IDS is included with this Office Action.
Drawings
The drawings were received on March 21, 2024. These drawings are acceptable.
Specification
The substitute specification filed on July 5, 2024 has been entered.
Applicants should amend the first line of the specification to update the relationship between the instant application, U.S. Application No. 17/207,506, file March 19, 2021, now abandoned, U.S. Application No. 16/256,824, filed January 24, 2019, now abandoned, and U.S. Application No. 13/941,449, filed July 13, 2013, now abandoned.
The lengthy specification has not been checked to the extent necessary to determine the presence of all possible minor errors. Applicant's cooperation is requested in correcting any errors of which applicant may become aware in the specification.
Claim rejections under - 35 U.S.C. 112
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 1-14 and 19-21 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 pre-AIA the inventor(s), at the time the application was filed, had possession of the claimed invention.
MPEP § 2163 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.
Claim 1 encompasses any isolated dimer comprising a first polypeptide and a second polypeptide, each polypeptide comprising a human IgG Fc region comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution, wherein the amino acid positions are numbered according to the EU index of Kabat.
Claim 2 encompasses the isolated dimer of claim 1, wherein at least one of the first and second polypeptides further comprises an antigen-binding domain.
Claim 3 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises an scFv.
Claim 4 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises a Fab.
Claim 5 encompasses the isolated dimer of claim 1, wherein one of the first and second polypeptides further comprises an scFv and the other of the polypeptides further comprises a Fab.
Claim 6 encompasses the isolated dimer of claim 1, wherein one of the first and second polypeptides further comprises amino acid substitutions T350V, L351Y, F405A, and Y407V, and the other of the polypeptides further comprises amino acid substitutions T350V, T366L, K392L, and T394W.
Claim 7 encompasses the isolated dimer of claim 1, wherein the IgG is an IgG1.
Claim 8 encompasses any antibody comprising the isolated dimer of claim 1.
Claim 9 encompasses the e antibody of claim 8, wherein the antibody is any monoclonal antibody, any humanized antibody, or any human antibody.
Claim 10 encompasses the antibody of claim 8, wherein the antibody is multispecific.
Claim 11 encompasses the antibody of claim 8, wherein the antibody is bispecific.
Claim 12 encompasses the isolated dimer of claim 1, wherein the dimer is a heterodimer.
Claim 13 encompasses a pharmaceutical composition comprising the isolated dimer of claim 1 and a pharmaceutically acceptable carrier.
Claim 14 encompasses a pharmaceutical composition comprising the antibody of claim 8 and a pharmaceutically acceptable carrier.
Claim 19 encompasses the isolated dimer of claim 12, wherein each of the first and second polypeptides further comprises an antigen-binding domain.
Claim 20 encompasses the isolated dimer of claim 19, wherein each antigen-binding domain binds a different antigen.
Claim 21 encompasses the isolated dimer of claim 20, wherein one of the antigen-binding domains binds to a CD3 complex.
The specification discloses heterodimer:
[0359] v4541 has an anti-CD3 BiTE™ (VH-VL) 44-100SS scFv on chain A and anti-CD19 BiTE™ (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 50 and 52] Cyno/Human Cross-Reactive Anti-CD3 and Anti-CD19 Bispecific Fc Knock-Out Variants with or without Disulfide 44-100 Stabilization
[0360] v4542 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 54 and 56].
[0361] v4543 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 58 and 60]
[0362] v4544 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 62 and 64]
[0363] v4545 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 66 and 68]
[0364] v4546 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 70 and 72]
[0365] v4547 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 74 and 76].
[0366] v4548 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 78 and 80] v4549 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 82 and 84]
Regarding multispecific antibody, the specification discloses:
Example 3: Heteromultimer v873 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 4: Heteromultimers Bind Selectively to CD3- and CD19-Expressing Cells
[0384] The ability of an exemplary heteromultimer, v873, to bind specifically to CD3 and CD19 was assessed by FACS. One-armed antibodies (OAAs) against CD3 and CD19 were also prepared as described in Example 2 and tested as controls in the whole cell FACS binding assay described below.
Example 5: Heteromultimers Mediate PBMC Killing of Target Raji B Cells
Example 6: Heteromultimers Mediate Redirected Killing of Target Raji B Cells with Resting and IL-2 Activated CD4+ and CD8+ T Cells
[0405] The ability of an exemplary heteromultimers, v875, v1379, v1380 to mediate CD4+ and CD8+ T cell cytoxicity against target Raji B-cells was measured as described in Example 5.
Example 7: The Heterodimeric Fc Contributes to Target Raji B Cell Cytotoxicity
[0410] The ability of exemplary heteromultimers, v875, and v873, to mediate target Raji B-cell cytotoxicity in the presence and absence of Fc was measured as described in Example 5.
Example 8: Heteromultimers Mediate Autologous B Cell Cytotoxicity
[0413] The ability of exemplary the heteromultimer v875 and v873 to kill autologous B-cells was measured in total and resting IL-2-stimulated PBMCs where the percent of CD19+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
Example 9: Heteromultimer v875 Spares Autologous T Cell Cytotoxicity Compared to BiTE
[0415] The effects of exemplary heteromultimer v875 and v873 treatment on the autologous T cell population was assessed in total and resting IL-2-stimulated PBMCs where the percent of CD3+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
[0416] FIG. 22 shows, relative to untreated media and human IgG controls, v875 has a more selective B cell killing by sparing more autologous T cells compared to v873 and v891.
Example 10: Design, Expression and Purification of Heteromultimers with an Albumin Scaffold
[0417] The following exemplary CD3-CD19 binding heteromultimers based on an albumin scaffold were designed and prepared as follows.
[0418] The sequences for the anti-CD19 and anti-CD3 scFvs were chosen from two molecules that are currently in clinical trials and are well documented and tested for stability and production. The anti-CD19 and anti-CD3 scFv were directly adopted from the BiTE molecule blinatumomab. The antiCD3 scFv was chosen in the VH-VL orientation, consistent with what used in BiTE. The benchmark molecule was an scFv molecule based on BiTE (v891). AlbuCORE_1 (ABH2) CD3/CD19 fusions were created by attaching the antiCD3 warhead to the natural N terminus of fragment 1 and the antiCD19 to the C terminus of fragment 2 (v1092, polypeptide sequences corresponding to SEQ ID NO:264 and 266). The linkers used were identical to the ones used for the multivalent HER2 AlbuCORE experiments: GGGS (SEQ ID NO: 278) at the N terminus of fragment 1 and (GGSG) 4GG (SEQ ID NO: 279) at the C terminus of fragment 2. A second molecule was created where the warheads were reversed (i.e. anti-CD19 warhead at the natural N terminus of fragment 1 and the anti-CD3 at the C terminus of fragment 2, v1093. v1094 was designed to accommodate two different fusions at the natural termini of the albumin polypeptide (polypeptide sequences corresponding to SEQ ID NO:268). The scFv fusions were linked to the albumin molecule through a GGS linker at the N terminus and a GGSG (SEQ ID NO: 280) linker at the C terminus. The length of the linkers reflect the ones used in the MM-111 molecule, despite having a different sequence type.
Example 11: Heteromultimers with an Albumin Scaffold Bind Specifically to CD3- or CD19-Expressing Cells
[0421] The ability of Anti-CD3×CD19 loaded AlbuCORE-1 (v1092) to CD3+ and CD19+ cells was assessed using FACS and compared to WT-HSA loaded with the same anti-CD scFvs (v1094).
[0422] The results are shown in FIG. 4 and demonstrate that both v1092 and v1094 are able to bind to CD3-expressing Jurkat T-cells and to CD19-expressing Raji B-cells.
Example 12: Heteromultimers with Heterodimeric Fc or Albumin Scaffolds Show Comparable B-Cell Targeting and T-Cell Bridging
Example 13: Exemplary Heteromultimers have Higher Anti-CD3 K.SUB.D .and Higher Bmax in Binding to T and B Cell as Determined by FACS
Example 14: Heteromultimer v875 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 15: Bridging of B and T Cells by Heteromultimers is Robust at Varying Antibody Concentrations or Cell Ratios
Example 16: Bridging of B and T Cells is Robust Across Differently Engineered Heteromultimers Constructs
Example 17: Effects of v875, v1380, v1379 on IL-2 Activated and Resting CD20+, CD4+, CD8+Subsets
Example 18: Exemplary Heteromultimers v875 and v873 Require the Presence of Both Effector T Cell and Target B Cells to Mediate Cytotoxic Effects
Example 19: Exemplary Heteromultimers can Mediate ADCC or Impaired ADCC to Target Daudi B Cells
[0444] Antibody-dependent cell-mediated cytotoxicity assays (ADCC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells and FcRy3a immobilized NK92 cells as the effector cells (GS193761) by the following method.
Example 20: Exemplary Heteromultimers have Impaired CDC-Mediated Lysis of Daudi B Cells
[0449] Cell based complement dependent cytotoxicity assays (CDC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells. Human serum from healthy donors (NHS) was used as the source of complement. 10 μl NHS (10% final concentration in 40 reaction volume) were added to each well to initiate the CDC cascade and incubated for 2 hours. Cell viability was measured with CellTiter-Glo@ Luminescent Cell Viability Assay Kit.
Example 21: Cell Proliferation and Cytokine Release Assessment of Exemplary Heteromultimers
Example 22: Exemplary Heteromultimers can Bridge Two or More Target B Cells Per Effector T Cell
[0465] The ability and ratio of numbers of T cell bridged to B cells was examined with the exemplary heteromultimer v875 by microscopy using the method described in Example 3 with the following modifications.
[0466] Labeled Raji B cells (red) and labeled Jurkat T cells (blue) were incubated for 30 min at RT with 3 nM of human IgG or v875. The cell suspension was concentrated by removing 180 μl of supernatant. Cell were resuspended in the remaining volume and imaged at 200× and 400×.
[0467] FIG. 30 shows the results from the T:B cell bridging microscopy comparing v875 and human IgG (3 nM) at 200× and 400× magnification; the phase image (top panel), fluorescence image (middle panel) and inverted fluorescence (bottom panel) are presented. FIG. 30A shows a direct comparison of human IgG and v875 at 200× magnification and illustrates a higher amount of bridging visible between Raji B cell and Jurkat T cells compared to human IgG. FIG. 30B and FIG. 30C show two fields of view for v875 (FIG. 30B) and human IgG (FIG. 30C) at 400× magnification. FIG. 30B shows images of v875-mediated immune complex formation between Jurkat T cells (dark grey cells in fluorescence inverted image) and Raji B cells (light grey cells in fluorescence inverted image), and that one Jurkat T cell can bridge 1-3 Raji B cells. FIG. 30C shows images following human IgG incubation with Jurkat T cells and Raji B cells. FIG. 30C shows an absence of the Jurkat-Raji bridging following incubation with human IgG, compared to the v875-mediated Jurkat: Raji bridging that is visible in FIG. 30B.
Example 23: Exemplary Heteromultimer Binding to Fcγ Receptors as Assessed by Surface Plasmon Resonance
Example 25: Expression and Purification of Heteromultimers
[0476] Description of the methods used in the expression and purification of exemplary heteromultimer are described in Example 2.
Example 26: Exemplary Heteromultimers can be Purified to >99% Heterodimer Purity and <1% Aggregates
Example 27: Exemplary Heteromultimers have a CH3 Tm that is Greater than 75° C.
[0487] CH3 domain stability of exemplary heteromultimers was examined by DSC using the following method. All DSC experiments were carried out using a GE VP-Capillary instrument. The proteins were buffer-exchanged into PBS (pH 7.4) and diluted to 0.3 to 0.7 mg/mL with 0.137 mL loaded into the sample cell and measured with a scan rate of 1° C./min from 20 to 100° C. Data was analyzed using the Origin software (GE Healthcare) with the PBS buffer background subtracted.
[0488] The DSC results shown in FIGS. 33A, B and C show that v875 has an estimated CH3 Tm >76° C. (FIG. 33A), v1380 has an estimated CH3 Tm >82.3° C. (FIG. 33B), and v1379 has an estimated CH3 Tm >82.5° C. (FIG. 33C).
Example 28: Design, Expression and Purification of CD3/CD20 and Additional CD3/CD19 Heteromultimer Constructs
[0489] V5850 (corresponding to polypeptide sequences SEQ ID NOs: 203, 205 and 207), v5851 (corresponding to polypeptide sequences SEQ ID NOs: 209, 211 and 213), v5852 (corresponding to polypeptide sequences SEQ ID NOs: 215, 217 and 219), v6324 (corresponding to polypeptide sequences SEQ ID NOs: 221 and 223), v6325 (corresponding to polypeptide sequences SEQ ID NOs: 225, 227 and 229), v1813 (corresponding to polypeptide sequences SEQ ID NOs: 231, 233 and 235), v1821 (corresponding to polypeptide sequences SEQ ID NOs: 237, 239 and 241), v1823 (corresponding to polypeptide sequences SEQ ID NOs: 243, 245 and 247) exemplify bispecific CD3/CD19 or CD3/CD20 hybrid heterodimeric Fc constructs. Bispecific hybrid variants are composed of a F(ab′) on either chain A or B paired with an scFv-Fc on the alternate polypeptide chain. Chain A of the heterodimer Fc is comprised of the following mutations: T350V_L351Y_F405A_Y407V and Chain B of the heterodimer Fc is comprised of the following mutations: T350V_T366L_K392L_T394W. v6324 exemplifies bispecific CD3/CD20 scFv heterodimeric Fc constructs. V1813, v1821, and v1823 exemplify CD3/CD20 common light chain heterodimeric Fc constructs. Common light chain variants are composed of two different F(ab′) s, each on complimentary heterodimer Fc, which share a single light chain. The specific variant composition is indicated in Table 7.
However, the disclosure of just heterodimer is not representative of all dimer, e.g., homodimer or heterodimer comprises a first human IgG Fc and a second human IgG Fc wherein each Fc comprises a D265S, a L234A and L235A.
Regarding antibody, the specification does not describe the structure, e.g., amino acid sequence of the heavy and light chain variable region of all antigen-binding domain, e.g., any scFv (claim 3), Fab (claim 4), antibody (claim 8), (humanized or human or monoclonal antibody (claim 9), multispecific antibody (claim 10) or bispecific antibody (claim 11) that correlated with binding to any and all different antigen (claim 20) and all CD3 complex (claim 21) and pharmaceutical composition (claims 13-14) to enable one of skilled in the art to visualize or recognize the member of the genus of the actual claimed dimer and antibody themselves. The specification does not describe a representative number of species of dimer or antibody as a pharmaceutical composition for treating which disease. The specification does not describe the structure correlated with binding falling within the scope of the genus or structural common to the members of the genus so the one of skill in the art can visualize or recognize the member of the genus. An adequate written description must contain enough information about the actual makeup of the claimed products – “a precise definition, such as structure, formula, chemic name, physical properties of other properties, of species falling with the genus sufficient to distinguish the gene from other materials”, which may be present in “functional terminology when the art has established a correlation between structure and function” (Amgen page 1361).
Notably, the specification, does not describe the structure, e.g., amino acid sequence of the heavy and light chain variable domains or the six CDRs that correlated with binding to any antigen (claim 20) and any CD3 complex (claim 21). There is no limitation on the structure and the antigen to which it binds.
At the time of the invention was made, it is known in the art that antibodies have a large repertoire of distinct structures and that a huge variety of antibodies can be made to bind to a single epitope.
For example, Lloyd et al. taught that hundreds of functional antibody fragments can be isolated from an antibody library that bind to the same antigen wherein these antibodies have distinct heavy and light chain sequences (Lloyd et al. Protein Engineering, Design & Selection 22:159-168, 2009, PTO 1449; see, e.g., Discussion).
Similarly, Edwards et al., (J Mol Biol. 334(1): 103-118, 2003; PTO 1449), found that over 1000 antibodies, all different in amino acid sequence, were generated to a single protein; 568 different amino acid sequences identified for the V(H) CDR3 domains of these antibodies (Abstract).
Poosarla et al (Biotechn. Bioeng., 114(6): 1331-1342, 2017; PTO 892) teach substantial diversity in designed mAbs (sharing less than 75% sequence similarity to all existing natural antibody sequences) that bind to the same 12-mer peptide, binding to different epitopes on the same peptide. Said reference further teaches “most B-cell epitopes... in nature consist of residues from different regions of the sequence and are discontinuous...de novo antibody designs against discontinuous epitopes present additional challenge.
Even assuming the antibody binds to CD3, Shepard et al (PLOSOne 18(6): e0273884, June 22, 2023; PTO 892) teach the unpredictability of swapping CD3 binding domains in the context of a BiTE in so far as the binding requirement to CD3 epsilon much less the extracellular epitope:
“We next sought to test our platform for flexibility with respect to screening novel T-cell engaging domains of the bispecific molecule. We developed a number of novel mouse monoclonal antibodies (mAbs) against human CD3 complex using a multi-antigen immunization strategy in mice and traditional hybridoma screening. Through this process, we were able to identify several monoclonal antibodies with reactivity to Jurkat cells (Fig 4A) and human T cells (Fig 4B). To assess whether scFvs derived from novel CD3-targeted mAbs would be functional within as part of a BiTE molecule, we cloned 4 anti-human CD3 single chain variable fragments into CD19 or EGFRvIII specific BiTE plasmids (Fig 4C). We then generated supernatants using transient transfection of HEK293T as described above. BiTE supernatants were screened for activity using Jurkat cells in co-culture with EGFRvIII-expressing U87-VIII targets or CD19-expressing Raji cells. Previously tested constructs using OKT3 CD3-engager arms showed activity with both EGFRvIII and CD19 specific BiTEs, whereas we detected activity for only one of our novel CD3-engager BiTEs and only when combined with an EGFRvIII-specific scFv (Fig 4D). To confirm these results, we repeated BiTE production and Jurkat co-culture screening of CD19 and EGFRvIII targeted BiTE molecules incorporating OKT3 or the novel 1E2 CD3-targeting single chain variable fragment. Whereas EGFRvIII BiTEs incorporating an scFv derived from the 1E2 mAb or OKT3 showed specific reactivity against EGFRvIII expressing U87-vIII cells, only CD19-OKT3 showed reactivity to CD19-expressing Raji cells (Fig 4E). These results indicate that the novel CD3-targeting 1E2-scFv is functional only for an EGFRvIII-targeting BiTE but not a CD19-targeting BiTE, likely due to the specific binding characteristics of the CD19 or EGFRvIII scFv elements.”
“Similarly, we also find that only one of the novel CD3-targeting scFvs tested here showed activity in BiTE format, and even then, activity was restricted to combination with EGFRvIII-targeting scFv and not with a CD19-specific scFv. Again, we have no insight into the failure rate for BiTE molecules in this assay, as the intent of this assay is to provide a rapid means for testing biological activity rather than focusing on various aspects of antibody characterization. We are currently undertaking molecular studies to test whether incorporating different linker domains may be able to improve the activity of BiTEs using the novel CD3-scFv reported to have activity here, something that may provide further insight into the design constraints for these BiTE molecules.”
Given that hundreds of unique antibody structures may bind a single antigen, the structure of an antibody cannot be predicted from the structure of the antigen (as held in Amgen), and a single species, or small group of species, cannot define a structure-function relationship so as to be representative of all the antibodies that bind to that antigen (as held in Abbvie).
Although one of skill in the art could make a mAb against an antigen/CD3 complex by screening a human antibody phage display library, test candidates, and produce a corresponding antibody, note that: “Possession may not be shown by merely describing how to obtain possession of members of the claimed genus or how to identify their common structural features.” See University of Rochester, 358 F.3d at 927, 69 USPQ2d at 1895.
Vas-Cath Inc. v. Mahurkar, 19 USPQ2d 1111, makes clear that “applicant must convey with reasonable clarity to those skilled in the art that, as of the filing date sought, he or she was in possession of the invention. The invention is, for purposes of the written description inquiry, whatever is now claimed.” (See page 1117.) The specification does not “clearly allow persons of ordinary skill in the art to recognize that [he or she] invented what is claimed.” (See Vas-Cath at page 1116.).
Adequate written description requires more than a mere statement that it is part of the invention and reference to a potential method for isolating it. See Fiers v. Revel, 25 USPQ2d 1601, 1606 (CAFC 1993) and Amgen Inc. v. Chugai Pharmaceutical Co. Ltd., 18 USPQ2d 1016.
One cannot describe what one has not conceived. See Fiddles v. Baird, 30 USPQ2d 1481, 1483. In Fiddles v. Baird, claims directed to mammalian FGF’s were found unpatentable due to lack of written description for the broad class. The specification provided only the bovine sequence. Thus, the specification fails to describe these DNA sequences.
For genus claims, an adequate written description of a claimed genus requires more than a generic statement of an invention's boundaries. A patent must set forth either a representative number of species falling within the scope of the genus or structural features common to the members of the genus. Kubin, Exparte, 83 USPQ2d 1410 (Bd. Pat. App. & Int. 2007); Ariad Pharms., Inc. v. Eli Lilly& Co., 598 F.3d 1336, 1350 (Fed. Cir. 2010).
Therefore, only a bispecific CD19 and CD3 scFvs fused to an asymmetric IgG1 heterodimer comprising a first Fc polypeptide and a second Fc polypeptide, each Fc polypeptide comprising a human CH2 domain, each CH2 domain comprises a L234A amino acid substitution, a L235A amino acid substitution and a D265S amino acid substitution, wherein the amino acid positions are numbered according to the EU index of Kabat, wherein the first and second polypeptides comprise the amino acid sequences selected from the group consisting of SEQ ID NO: 38 and SEQ ID NO: 40, SEQ ID NO: 42 and SEQ ID NO: 44, SEQ ID NO: 46 and SEQ ID NO: 48, SEQ ID NO: 66 and SEQ NO: 68, SEQ ID NO: 70 and SEQ NO: 72, SEQ ID NO: 74 and SEQ NO: 76, SEQ ID NO: 78 and SEQ NO: 80, SEQ ID NO: 82 and SEQ NO: 84, SEQ ID NO: 133 and SEQ ID NO: 134, SEQ ID NO: 137 and SEQ ID NO: 138, and SEQ ID NO: 141 and SEQ ID NO: 142, but not the full breadth of the claims meets the written description provision of 35 U.S.C. § 112, first paragraph.
Applicant is reminded that Vas-Cath makes clear that the written description provision of 35 U.S.C. § 112 is severable from its enablement provision (see page 1115).
Claims 1-14 and 19-21 are rejected under 35 U.S.C. 112(a) or 35 U.S.C. 112 (pre-AIA ), first paragraph, because the specification, while being enabling for a bispecific CD19 and CD3 scFvs fused to an asymmetric IgG1 heterodimer comprising a first Fc polypeptide and a second Fc polypeptide, each Fc polypeptide comprising a human CH2 domain, each CH2 domain comprises a L234A amino acid substitution, a L235A amino acid substitution and a D265S amino acid substitution, wherein the amino acid positions are numbered according to the EU index of Kabat, wherein the first and second polypeptides comprise the amino acid sequences selected from the group consisting of SEQ ID NO: 38 and SEQ ID NO: 40, SEQ ID NO: 42 and SEQ ID NO: 44, SEQ ID NO: 46 and SEQ ID NO: 48, SEQ ID NO: 66 and SEQ NO: 68, SEQ ID NO: 70 and SEQ NO: 72, SEQ ID NO: 74 and SEQ NO: 76, SEQ ID NO: 78 and SEQ NO: 80, SEQ ID NO: 82 and SEQ NO: 84, SEQ ID NO: 133 and SEQ ID NO: 134, SEQ ID NO: 137 and SEQ ID NO: 138, and SEQ ID NO: 141 and SEQ ID NO: 142, does not reasonably provide enablement for the isolated dimer set forth in claims 1-14 and 19-21. The specification does not enable any person skilled in the art to which it pertains, or with which it is most nearly connected, to make/use the invention commensurate in scope with these claims.
Claim 1 encompasses any isolated dimer comprising a first polypeptide and a second polypeptide, each polypeptide comprising a human IgG Fc region comprising a variant CH2 domain, each variant CH2 domain comprising a L234A amino acid substitution, a L235A amino acid substitution, and a D265S amino acid substitution, wherein the amino acid positions are numbered according to the EU index of Kabat.
Claim 2 encompasses the isolated dimer of claim 1, wherein at least one of the first and second polypeptides further comprises an antigen-binding domain.
Claim 3 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises an scFv.
Claim 4 encompasses the isolated dimer of claim 2, wherein the antigen-binding domain comprises a Fab.
Claim 5 encompasses the isolated dimer of claim 1, wherein one of the first and second polypeptides further comprises an scFv and the other of the polypeptides further comprises a Fab.
Claim 6 encompasses the isolated dimer of claim 1, wherein one of the first and second polypeptides further comprises amino acid substitutions T350V, L351Y, F405A, and Y407V, and the other of the polypeptides further comprises amino acid substitutions T350V, T366L, K392L, and T394W.
Claim 7 encompasses the isolated dimer of claim 1, wherein the IgG is an IgG1.
Claim 8 encompasses any antibody comprising the isolated dimer of claim 1.
Claim 9 encompasses the e antibody of claim 8, wherein the antibody is any monoclonal antibody, any humanized antibody, or any human antibody.
Claim 10 encompasses the antibody of claim 8, wherein the antibody is multispecific.
Claim 11 encompasses the antibody of claim 8, wherein the antibody is bispecific.
Claim 12 encompasses the isolated dimer of claim 1, wherein the dimer is a heterodimer.
Claim 13 encompasses a pharmaceutical composition comprising the isolated dimer of claim 1 and a pharmaceutically acceptable carrier.
Claim 14 encompasses a pharmaceutical composition comprising the antibody of claim 8 and a pharmaceutically acceptable carrier.
Claim 19 encompasses the isolated dimer of claim 12, wherein each of the first and second polypeptides further comprises an antigen-binding domain.
Claim 20 encompasses the isolated dimer of claim 19, wherein each antigen-binding domain binds a different antigen.
Claim 21 encompasses the isolated dimer of claim 20, wherein one of the antigen-binding domains binds to a CD3 complex.
The specification discloses heterodimer:
[0359] v4541 has an anti-CD3 BiTE™ (VH-VL) 44-100SS scFv on chain A and anti-CD19 BiTE™ (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 50 and 52] Cyno/Human Cross-Reactive Anti-CD3 and Anti-CD19 Bispecific Fc Knock-Out Variants with or without Disulfide 44-100 Stabilization
[0360] v4542 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 54 and 56].
[0361] v4543 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 58 and 60]
[0362] v4544 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 62 and 64]
[0363] v4545 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MOR208 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 66 and 68]
[0364] v4546 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 70 and 72]
[0365] v4547 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VH-VL) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 74 and 76].
[0366] v4548 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 78 and 80] v4549 has a cyno/human cross-reactive anti-CD3 BiTE™ 12C (VH-VL) 44-100SS scFv on chain A and cyno/human cross-reactive anti-CD19 MDX-1342 (VL-VH) 44-100SS scFv on chain B of the heterodimer Fc with the following mutations D265S_L234A_L235A_T350V_L351Y_F405A_Y407V on chain A and D265S_L234A_L235A_T350V_T366L_K392L_T394W on chain B. [Polypeptide sequences correspond to SEQ ID No: 82 and 84]
Regarding multispecific antibody, the specification discloses:
Example 3: Heteromultimer v873 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 4: Heteromultimers Bind Selectively to CD3- and CD19-Expressing Cells
[0384] The ability of an exemplary heteromultimer, v873, to bind specifically to CD3 and CD19 was assessed by FACS. One-armed antibodies (OAAs) against CD3 and CD19 were also prepared as described in Example 2 and tested as controls in the whole cell FACS binding assay described below.
Example 5: Heteromultimers Mediate PBMC Killing of Target Raji B Cells
Example 6: Heteromultimers Mediate Redirected Killing of Target Raji B Cells with Resting and IL-2 Activated CD4+ and CD8+ T Cells
[0405] The ability of an exemplary heteromultimers, v875, v1379, v1380 to mediate CD4+ and CD8+ T cell cytoxicity against target Raji B-cells was measured as described in Example 5.
Example 7: The Heterodimeric Fc Contributes to Target Raji B Cell Cytotoxicity
[0410] The ability of exemplary heteromultimers, v875, and v873, to mediate target Raji B-cell cytotoxicity in the presence and absence of Fc was measured as described in Example 5.
Example 8: Heteromultimers Mediate Autologous B Cell Cytotoxicity
[0413] The ability of exemplary the heteromultimer v875 and v873 to kill autologous B-cells was measured in total and resting IL-2-stimulated PBMCs where the percent of CD19+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
Example 9: Heteromultimer v875 Spares Autologous T Cell Cytotoxicity Compared to BiTE
[0415] The effects of exemplary heteromultimer v875 and v873 treatment on the autologous T cell population was assessed in total and resting IL-2-stimulated PBMCs where the percent of CD3+ 7AAD+ cells following incubation with v875 and v873 (300 nM, n=3 donors) was determined by flow cytometry as described in Example 5.
[0416] FIG. 22 shows, relative to untreated media and human IgG controls, v875 has a more selective B cell killing by sparing more autologous T cells compared to v873 and v891.
Example 10: Design, Expression and Purification of Heteromultimers with an Albumin Scaffold
[0417] The following exemplary CD3-CD19 binding heteromultimers based on an albumin scaffold were designed and prepared as follows.
[0418] The sequences for the anti-CD19 and anti-CD3 scFvs were chosen from two molecules that are currently in clinical trials and are well documented and tested for stability and production. The anti-CD19 and anti-CD3 scFv were directly adopted from the BiTE molecule blinatumomab. The antiCD3 scFv was chosen in the VH-VL orientation, consistent with what used in BiTE. The benchmark molecule was an scFv molecule based on BiTE (v891). AlbuCORE_1 (ABH2) CD3/CD19 fusions were created by attaching the antiCD3 warhead to the natural N terminus of fragment 1 and the antiCD19 to the C terminus of fragment 2 (v1092, polypeptide sequences corresponding to SEQ ID NO:264 and 266). The linkers used were identical to the ones used for the multivalent HER2 AlbuCORE experiments: GGGS (SEQ ID NO: 278) at the N terminus of fragment 1 and (GGSG) 4GG (SEQ ID NO: 279) at the C terminus of fragment 2. A second molecule was created where the warheads were reversed (i.e. anti-CD19 warhead at the natural N terminus of fragment 1 and the anti-CD3 at the C terminus of fragment 2, v1093. v1094 was designed to accommodate two different fusions at the natural termini of the albumin polypeptide (polypeptide sequences corresponding to SEQ ID NO:268). The scFv fusions were linked to the albumin molecule through a GGS linker at the N terminus and a GGSG (SEQ ID NO: 280) linker at the C terminus. The length of the linkers reflect the ones used in the MM-111 molecule, despite having a different sequence type.
Example 11: Heteromultimers with an Albumin Scaffold Bind Specifically to CD3- or CD19-Expressing Cells
[0421] The ability of Anti-CD3×CD19 loaded AlbuCORE-1 (v1092) to CD3+ and CD19+ cells was assessed using FACS and compared to WT-HSA loaded with the same anti-CD scFvs (v1094).
[0422] The results are shown in FIG. 4 and demonstrate that both v1092 and v1094 are able to bind to CD3-expressing Jurkat T-cells and to CD19-expressing Raji B-cells.
Example 12: Heteromultimers with Heterodimeric Fc or Albumin Scaffolds Show Comparable B-Cell Targeting and T-Cell Bridging
Example 13: Exemplary Heteromultimers have Higher Anti-CD3 K.SUB.D .and Higher Bmax in Binding to T and B Cell as Determined by FACS
Example 14: Heteromultimer v875 is Able to Bridge Jurkat CD3 T Cells and Raji CD19 B Cells
Example 15: Bridging of B and T Cells by Heteromultimers is Robust at Varying Antibody Concentrations or Cell Ratios
Example 16: Bridging of B and T Cells is Robust Across Differently Engineered Heteromultimers Constructs
Example 17: Effects of v875, v1380, v1379 on IL-2 Activated and Resting CD20+, CD4+, CD8+Subsets
Example 18: Exemplary Heteromultimers v875 and v873 Require the Presence of Both Effector T Cell and Target B Cells to Mediate Cytotoxic Effects
Example 19: Exemplary Heteromultimers can Mediate ADCC or Impaired ADCC to Target Daudi B Cells
[0444] Antibody-dependent cell-mediated cytotoxicity assays (ADCC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells and FcRy3a immobilized NK92 cells as the effector cells (GS193761) by the following method.
Example 20: Exemplary Heteromultimers have Impaired CDC-Mediated Lysis of Daudi B Cells
[0449] Cell based complement dependent cytotoxicity assays (CDC) were performed with v875, v1379 and v1380 using Daudi cells as target B cells. Human serum from healthy donors (NHS) was used as the source of complement. 10 μl NHS (10% final concentration in 40 reaction volume) were added to each well to initiate the CDC cascade and incubated for 2 hours. Cell viability was measured with CellTiter-Glo@ Luminescent Cell Viability Assay Kit.
Example 21: Cell Proliferation and Cytokine Release Assessment of Exemplary Heteromultimers
Example 22: Exemplary Heteromultimers can Bridge Two or More Target B Cells Per Effector T Cell
[0465] The ability and ratio of numbers of T cell bridged to B cells was examined with the exemplary heteromultimer v875 by microscopy using the method described in Example 3 with the following modifications.
[0466] Labeled Raji B cells (red) and labeled Jurkat T cells (blue) were incubated for 30 min at RT with 3 nM of human IgG or v875. The cell suspension was concentrated by removing 180 μl of supernatant. Cell were resuspended in the remaining volume and imaged at 200× and 400×.
[0467] FIG. 30 shows the results from the T:B cell bridging microscopy comparing v875 and human IgG (3 nM) at 200× and 400× magnification; the phase image (top panel), fluorescence image (middle panel) and inverted fluorescence (bottom panel) are presented. FIG. 30A shows a direct comparison of human IgG and v875 at 200× magnification and illustrates a higher amount of bridging visible between Raji B cell and Jurkat T cells compared to human IgG. FIG. 30B and FIG. 30C show two fields of view for v875 (FIG. 30B) and human IgG (FIG. 30C) at 400× magnification. FIG. 30B shows images of v875-mediated immune complex formation between Jurkat T cells (dark grey cells in fluorescence inverted image) and Raji B cells (light grey cells in fluorescence inverted image), and that one Jurkat T cell can bridge 1-3 Raji B cells. FIG. 30C shows images following human IgG incubation with Jurkat T cells and Raji B cells. FIG. 30C shows an absence of the Jurkat-Raji bridging following incubation with human IgG, compared to the v875-mediated Jurkat: Raji bridging that is visible in FIG. 30B.
Example 23: Exemplary Heteromultimer Binding to Fcγ Receptors as Assessed by Surface Plasmon Resonance
Example 25: Expression and Purification of Heteromultimers
[0476] Description of the methods used in the expression and purification of exemplary heteromultimer are described in Example 2.
Example 26: Exemplary Heteromultimers can be Purified to >99% Heterodimer Purity and <1% Aggregates
Example 27: Exemplary Heteromultimers have a CH3 Tm that is Greater than 75° C.
[0487] CH3 domain stability of exemplary heteromultimers was examined by DSC using the following method. All DSC experiments were carried out using a GE VP-Capillary instrument. The proteins were buffer-exchanged into PBS (pH 7.4) and diluted to 0.3 to 0.7 mg/mL with 0.137 mL loaded into the sample cell and measured with a scan rate of 1° C./min from 20 to 100° C. Data was analyzed using the Origin software (GE Healthcare) with the PBS buffer background subtracted.
[0488] The DSC results shown in FIGS. 33A, B and C show that v875 has an estimated CH3 Tm >76° C. (FIG. 33A), v1380 has an estimated CH3 Tm >82.3° C. (FIG. 33B), and v1379 has an estimated CH3 Tm >82.5° C. (FIG. 33C).
Example 28: Design, Expression and Purification of CD3/CD20 and Additional CD3/CD19 Heteromultimer Constructs
[0489] V5850 (corresponding to polypeptide sequences SEQ ID NOs: 203, 205 and 207), v5851 (corresponding to polypeptide sequences SEQ ID NOs: 209, 211 and 213), v5852 (corresponding to polypeptide sequences SEQ ID NOs: 215, 217 and 219), v6324 (corresponding to polypeptide sequences SEQ ID NOs: 221 and 223), v6325 (corresponding to polypeptide sequences SEQ ID NOs: 225, 227 and 229), v1813 (corresponding to polypeptide sequences SEQ ID NOs: 231, 233 and 235), v1821 (corresponding to polypeptide sequences SEQ ID NOs: 237, 239 and 241), v1823 (corresponding to polypeptide sequences SEQ ID NOs: 243, 245 and 247) exemplify bispecific CD3/CD19 or CD3/CD20 hybrid heterodimeric Fc constructs. Bispecific hybrid variants are composed of a F(ab′) on either chain A or B paired with an scFv-Fc on the alternate polypeptide chain. Chain A of the heterodimer Fc is comprised of the following mutations: T350V_L351Y_F405A_Y407V and Chain B of the heterodimer Fc is comprised of the following mutations: T350V_T366L_K392L_T394W. v6324 exemplifies bispecific CD3/CD20 scFv heterodimeric Fc constructs. V1813, v1821, and v1823 exemplify CD3/CD20 common light chain heterodimeric Fc constructs. Common light chain variants are composed of two different F(ab′) s, each on complimentary heterodimer Fc, which share a single light chain. The specific variant composition is indicated in Table 7.
However, the specification does not teach dimer, e.g., homodimer comprises a first human IgG Fc and a second human IgG Fc wherein each Fc comprises a D265S, a L234A and L235A.
Regarding antibody