T cell redirecting bispecific antibodies against cytomegaloviruses

Abstract

Disclosed herein T-cell redirecting bispecific antibodies (TRBAs) against cytomegalovirus (CMV) and compositions and methods of making and using thereof.

Description

T CELL REDIRECTING BISPECIFIC ANTIBODIES AGAINST CYTOMEGALOVIRUSES

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Patent Application No. 63 / 504,760, filed May 29, 2023, which is herein incorporated by reference in its entirety.

[0003] REFERENCE TO A SEQUENCE LISTING SUBMITTED VIA PATENT CENTER

[0004] The content of the XML file of the sequence listing named“20240510_034044_247W01_ST26” which is 45,163 bytes in size was created on May 10, 2024 and electronically submitted via Patent Center herewith the application is incorporated herein by reference in its entirety.

[0005] BACKGROUND OF THE INVENTION

[0006] 1. FIELD OF THE INVENTION

[0007] The field of the invention generally relates to bispecific antibodies, particularly T-cell redirecting bispecific antibodies (TRBAs) against cytomegalovirus (CMV).

[0008] 2. DESCRIPTION OF THE RELATED ART

[0009] A multitude of different “antibody” formats exist in the art, e.g., Fab fragments, scFv fragments, minibodies, diabodies, tribodies, tetrabodies, nanobodies, pentabodies, etc. — many of which may be bispecific or trispecific. See, e.g., Elshiaty et al. (2021).

[0010] T cell redirecting bispecific antibodies (TRBAs) are bispecific antibodies that bind to both T cells and a given target antigen. Of the many different types of bispecific antibody formats, the most common formats for TRBAs are bi-specific T cell engagers (BiTEs) and variations of heterodimeric-hybrid monoclonal antibodies.

[0011] There have been only two US FDA-approved TRBAs: Blinatumomab (anti- CD3 / anti-CD19 to treat B cell cancers, approved in 2014) and Tebentafusp (anti-CD3 linked to a T cell receptor to treat uveal cancer, approved in 2022). Outside the United States, Catumaxomab (anti-CD3 / anti-EpCAM to treat epithelial metastatic tumors in the peritoneal cavity) was previously available but was withdrawn. Blinatumomab is a BiTE (two single chain antibodies linked), Tebentafusp is BiTE-like (a single chain anti-CD3 antibody linked to a T cell receptor), and Catumaxomab was a hybrid monoclonal antibody. BiTE and hybrid monoclonal antibody formats have dominated clinical development of therapeutic bi-specific antibodies in general, due to their relative simplicity.

[0012] While there has been significant research and development of TRBAs against various cancers, there has been relatively little research and development of TRBAsagainst cytomegalovirus (CMV). In fact, to date, there are no TRBAs against CMV that have been approved for clinical use.

[0013] SUMMARY OF THE INVENTION

[0014] Provided herein are cross-over dual variable T cell directing bispecific antibodies (CODV-TRBAs), which comprise a first chain and a second chain, wherein the first chain comprises a light chain variable domain of an anti-CD3 antibody (Anti-CD3 VL) and a light chain variable domain of an anti-cytomegalovirus (CMV) antibody (Anti- CMV VL); and the second chain comprises a heavy chain variable domain of the anti- CD3 antibody (Anti-CD3 VH) and a heavy chain variable domain of the anti-CMV antibody (Anti-CMV VH), wherein the heavy chain variable domains are arranged in the opposite order as the arrangement of the light chain variable domains. In some embodiments, the Anti-CMV VL is linked to C-terminus of the Anti-CD3 VL and the Anti-CD3 VH is linked to the C-terminus of the Anti-CMV VH. In some embodiments, the Anti-CD3 VL is linked to C-terminus of the Anti-CMV VL and the Anti-CMV VH is linked to the C-terminus of the Anti-CD3 VH. In some embodiments, the CDR sequences of the anti-CD3 antibody and the anti-CMV antibody are:- Anti-CD3 CDRL1: SEQ ID NO: 1, with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL2: SEQ ID NO: 2, with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL3: SEQ ID NO: 3, with up to 1 or 2 amino acid differences;- Anti-CMV CDRL1 : SEQ ID NO: 4, with up to 1 or 2 amino acid differences;- Anti-CMV CDRL2: SEQ ID NO: 5, with up to 1 or 2 amino acid differences;- Anti-CMV CDRL3: SEQ ID NO: 6, with up to 1 or 2 amino acid differences;- Anti-CMV CDRH1 : SEQ ID NO: 7, with up to 1 or 2 amino acid differences;- Anti-CMV CDRH2: SEQ ID NO: 8, with up to 1 or 2 amino acid differences;- Anti-CMV CDRH3: SEQ ID NO: 9, with up to 1, 2, or 3 amino acid differences;- Anti-CD3 CDRH1: SEQ ID NO: 10, with up to 1 or 2 amino acid differences;- Anti-CD3 CDRH2: SEQ ID NO: 11, with up to 1, 2, or 3 amino acid differences; and- Anti-CD3 CDRH3: SEQ ID NO: 12, with up to 1 or 2 amino acid differences; wherein up to 8 of the CDR sequences have amino acid differences and / or the total number of amino acid differences in the CDR sequences is not more than 10. In some embodiments, the CDR sequences of the anti-CD3 antibody and the anti-CMV antibody are:Anti-CD3 CDRL1 : SEQ ID NO: 1;- Anti-CD3 CDRL2: SEQ ID NO: 2;- Anti-CD3 CDRL3: SEQ ID NO: 3;- Anti-CMV CDRL1 : SEQ ID NO: 4;- Anti-CMV CDRL2: SEQ ID NO: 5;- Anti-CMV CDRL3: SEQ ID NO: 6;- Anti-CMV CDRH1 : SEQ I D NO: 7;- Anti-CMV CDRH2: SEQ ID NO: 8;- Anti-CMV CDRH3: SEQ ID NO: 9;- Anti-CD3 CDRH1 : SEQ ID NO: 10;- Anti-CD3 CDRH2: SEQ ID NO: 11; and- Anti-CD3 CDRH3: SEQ ID NO: 12.In some embodiments, the sequences of framework regions of the Anti-CD3 VL and the Anti-CMV VL have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% over the combination of SEQ ID NO: 13 to SEQ ID NO: 20. In some embodiments, the sequences of framework regions of the Anti-CD3 VH and the Anti-CMV VH have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% over the combination of SEQ ID NO: 21 to SEQ ID NO: 28. In some embodiments, (a) the sequences of framework regions of the Anti-CD3 VL and the Anti-CMV VL have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% over the combination of SEQ ID NO: 13 to SEQ ID NO: 20, and (b) the sequences of framework regions of the Anti-CD3 VH and the Anti-CMV VH have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% over the combination of SEQ ID NO: 21 to SEQ ID NO: 28. In some embodiments, the Anti-CD3 VL FR1 sequence comprises or consists of SEQ ID NO: 13 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR2 sequence comprises or consists of SEQ ID NO: 14 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR3 sequence comprises or consists of SEQ I D NO: 15 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR4 sequence comprises or consists of SEQ ID NO: 16 with up to 1 or 2 amino acid differences; the Anti-CMV VL FR1 sequence comprises or consists of SEQ ID NO: 17 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR2 sequence comprises or consists of SEQ I D NO: 18 with up to 1, 2, 3, 4, or 5 amino aciddifferences; the Anti-CMV VL FR3 sequence comprises or consists of SEQ ID NO: 19 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR4 sequence comprises or consists of SEQ ID NO: 20 with up to 1 or 2 amino acid differences. In some embodiments, the Anti-CMV VH FR1 sequence comprises or consists of SEQ ID NO: 21 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR2 sequence comprises or consists of SEQ ID NO: 22 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR3 sequence comprises or consists of SEQ ID NO: 23 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR4 sequence comprises or consists of SEQ ID NO: 24 with up to 1, 2, or 3 amino acid differences; the Anti-CD3 VH FR1 sequence comprises or consists of SEQ ID NO: 25 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VH FR2 sequence comprises or consists of SEQ ID NO: 26 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VH FR3 sequence comprises or consists of SEQ ID NO: 27 with up to 1, 2, 3, 4, or 5 amino acid differences; and / or the Anti-CD3 VH FR4 sequence comprises or consists of SEQ ID NO: 28 with up to 1, 2, or 3 amino acid differences. In some embodiments, (a) the Anti-CD3 VL FR1 sequence comprises or consists of SEQ ID NO: 13 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR2 sequence comprises or consists of SEQ ID NO: 14 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR3 sequence comprises or consists of SEQ ID NO: 15 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR4 sequence comprises or consists of SEQ ID NO: 16 with up to 1 or 2 amino acid differences; the Anti-CMV VL FR1 sequence comprises or consists of SEQ ID NO: 17 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR2 sequence comprises or consists of SEQ ID NO: 18 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR3 sequence comprises or consists of SEQ ID NO: 19 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR4 sequence comprises or consists of SEQ ID NO: 20 with up to 1 or 2 amino acid differences; and (b) the Anti-CMV VH FR1 sequence comprises or consists of SEQ ID NO: 21 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR2 sequence comprises or consists of SEQ ID NO: 22 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR3 sequence comprises or consists of SEQ ID NO: 23 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR4 sequence comprises or consists of SEQ ID NO: 24 with up to 1, 2, or 3 amino acid differences; the Anti-CD3 VH FR1 sequence comprises or consists of SEQ ID NO: 25 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VH FR2 sequence comprises or consists of SEQ ID NO: 2 6 with up to 1, 2,3, 4, or 5 amino acid differences; the Anti-CD3 VH FR3 sequence comprises or consists of SEQ ID NO: 27 with up to 1, 2, 3, 4, or 5 amino acid differences; and / or the Anti-CD3 VH FR4 sequence comprises or consists of SEQ ID NO: 28 with up to 1, 2, or 3 amino acid differences. In some embodiments, the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and / or the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44. In some embodiments, the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and / or the Anti-CMV VH sequence comprises or consists of SEQ ID NO: 45. In some embodiments, the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44. In some embodiments, the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and the Anti-CMV VH sequence comprises or consists of SEQ ID NO: 45. In some embodiments, (a) the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and / or the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44 ; and (b) the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and / or the Anti- CMV VH sequence comprises or consists of SEQ I D NO: 45. In some embodiments, (a) the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44 ; and (b) the Anti-CMV VL sequence comprises or consists of SEQ I D NO: 43 and the Anti-CMV VH sequence comprises or consists of SEQ I D NO: 45. In some embodiments, the first chain comprises an antibody light chain constant region at its C-terminus. In some embodiments, the second chain comprises an antibody heavy chain constant region at its C-terminus. In some embodiments, the first chain comprises an antibody light chain constant region at its C-terminus, and the second chain comprises an antibody heavy chain constant region at its C-terminus. In some embodiments, the antibody light chain constant region comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 29. In some embodiments, the antibody heavy chain constant region comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ I D NO: 31. In some embodiments, (a) antibody light chain constant region comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 29, and (b) the antibody heavy chain constant region comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 31. In some embodiments, the amino acid sequence of the first chain is SEQ ID NO: 40. In some embodiments, the amino acid sequence of the secondchain is SEQ ID NO: 41. In some embodiments, (a) the amino acid sequence of the first chain is SEQ ID NO: 40, and (b) the amino acid sequence of the second chain is SEQ ID NO: 41. In some embodiments, the amino acid sequence of the first chain is SEQ ID NO: 4 6. In some embodiments, the amino acid sequence of the second chain is SEQ ID NO: 47. In some embodiments, (a) the amino acid sequence of the first chain is SEQ ID NO: 4 6, and (b) the amino acid sequence of the second chain is SEQ ID NO: 47. In some embodiments, the first chain and / or second chain of the CODV-TRBAs comprise a signal sequence, e.g., MDFQVQI FS FLL I SASVIMSRG ( SEQ ID NO: 35 ) or MERHWI FLSLLSVTSGVHS ( SEQ ID NO: 36 ) linked, e.g., to the N-terminus. In some embodiments, the first chain comprises SEQ ID NO: 35 at its N-terminus. In some embodiments, the second chain comprises SEQ ID NO: 36 at its N-terminus. In some embodiments, the first chain comprises SEQ ID NO: 35 at its N-terminus and the second chain comprises SEQ ID NO: 36 at its N-terminus. In some embodiments, the Anti-CD3 VL and Anti-CMV VL of the CODV-TRBAs are linked together via an amino acid linker, which comprises 4 - 10 amino acid residues, preferably 5 - 9 amino acid residues, more preferably 6 - 8 amino acid residues, most preferably 7 amino acid residues, in length. In some embodiments, the amino acid linker is TKGPS ( SEQ ID NO: 37 ) , DKTHT ( SEQ ID NO: 38 ) , or AS TKGPS ( SEQ ID NO: 39 ) . In some embodiments, the amino acid linker comprises or consists of SEQ ID NO: 37. In some embodiments, the amino acid linker is SEQ ID NO: 39. In some embodiments, the Anti-CD3 VH and Anti- CMV VH of the CODV-TRBAs are linked together via a linker comprising up to 3 amino acid residues, preferably up to 2 amino acid residues, more preferably 1 amino acid residue. In some embodiments, the Anti-CD3 VH and Anti-CMV VH of the CODV-TRBAs are linked together via a serine residue. In some embodiments, the variable regions of the CODV-TRBAs are linked to their constant regions via a linker comprising 1 - 5 amino acid residues, preferably 2 - 5 amino acid residues. In some embodiments, the amino acid linker is TKGPS ( SEQ ID NO: 37 ) , DKTHT ( SEQ ID NO: 38 ) , or Arg- Thr.

[0015] Also provided are medicaments comprising one or more CODV-TRBAs as described herein and a pharmaceutically acceptable carrier. In some embodiments, the medicaments further comprise CD8+T cells and / or one or more antivirals.

[0016] Methods of killing cells infected with a cytomegalovirus (CMV) or treating subjects for an infection by the CMV are also provided. Such methods comprise administering to the cells or the subject one or more CODV-TRBAs or a medicament asdescribed herein. In some embodiments, the methods further comprise administering CD8+T cells to the cells or the subject.

[0017] Kits comprising one or more CODV-TRBAs packaged together with CD8+T cells and / or one or more antivirals are also provided.

[0018] Both the foregoing general description and the following detailed description are exemplary and explanatory only and are intended to provide further explanation of the invention as claimed. The accompanying drawings are included to provide a further understanding of the invention and are incorporated in and constitute part of this specification, illustrate several embodiments of the invention, and together with the description explain the principles of the invention.

[0019] DESCRIPTION OF THE DRAWINGS

[0020] This invention is further understood by reference to the drawings wherein:

[0021] FIG. 1 : Schematically shows an exemplary structure of the CODV-TRBAs. Gray bars = constant regions; NL and NH = N termini of light and heavy chains, respectively; CL and CH = C termini of light and heavy chains, respectively.

[0022] FIG. 2: Clearance of CMV-infected cells when exposed to CD8+T cells in the presence of a CODV-TRBA. ARPE-19 cells were acutely infected at excess multiplicity with the GFP-expressing strain CMV-TR and then exposed to varying concentrations of the CODV-TRBA in the absence or presence of human CD8+T cells from a normal, healthy donor. Images are the inverted color pictures of cell cultures showing GFP fluorescence at 6 hours (top) and 24 hours (bottom). An additional control including the parental 12F6 antibody (recognizing only CD3) with CD8+T cells, demonstrating that non-specific CD8+T cell activation through CD3 engagement alone was not responsible for clearance of CMV-infected cells.

[0023] FIG. 3: Quantification of clearance of CMV-infected cells when exposed to CD8+T cells in the presence of the CODV-TRBA. For the experiment depicted in FIG. 2, each digital image was analyzed for the percentage of GFP fluorescent pixels.

[0024] DETAILED DESCRIPTION OF THE INVENTION

[0025] Disclosed herein are T cell redirecting bispecific antibodies (TRBAs) against cytomegalovirus (CMV) having a cross-over dual variable (CODV) antibody format, which TRBAs are hereinafter referred to as “CODV-TRBAs”. The CODV-TRBAs specifically bind a CMV and CD3 (cluster of differentiation 3) on T cells. FIG. 1 schematically shows an exemplary CODV-TRBA. As shown, the light chains of theCODV-TRBAs comprise the variable domain of the light chain of an anti-CD3 antibody (“Anti-CD3 VL”) which is linked to the variable domain of the light chain of an anti- CMV antibody (“Anti-CMV VL”) which is linked to a light chain constant region and the heavy chains comprise the variable domain of the heavy chain of the anti-CMV antibody (“Anti-CMV VH”) which is linked to the variable domain of the heavy chain of the anti-CD3 antibody (“Anti-CD3 VH”) which is linked to a heavy chain constant region. Alternatively, the light chains comprise the Anti-CMV VL which is linked to the Anti-CD3 VL which is linked to a light chain constant region and the heavy chains comprise the Anti-CD3 VH which is linked to the Anti-CMV VH which is linked to a heavy chain constant region. The linkages between two domains, e.g., the link between Anti-CMV VL and Anti-CD3 VL, or the link between the variable domain and a constant region, may be direct or indirect (e.g., via an amino acid linker).

[0026] In some embodiments, the sequences of CDRs of the Anti-CD3 VL, Anti-CMV VL, Anti-CMV VH, and Anti-CD3 VH are:- Anti-CD3 CDRLL RASSSVSYMH ( SEQ ID NO: 1 ) , with up to 1 or 2 amino acid differences- Anti-CD3 CDRL2: ATSNLAS ( SEQ ID NO: 2 ) , with up to 1 or 2 amino acid differences- Anti-CD3 CDRL3: QQWSSNPPT ( SEQ ID NO: 3 ) , with up to 1 or 2 amino acid differences- Anti-CMV CDRL1 : QSVSND ( SEQ ID NO: 4 ) , with up to 1 or 2 amino acid differences- Anti-CMV CDRL2: YAS ( SEQ ID NO: 5 ) , with up to 1 or 2 amino acid differences- Anti-CMV CDRL3: QQDYSSPWT ( SEQ ID NO: 6 ) , with up to 1 or 2 amino acid differences- Anti-CMV CDRH1: GYT FT I YG ( SEQ ID NO: 7 ) , with up to 1 or 2 amino acid differences- Anti-CMV CDRH2: INTYTGEP ( SEQ ID NO: 8 ) , with up to 1 or 2 amino acid differences- Anti-CMV CDRH3: ARKGYYGSSGYFDY ( SEQ ID NO: 9 ) , with up to 1, 2, or 3 amino acid differences- Anti-CD3 CDRH1: SYTMH ( SEQ ID NO: 10 ) , with up to 1 or 2 amino acid differences- Anti-CD3 CDRH2: YINPSSGYTKYNQKFKD ( SEQ ID NO: 11 ) , with up to 1, 2, or 3 amino acid differences- Anti-CD3 CDRH3: WQDYDVYFDY ( SEQ ID NO: 12 ) , with up to 1 or 2 amino acid differences, wherein up to 8, up to 7, up to 6, up to 5, up to 4, up to 3, up to 2, 1, or none of the CDR sequences have amino acid differences and / or the total number of amino acid differences in the CDR sequences is not more than 10, not more than 9, not more than 8, not more than 7, not more than 6, not more than 5, not more than 4, not more than 3, not more than 2, 1, or 0. In some embodiments, the sequences of the CDRs of the CODV-TRBAs comprise or consist of SEQ ID NO: 1 to SEQ ID NO: 12.

[0027] As exemplified herein, the sequences of framework regions of Anti-CD3 VL and Anti-CMV VL are:- Anti-CD3 VLFR1: QIVLSQSPAILSASPGEKVTMTC (SEQ ID NO: 13)- Anti-CD3 VLFR2: WYQQKPGSSPKPWIY (SEQ ID NO: 14)- Anti-CD3 VLFR3: GVPARFSGSGSGTSYSLTISRVEAEDAATYYC (SEQ ID NO:15)- Anti-CD3 VLFR4: FGGGTKLEIK (SEQ ID NO: 16)- Anti-CMV VL FR1 : DIVMTQTPKFLLVSAGDRVTITCKAS (SEQ ID NO: 17)- Anti-CMV VLFR2: VSWYQQKPGQSPKLLIY (SEQ IDNO: 18)- Anti-CMV VLFR3: NRYTGVPDRFTGSGYGTDFTFTISTVQAEDLAVYFC (SEQ ID NO: 19)- Anti-CMV VL FR4: FGGGTKLEIK (SEQ ID NO: 20)

[0028] As exemplified herein, the sequences of framework regions of Anti-CMV VH and Anti-CD3 VH are:- Anti-CMV VHFR1: QIQLVQSGPELKKPGETVKISCKAS (SEQ IDNO: 21)- Anti-CMV VHFR2: MNWVKQAPGKGLKWMGW (SEQ ID NO: 22)- Anti-CMV VHFR3: TYADDFRGRFAFSLETSASTAYLQINNLKNEDTATYFC (SEQ ID NO: 23)- Anti-CMV VHFR4: WGQGTTLTVSS (SEQ IDNO: 24)- Anti-CD3 VHFR1: QVQLQQSGAELARPGASVKMSCKASGYTFT (SEQ ID NO: 25)- Anti-CD3 VHFR2: WVKQRPGQGLEWIG (SEQ ID NO: 26)- Anti-CD3 VHFR3: KATLTADKSSSTAYMQLSSLTSEDSAVYYCAR (SEQ ID NO:27)- Anti-CD3 VHFR4: WGQGTTLTVSS (SEQ ID NO: 28)

[0029] Suitable constant regions include those known in the art. In some embodiments, the light chain constant region is the following Light Chain Kappa Constant Region: RTVAAPSVFI FPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDST YSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC ( SEQ ID NO: 29 )

[0030] In some embodiments, the light chain constant region is the following LightChain Lambda Constant Region:GQPKANPTVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADGSPVKAGVETTKPSKQSNNK YAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS ( SEQ ID NO: 30 )

[0031] In some embodiments, the heavy chain constant region is the following HeavyChain IgG3 Constant Region:ASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYTCNVNHKPSNTKVDKRVELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCP EPKSCDTPPPCPRCPEPKSCDTPPPCPRCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVD VSHEDPEVQFKWYVDGVEVHNAKTKLREEQYNSTFRVVSVLTVLHQDWLNGKEYKCKVSNKALPA PIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYNTTP PMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNHYTQKSLSLSPGK ( SEQ ID NO: 31 )

[0032] In some embodiments, the heavy chain constant region is the following HeavyChain IgGl Constant Region:ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKP KDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQD WLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDI AVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPGK ( SEQ ID NO: 32 )

[0033] In some embodiments, the heavy chain constant region is the following HeavyChain IgG2 Constant Region:ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSNFGTQTYTCNVDHKPSNTKVDKTVERKCCVECPPCPAPPVAGPSVFLFPPKPKDTL MISRTPEVTCVVVDVSHEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTFRVVSVLTVVHQDWLNG KEYKCKVSNKGLPAPIEKTISKTKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDISVEW ESNGQPENNYKTTPPMLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPG K ( SEQ ID NO: 33 )

[0034] In some embodiments, the heavy chain constant region is the following HeavyChain IgG4 Constant Region:ASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSL SSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPSCPAPEFLGGPSVFLFPPKPKDT LMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLN GKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSL GK ( SEQ ID NO: 34 )

[0035] In some embodiments, the sequences of the non-CDR regions (z.e., framework regions and, if present, the constant region) of the light chain of CODV-TRBAs have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, atleast 99%, or 100% over the combination of SEQ ID NO: 13 to SEQ ID NO: 20 or over the combination of SEQ ID NO: 13 to SEQ ID NO: 29. For example, where the collective sequence identity is at least 95%, the first framework region (FR1) of the Anti-CD3 VL may have less than 95% sequence identity to SEQ ID NO: 13 so long as the number of identical residues over the remaining framework and constant regions raise the sum total of identical residues to at least 95%. In some embodiments, the Anti-CD3 VL FR1 sequence comprises or consists of SEQ ID NO: 13 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR2 sequence comprises or consists of SEQ ID NO: 14 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR3 sequence comprises or consists of SEQ ID NO: 15 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR4 sequence comprises or consists of SEQ ID NO: 16 with up to 1 or 2 amino acid differences; the Anti-CMV VL FR1 sequence comprises or consists of SEQ ID NO: 17 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR2 sequence comprises or consists of SEQ ID NO: 18 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR3 sequence comprises or consists of SEQ ID NO: 19 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR4 sequence comprises or consists of SEQ ID NO: 20 with up to 1 or 2 amino acid differences; and / or the light chain constant region sequence comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 29.

[0036] In some embodiments, the sequences of the non-CDR regions (z.e., framework regions, if present, the constant region) of the heavy chain of CODV-TRBAs have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% over the combination of SEQ ID NO: 21 to SEQ ID NO: 28 or over the combination of SEQ ID NO: 21 to SEQ ID NO: 28, and SEQ ID NO: 31. For example, where the collective sequence identity is at least 95%, the third framework region (FR3) of the Anti-CD3 VH may have less than 95% sequence identity to SEQ ID NO: 27 so long as the number of identical residues over the remaining framework and constant regions raise the sum total of identical residues to at least 95%. In some embodiments, the Anti- CMV VH FR1 sequence comprises or consists of SEQ ID NO: 21 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR2 sequence comprises or consists of SEQ ID NO: 22 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR3 sequence comprises or consists of SEQ ID NO: 23 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR4 sequence comprises or consists of SEQ ID NO: 24with up to 1, 2, or 3 amino acid differences; the Anti-CD3 VH FR1 sequence comprises or consists of SEQ ID NO: 25 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti- CD3 VH FR2 sequence comprises or consists of SEQ ID NO: 2 6 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VH FR3 sequence comprises or consists of SEQ ID NO: 27 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VH FR4 sequence comprises or consists of SEQ ID NO: 28 with up to 1, 2, or 3 amino acid differences; and / or the heavy chain constant region sequence comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 31.

[0037] In some embodiments, the Anti-CD3 VL FR1 sequence comprises or consists of SEQ ID NO: 13; the Anti-CD3 VL FR2 sequence comprises or consists of SEQ ID NO: 14 ; the Anti-CD3 VL FR3 sequence comprises or consists of SEQ ID NO: 15 ; the Anti-CD3 VL FR4 sequence comprises or consists of SEQ ID NO: 16; the Anti-CMV VL FR1 sequence comprises or consists of SEQ ID NO: 17; the Anti-CMV VL FR2 sequence comprises or consists of SEQ ID NO: 18; the Anti-CMV VL FR3 sequence comprises or consists of SEQ ID NO: 19; the Anti-CMV VL FR4 sequence comprises or consists of SEQ ID NO: 20; the light chain constant region sequence comprises or consists of SEQ ID NO: 2 9; the Anti-CMV VH FR1 sequence comprises or consists of SEQ ID NO: 21; the Anti-CMV VH FR2 sequence comprises or consists of SEQ ID NO: 22; the Anti-CMV VH FR3 sequence comprises or consists of SEQ ID NO: 23; the Anti-CMV VH FR4 sequence comprises or consists of SEQ ID NO: 24; the Anti-CD3 VH FR1 sequence comprises or consists of SEQ ID NO: 25; the Anti-CD3 VH FR2 sequence comprises or consists of SEQ ID NO: 26; the Anti-CD3 VH FR3 sequence comprises or consists of SEQ ID NO: 27 ; the Anti-CD3 VH FR4 sequence comprises or consists of SEQ ID NO: 28 ; and the heavy chain constant region sequence comprises or consists of SEQ ID NO: 31.

[0038] In some embodiments, the light chain and / or heavy chain of the CODV-TRBAs comprise a signal sequence, e.g., MDFQVQI FS FLL I SASVIMSRG ( SEQ ID NO: 35 ) or MERHWI FLSLLSVTSGVHS ( SEQ ID NO: 36 ) linked, e.g., to the N-terminus.

[0039] In some embodiments, the Anti-CD3 VL and Anti-CMV VL of the CODV- TRBAs are linked together via an amino acid linker, which comprises 4 - 10 amino acid residues, preferably 5 - 9 amino acid residues, more preferably 6 - 8 amino acid residues, most preferably 7 amino acid residues, in length. Suitable linkers include those known and used in the art to link antibody variable domains. See, e.g., Xu et al. (2017). In some embodiments, the amino acid linker is TKGPS ( SEQ ID NO: 37 ) , DKTHT ( SEQID NO: 38 ) , or AS TKGPS ( SEQ ID NO: 39 ) . In some embodiments, the amino acid linker comprises or consists of SEQ ID NO: 37. In some embodiments, the amino acid linker is SEQ ID NO: 39.

[0040] In some embodiments, the Anti-CD3 VH and Anti-CMV VH of the CODV- TRBAs are linked together via a linker comprising up to 3 amino acid residues, preferably up to 2 amino acid residues, more preferably 1 amino acid residue. In some embodiments, the Anti-CD3 VH and Anti-CMV VH of the CODV-TRBAs are linked together via a serine residue.

[0041] In some embodiments, the variable regions of the CODV-TRBAs are linked to their constant regions via a linker comprising 1 - 5 amino acid residues, preferably 2 - 5 amino acid residues. Suitable linkers include those known and used in the art to link antibody variable domains. See, e.g., Xu et al. (2017). In some embodiments, the amino acid linker is TKGPS ( SEQ ID NO: 37 ) , DKTHT ( SEQ ID NO: 38 ) , or Arg- Thr.

[0042] In some embodiments, the light chains of the CODV-TRBAs comprise or consist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 40, and the heavy chains of the CODV-TRBAs comprise or consist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 41, wherein the sequences of the CDRs are:- Anti-CD3 CDRL1 : RAS S SVSYMH ( SEQ ID NO: 1 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL2: ATSNLAS ( SEQ ID NO: 2 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL3: QQWS SNPPT ( SEQ ID NO: 3 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRL1 : QSVSND ( SEQ ID NO: 4 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRL2: YAS ( SEQ ID NO: 5 ) , with up to 1 or 2 amino acid differences,- Anti-CMV CDRL3: QQDYS S PWT ( SEQ ID NO: 6 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH1: GYT FT I YG ( SEQ ID NO: 7 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH2: INTYTGEP ( SEQ ID NO: 8 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH3: ARKGYYGSSGYFDY ( SEQ ID NO: 9 ) , with up to 1, 2, or 3 amino acid differences;- Anti-CD3 CDRH1: SYTMH ( SEQ ID NO: 10 ), with up to 1 or 2 amino acid differences;- Anti-CD3 CDRH2: YINPSSGYTKYNQKFKD ( SEQ ID NO: 11 ), with up to 1, 2, or 3 amino acid differences; and- Anti-CD3 CDRH3: WQDYDVYFDY ( SEQ ID NO: 12 ) , with up to 1 or 2 amino acid differences; and wherein up to 8, up to 7, up to 6, up to 5, up to 4, up to 3, up to 2, 1, or none of the CDR sequences have amino acid differences and / or the total number of amino acid differences in the CDR sequences is not more than 10, not more than 9, not more than 8, not more than 7, not more than 6, not more than 5, not more than 4, not more than 3, not more than 2, 1, or 0.

[0043] In some embodiments, the light chains of the CODV-TRBAs comprise or consist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 40, and the heavy chains of the CODV-TRBAs comprise or consist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 41, wherein the sequences of the CDRs are:- Anti-CD3 CDRL1: RASSSVSYMH (SEQ IDNO: 1);- Anti-CD3 CDRL2: ATSNLAS (SEQ ID NO: 2 ) ;- Anti-CD3 CDRL3: QQWSSNPPT (SEQ IDNO: 3);- Anti-CMV CDRL1 : QSVSND ( SEQ ID NO: 4 ) ;- Anti-CMV CDRL2: YAS ( SEQ ID NO: 5 ) ;- Anti-CMV CDRL3: QQDYSSPWT (SEQ ID NO: 6) ;- Anti-CMV CDRH1: GYT FT I YG (SEQ IDNO: 7);- Anti-CMV CDRH2: INTYTGEP (SEQ ID NO: 8 ) ;- Anti-CMV CDRH3: ARKGYYGSSGYFDY (SEQ ID NO: 9);- Anti-CD3 CDRH1: SYTMH (SEQ ID NO: 10) ;- Anti-CD3 CDRH2: YINPSSGYTKYNQKFKD (SEQ ID NO: 11) ; and- Anti-CD3 CDRH3: WQDYDVYFDY (SEQ ID NO: 12) .

[0044] In some embodiments, the light chains of the CODV-TRBAs comprise or consist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 46, and the heavy chains of the CODV-TRBAs comprise orconsist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 47, wherein the sequences of the CDRs are:- Anti-CD3 CDRL1: RASSSVSYMH ( SEQ ID NO: 1 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL2: ATSNLAS ( SEQ ID NO: 2 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL3: QQWS SNPPT ( SEQ ID NO: 3 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRL1 : QSVSND ( SEQ ID NO: 4 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRL2: YAS ( SEQ ID NO: 5 ) , with up to 1 or 2 amino acid differences,- Anti-CMV CDRL3: QQDYS S PWT ( SEQ ID NO: 6 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH1 : GYT FT I YG ( SEQ ID NO: 7 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH2: INTYTGEP ( SEQ ID NO: 8 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH3: ARKGYYGS SGYFDY ( SEQ ID NO: 9 ) , with up to 1, 2, or 3 amino acid differences;- Anti-CD3 CDRH1 : SYTMH ( SEQ ID NO: 10 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRH2: YINPS SGYTKYNQKFKD ( SEQ ID NO: 11 ) , with up to 1, 2, or 3 amino acid differences; and- Anti-CD3 CDRH3: WQDYDVYFDY ( SEQ ID NO: 12 ) , with up to 1 or 2 amino acid differences; and wherein up to 8, up to 7, up to 6, up to 5, up to 4, up to 3, up to 2, 1, or none of the CDR sequences have amino acid differences and / or the total number of amino acid differences in the CDR sequences is not more than 10, not more than 9, not more than 8, not more than 7, not more than 6, not more than 5, not more than 4, not more than 3, not more than 2, 1, or 0.

[0045] In some embodiments, the light chains of the CODV-TRBAs comprise or consist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 4 6, and the heavy chains of the CODV-TRBAs comprise orconsist of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 47, wherein the sequences of the CDRs are:- Anti-CD3 CDRL1: RASSSVSYMH ( SEQ ID NO: 1 ) ;- Anti-CD3 CDRL2: ATSNLAS ( SEQ ID NO: 2 ) ;- Anti-CD3 CDRL3: QQWS SNPPT ( SEQ ID NO: 3 ) ;- Anti-CMV CDRL1 : QSVSND ( SEQ ID NO: 4 ) ;- Anti-CMV CDRL2: YAS ( SEQ ID NO: 5 ) ;- Anti-CMV CDRL3: QQDYS S PWT ( SEQ ID NO: 6 ) ;- Anti-CMV CDRH1 : GYT FT I YG ( SEQ ID NO: 7 ) ;- Anti-CMV CDRH2: INTYTGEP ( SEQ ID NO: 8 ) ;- Anti-CMV CDRH3: ARKGYYGS SGYFDY ( SEQ ID NO: 9 ) ;- Anti-CD3 CDRH1 : SYTMH ( SEQ ID NO: 10 ) ;- Anti-CD3 CDRH2: YINPS SGYTKYNQKFKD ( SEQ ID NO: 11 ) ; and- Anti-CD3 CDRH3: WQDYDVYFDY ( SEQ ID NO: 12 ) .

[0046] In some embodiments, the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and / or the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44.

[0047] In some embodiments, the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and / or the Anti-CMV VH sequence comprises or consists of SEQ ID NO: 45.

[0048] In some embodiments, the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44.

[0049] In some embodiments, the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and the Anti-CMV VH sequence comprises or consists of SEQ ID NO: 45.

[0050] In some embodiments, (a) the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and / or the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44 ; and (b) the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and / or the Anti-CMV VH sequence comprises or consists of SEQ ID NO: 45.

[0051] In some embodiments, (a) the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44; and (b) the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and the Anti-CMV VH sequence comprises or consists of SEQ ID NO: 45.

[0052] The mechanism in which TRBAs exert their therapeutic effect is by cross-linking CD8+killer T cell (CTL) to a target cell. Generally, a TRBA cross-links the cells by binding both CD3 expressed on CTLs and an antigen expressed on the surface of thetarget cell. This cross-linking activates the CTL to kill the target cell. TRBAs having an antibody heavy chain Fc domain can also recruit complement or NK cells to kill the target cell. Thus, the CODV-TRBAs preferably comprise an antibody heavy chain Fc domain.

[0053] As shown in FIG. 2 and FIG. 3, when CMV-infected cells were treated with either (a) CODV-TRBA alone, or (b) CD8+T cells alone, there was no reduction of CMV infection, but the combination of CODV-TRBA and CD8+T cells resulted in rapid reduction in infected cells. These results indicate that the CODV-TRBAs cross-link CD8+T cells and CMV-infected cells, which thereby leads to the CTL clearance of the CMV-infected cells. Particularly, in the presence of CD8+T cells the CODV-TRBA resulted in a marked clearance of the infected cells at 6 hours and nearly complete clearance of the infected cells by 24 hours.

[0054] Therefore, one or more CODV-TRBAs may be used to kill cells infected with CMV or treat a CMV infection in a subject. Such methods comprise administering the one or more CODV-TRBAs to the cells or the subject in the presence of CD8+T cells. In some embodiments, the CD8+T cells are co-administered with the one or more CODV-TRBAs.

[0055] As used herein, a “CMV infection” refers to an infection by a CMV (ie., a virus belonging to the Cytomegalovirus genus). Viruses belonging to the Cytomegalovirus genus include Aotine betaherpesvirus 7, Cebine betaherpesvirus 7, Cercopithecine betaherpesvirus 5, Human betaherpesvirus 5 (human CMV), Macacine betaherpesvirus3, Macacine betaherpesvirus 8, Mandrilline betaherpesvirus 7, Panine betaherpesvirus 2, Papiine betaherpesvirus 3, Papiine betaherpesvirus 4. and Saimiriine betaherpesvirus4. In preferred embodiments, the CMV is human CMV.

[0056] CODV-TRBAs may be made using methods known in the art including chemical synthesis, biosynthesis or in vitro synthesis using recombinant DNA methods, and solid phase synthesis. See, e.g, Kelly & Winkler (1990) Genetic Engineering Principles and Methods, vol. 12, J.K. Setlow ed., Plenum Press, NY, pp. 1-19; Merrifield (1964) J Amer Chem Soc 85:2149; Houghten (1985) PNAS USA 82:5131-5135; and Stewart & Young (1984) Solid Phase Peptide Synthesis, 2 ed. Pierce, Rockford, IL, which are herein incorporated by reference. CODV-TRBAs may be purified using protein purification techniques known in the art such as reverse phase high-performance liquid chromatography (HPLC), ion-exchange or immunoaffinity chromatography, filtration or size exclusion, or electrophoresis. See, e.g, Olsnes and Pihl (1973) Biochem 12(16):3121-3126; and Scopes (1982) Protein Purification, Springer-Verlag, NY, whichare herein incorporated by reference. Alternatively, CODV-TRBAs may be made by recombinant DNA techniques known in the art. Thus, polynucleotides that encode CODV-TRBAs are contemplated herein. In some embodiments, the polypeptides and polynucleotides are isolated.

[0057] As used herein, an “isolated” compound refers to a compound that is isolated from its native environment. For example, an isolated polynucleotide is a one which does not have the bases normally flanking the 5’ end and / or the 3’ end of the polynucleotide as it is found in nature. As another example, an isolated polypeptide is a one which does not have its native amino acids, which correspond to the full-length polypeptide, flanking the N-terminus, C-terminus, or both. For example, an isolated fragment of CODV-TRBA refers to an isolated polypeptide that consists of only a portion of the CODV-TRBA or comprises some, but not all, of the amino acid residues CODV-TRBA and non-native amino acids, z.e., amino acids that are different from the amino acids found at the corresponding positions of CODV-TRBA, at its N-terminus, C- terminus, or both. In some embodiments, isolated polynucleotides and polypeptides are made “by the hand of man”, e.g., using synthetic and / or recombinant techniques.

[0058] In some embodiments, the CODV-TRBAs are substantially purified. As used herein, a “substantially purified” compound refers to a compound that is removed from its natural environment and / or is at least about 60% free, preferably about 75% free, and more preferably about 90% free, and most preferably about 95-100% free from other macromolecular components or compounds with which the compound is associated with in nature or from its synthesis.

[0059] The CODV-TRBAs specifically bind CD3 and CMV, preferably human CMV. As used herein, a compound (e.g., antibody) “specifically binds” a given target (e.g., ligand or epitope) if it reacts or associates more frequently, more rapidly, with greater duration, and / or with greater binding affinity with the given target than it does with a given alternative, and / or indiscriminate binding that gives rise to non-specific binding and / or background binding. As used herein, “non-specific binding” and “background binding” refer to an interaction that is not dependent on the presence of a specific structure (e.g., a given epitope).

[0060] As used herein, “binding affinity” refers to the propensity of a compound to associate with (or alternatively dissociate from) a given target and may be expressed in terms of its dissociation constant, Kd. In some embodiments, the antibodies have a Kd of 10'5or less, 10'6or less, preferably 10'7or less, more preferably 10'8or less, even more preferably 10'9or less, and most preferably 10'10or less, to their given target.Binding affinity can be determined using methods in the art, such as equilibrium dialysis, equilibrium binding, gel filtration, immunoassays, surface plasmon resonance, and spectroscopy using experimental conditions that exemplify the conditions under which the compound and the given target may come into contact and / or interact. Dissociation constants may be used determine the binding affinity of a compound for a given target relative to a specified alternative, e.g., bovine serum albumin. Alternatively, methods in the art, e.g., immunoassays, in vivo or in vitro assays for functional activity, etc., may be used to determine the binding affinity of the compound for the given target relative to the specified alternative. Thus, in some embodiments, the binding affinity of the CODV- TRBAs for CD3 and a CMV is at least 1-fold or more, preferably at least 5-fold or more, more preferably at least 10-fold or more, and most preferably at least 100-fold or more than its binding affinity for the specified alternative.

[0061] As used herein, “co-administration” refers to the administration of at least two different agents, i.e., first agent (e.g., one or more CODV-TRBAs) and a second agent (e.g., one or more antivirals or CD8+T cells) to a subject. In some embodiments, the coadministration is concurrent. In embodiments involving concurrent co-administration, the agents may be administered as a single composition, e.g., an admixture, or as two separate compositions. In some embodiments, the first agent is administered before and / or after the administration of the second agent. Where the co-administration is sequential, the administration of the first and second agents may be separated by a period of time, e.g., minutes, hours, or days. Those of skill in the art understand that the formulations and / or routes of administration of the various agents or therapies used may vary. The appropriate dosage for co-administration can be readily determined by one skilled in the art. In some embodiments, when two or more agents are co-administered, the respective agents are administered at lower dosages than appropriate for their administration alone.

[0062] Kits

[0063] In some embodiments, the present invention provides kits comprising one or more CODV-TRBAs, optionally in a composition or in combination with one or more supplementary agents, packaged together with one or more reagents or drug delivery devices for preventing, inhibiting, reducing, or treating a CMV infection in a subject. In some embodiments, the kits comprise the one or more CODV-TRBAs, optionally in one or more unit dosage forms, packaged together as a pack and / or in drug delivery device, e.g., a pre-filled syringe.

[0064] In some embodiments, the kits include a carrier, package, or container that may be compartmentalized to receive one or more containers, such as vials, tubes, and the like. In some embodiments, the kits optionally include an identifying description or label or instructions relating to its use. In some embodiments, the kits include information prescribed by a governmental agency that regulates the manufacture, use, or sale of compounds and compositions as contemplated herein.

[0065] Compositions

[0066] Compositions, including pharmaceutical compositions, comprising, consisting essentially of, or consisting of one or more CODV-TRBAs are contemplated herein. The term “pharmaceutical composition” refers to a composition suitable for pharmaceutical use in a subject. A composition generally comprises an effective amount of an active agent and a diluent and / or carrier. A pharmaceutical composition generally comprises a therapeutically effective amount of an active agent and a pharmaceutically acceptable carrier. In addition to the one or more CODV-TRBAs, pharmaceutical compositions may include one or more supplementary agents. Examples of suitable supplementary agents include antivirals such as Cidofovir, Foscarnet (Phosphonoformic acid), Ganciclovir, Letermovir, Valganciclovir, and the like.

[0067] As used herein, an “effective amount” refers to a dosage or amount sufficient to produce a desired result. The desired result may comprise an objective or subjective change as compared to a control in, for example, in vitro assays, and other laboratory experiments. As used herein, a “therapeutically effective amount” refers to an amount that may be used to treat, prevent, or inhibit a given disease or condition in a subject as compared to a control, such as a placebo. Again, the skilled artisan will appreciate that certain factors may influence the amount required to effectively treat a subject, including the degree of the condition or symptom to be treated, previous treatments, the general health and age of the subject, and the like. Nevertheless, effective amounts and therapeutically effective amounts may be readily determined by methods in the art.

[0068] The one or more CODV-TRBAs may be administered, preferably in the form of pharmaceutical compositions, to a subject. Preferably the subject is mammalian, more preferably, the subject is human. Preferred pharmaceutical compositions are those comprising at least one CODV-TRBA in a therapeutically effective amount and a pharmaceutically acceptable vehicle. It should be noted that treatment of a subject with a therapeutically effective amount may be administered as a single dose or as a series of several doses. The dosages used for treatment may increase or decrease over the courseof a given treatment. Optimal dosages for a given set of conditions may be ascertained by those skilled in the art using dosage-determination tests and / or diagnostic assays in the art. Dosage-determination tests and / or diagnostic assays may be used to monitor and adjust dosages during the course of treatment.

[0069] Pharmaceutical compositions may be formulated for the intended route of delivery and administered to subjects accordingly using methods in the art. It will be appreciated that the preferred route of administration and pharmaceutical formulation will vary with the condition and age of the subject, the nature of the condition to be treated, the therapeutic effect desired, and the particular CODV-TRBA used.

[0070] Pharmaceutical compositions may include one or more of the following: a pharmaceutically acceptable vehicle, pH buffered solutions, preservatives, wetting agents, emulsifying agents, and dispersing agents, liposomal formulations, nanoparticles, dispersions, suspensions, or emulsions, as well as sterile powders for reconstitution into sterile injectable solutions or dispersions. The compositions and formulations may be optimized for increased stability and efficacy using methods in the art. See, e.g., Carra et al., (2007) Vaccine 25:4149-4158.

[0071] As used herein, a “pharmaceutically acceptable vehicle” or “pharmaceutically acceptable carrier” are used interchangeably and refer to solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying agents, and the like, that are compatible with pharmaceutical administration and comply with the applicable standards and regulations, e.g., the pharmacopeial standards set forth in the United States Pharmacopeia and the National Formulary (USP-NF) book, for pharmaceutical administration. Thus, for example, unsterile water is excluded as a pharmaceutically acceptable carrier for, at least, intravenous administration. Pharmaceutically acceptable vehicles include those known in the art. See, e.g., Remington: The Science and Practice of Pharmacy 20th ed (2000) Lippincott Williams & Wilkins, Baltimore, MD.

[0072] The pharmaceutical compositions may be provided in dosage unit forms. As used herein, a “dosage unit form” refers to physically discrete units suited as unitary dosages for the subject to be treated; each unit containing a predetermined quantity of the one or more CODV-TRBA calculated to produce the desired therapeutic effect in association with the required pharmaceutically acceptable carrier. The specification for the dosage unit forms of the invention are dictated by and directly dependent on the unique characteristics of the given CODV-TRBA and desired therapeutic effect to beachieved, and the limitations inherent in the art of compounding such an active compound for the treatment of individuals.

[0073] Toxicity and therapeutic efficacy of CODV-TRBAs according to the instant invention and compositions thereof can be determined using cell cultures and / or experimental animals and pharmaceutical procedures in the art. While CODV-TRBAs that result in toxic side-effects may be used, care should be taken to design a delivery system that targets such compounds to the site of treatment to minimize potential damage to uninfected cells and, thereby, reduce side-effects.

[0074] The data obtained from the cell culture assays and animal studies can be used in formulating a range of dosages for use in humans. Preferred dosages provide a range of circulating concentrations that include the EDso with little or no toxicity. The dosage may vary depending upon the dosage form employed and the route of administration utilized. Therapeutically effective amounts and dosages of one or more CODV-TRBAs can be estimated initially from cell culture assays. Additionally, a dosage suitable for a given subject can be determined by an attending physician or qualified medical practitioner, based on various clinical factors.

[0075] The following examples are intended to illustrate but not to limit the invention.

[0076] EXAMPLES

[0077] Production of Exemplary CODV-TRBA

[0078] An exemplary CODV-TRBA was constructed using recombinant techniques in the art. See, e.g., Steinmetz et al. (2016). The variable chain genes of the known anti- CMV antibody “21E9” and the known anti-CD3 antibody “12F6” were used to create the following light and heavy chains:

[0079] Light Chain:MDFQVQI FS FLL I SASVIMSRGQIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQQ KPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPT FGGGTKLEIKAS T KG P S D IVMTQTPKFLLVSAGDRVT TCKASQSVSNDVSWYQQKPGQSPKLLIYYASNRYTGVPDRFTGSGYGTDFTFTISTVQAEDLAVYFCQQDYSSPWTFGGG TKLEIKTKGPSRTVAAPSVFI FPPSDEQLKSGTASWCLLNNFYPREAKVQWKVDNALQ SGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC ( SEQ ID NO: 40 )

[0080] Heavy Chain:MERHWI FLSLLSVTS G VH S OIQLVOSGPELKKPGETVKISCKASGYTFTIYGMNWVKQA PGKGLKWMGWINTYTGEPTYADDFRGRFAFSLETSASTAYLQINNLKNEDTATYFCARK GYYGSSGYFDYWGOGTTLTVSSSQVQLQQSGAELARPGASVKMSCKASGYTFTSYTMHW VKQRPGQGLEWIGYINPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTSEDSAVYY CARWQDYDVYFDYWGQGTTLTVSSRTASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYF PEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSWTVPSSSLGTQTYTCNVNHKPSNT KVDKRVELKTPLGDTTHTCPRCPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPEPKSCD TPPPCPRCPAPELLGGPSVFLFPPKPKDTLMI SRTPEVTCVWDVSHEDPEVQFKWYVD GVEVHNAKTKLREEQYNSTFRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT I SKA KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYNTTPPML DSDGS FFLYSKLTVDKSRWQQGNI FSCSVMHEALHNHYTQKSLSLSPGK ( SEQ ID NO: 41 )

[0081] In the above light chain ( SEQ ID NO: 40 ) and heavy chain ( SEQ ID NO: 41 ) , the underlined normal font indicates the Anti-CD3 VL sequence ( SEQ ID NO: 42 ) , underlined bold font indicates the Anti-CMV VL sequence ( SEQ ID NO: 43 ) , double underlined normal font indicates the Anti-CD3 VH sequence ( SEQ ID NO: 44 ) , and the double underlined bold font indicate the Anti-CMV VH sequence ( SEQ ID NO: 45 ) . Using methods in the art, expression vectors encoding the light chain and expression vectors encoding the heavy chain were co-transfected in to Human embryonic kidney (HEK) 293T cells, which were then cultured under conditions to produce the CODV- TRBA.

[0082] As indicated above, in alternative embodiments, the light chains comprise the Anti-CMV VL which is linked to the Anti-CD3 VL which is linked to a light chain constant region and the heavy chains comprise the Anti-CD3 VH which is linked to the Anti-CMV VH which is linked to a heavy chain constant region. The following light and heavy chains exemplify such an alternative embodiment:

[0083] Alternative Light Chain:MD EQ VQ I FS ELL I S S VIMS RGDIVMTQTPKFLLVSAGDRVTITCKASQSVSNDVSWYQQKPGQS PKLLIYYASNRYTGVPDRFTGSGYGTDFTFTISTVQAEDLAVYFCQQDYSSPWTFGGGTKLEIKA STKGPSQIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQQKPGSSPKPWIYATSNLASGVPA RFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPPTFGGGTKLEIKT KG P S RT VA P S V F I F P P SDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKAD YEKHKVYACEVTHQGLSSPVTKSFNRGEC ( SEQ ID NO: 46 )

[0084] Alternative Heavy Chain:MERHWI FLSLLSVTSGVH SQVQLQQSGAELARPGASVKMSCKASGYTFTSYTMHWVKQRPGQGLE WIGYINPSSGYTKYNQKFKDKATLTADKSSSTAYMQLSSLTSEDSAVYYCARWQDYDVYFDYWGQ GTTLTVSSSQIQLVQSGPELKKPGETVKISCKASGYTFTIYGMNWVKQAPGKGLKWMGWINTYTG EPTYADDFRGRFAFSLETSASTAYLQINNLKNEDTATYFCARKGYYGSSGYFDYWGQGTTLTVSS RTASTKGPSVFPLAPCSRSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLY SLSSVVTVPSSSLGTQTYTCNVNHKPSNTKVDKRVELKTPLGDTTHTCPRCPEPKSCDTPPPCPR CPEPKSCDTPPPCPRCPEPKSCDTPPPCPRCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVV VDVSHEDPEVQFKWYVDGVEVHNAKTKLREEQYNSTFRVVSVLTVLHQDWLNGKEYKCKVSNKAL PAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYNT TPPMLDSDGSFFLYSKLTVDKSRWQQGNIFSCSVMHEALHNHYTQKSLSLSPGK ( SEQ ID NO: 47 )

[0085] In the above light chain ( SEQ ID NO: 46 ) and heavy chain ( SEQ ID NO: 47 ) , the underlined normal font indicates the Anti-CD3 VL sequence ( SEQ ID NO: 42 ) , underlined bold font indicates the Anti-CMV VL sequence ( SEQ ID NO: 43 ) , double underlined normal font indicates the Anti-CD3 VH sequence ( SEQ ID NO: 44 ) , and the double underlined bold font indicate the Anti-CMV VH sequence ( SEQ ID NO: 45 ) .

[0086] CODV-TRBAs Direct T Cells to Destroy CMV-infected Target Cells

[0087] The functional activity of the CODV-TRBA was tested by contacting CMV- infected cells with CD8+T cells (from a healthy donor) (a) alone, or (b) in combination with the CODV-TRBA. The CMV-infected cells were infected with a strain of CMV that expresses green fluorescent protein was used, whereby the amount of fluorescence is indicative of the amount of CMV infection.

[0088] The functional activity of the CODV-TRBA was tested by contacting CMV- infected cells with CD8+T cells in combination with various concentrations of the CODV-TRBA or with a control anti-CD3 antibody (that was the parental source of the anti-CD3 portion of the CODV-TRBA, as a control for nonspecific activation of the CD8+T cells). Specifically, ARPE-19 cells were plated in a 96-well plate at 104cells / well and infected with the GFP-expressing human CMV strain TB40 / E at multiplicity of 2 infectious units per cell. After 48 hours, combinations of antibodies and / or purified CD8+T cells (105cells / well) were added. After 24 to 48 hours, of incubation, each well was analyzed by fluorescence microscopy to evaluate for GFP expression as a marker of CMV-infected cells. Quantification was performed by counting green pixels in each image using computer software.

[0089] REFERENCES

[0090] The following references are herein incorporated by reference in their entirety with the exception that, should the scope and meaning of a term conflict with a definition explicitly set forth herein, the definition explicitly set forth herein controls: Elshiaty et al. (2021) Int J Mol Sci 22(11):5632, doi: 10.3390 / ijms22115632Steinmetz et al. (2016) MAbs 8(5):867-78, doi: 10.1080 / 19420862.2016.1162932Li et al. (2005) Immunology 116(4):487-98, doi: 10.1111 / j.1365-2567.2005.02247.xXu et al. (2017) Science 358(6359):85-90. doi: 10.1126 / science.aan8630WO20 17044895 - MVA-gH / gL-PC Vaccine Derived Antibodies Neutralizing Human Cytomegalovirus Infectivity and Methods Thereof

[0091] All scientific and technical terms used in this application have meanings commonly used in the art unless otherwise specified.

[0092] As used herein, the terms “subject”, “patient”, and “individual” are used interchangeably to refer to humans and non-human animals. The terms “non-human animal” and “animal” refer to all non-human vertebrates, e.g., non-human mammals and non-mammals, such as non-human primates, horses, sheep, dogs, cows, pigs, chickens, and other veterinary subjects and test animals. In some embodiments, the subject is a mammal. In some embodiments, the subject is a human.

[0093] As used herein, the term “diagnosing” refers to the physical and active step of informing, i.e., communicating verbally or by writing (on, e.g., paper or electronic media), another party, e.g., a patient, of the diagnosis. Similarly, “providing a prognosis” refers to the physical and active step of informing, i.e., communicating verbally or by writing (on, e.g., paper or electronic media), another party, e.g., a patient, of the prognosis.

[0094] The use of the singular can include the plural unless specifically stated otherwise. As used in the specification and the appended claims, the singular forms “a”, “an”, and “the” can include plural referents unless the context clearly dictates otherwise.

[0095] As used herein, “and / or” means “and” or “or”. For example, “A and / or B” means “A, B, or both A and B” and “A, B, C, and / or D” means “A, B, C, D, or a combination thereof’ and said “A, B, C, D, or a combination thereof’ means any subset of A, B, C, and D, for example, a single member subset (e.g., A or B or C or D), a two-membersubset (e.g., A and B; A and C; etc.), or a three-member subset (e.g., A, B, and C; or A, B, and D; etc.), or all four members (e.g., A, B, C, and D).

[0096] As used herein, the phrase “one or more of’, e.g., “one or more of A, B, and / or C” means “one or more of A”, “one or more of B”, “one or more of C”, “one or more of A and one or more of B”, “one or more of B and one or more of C”, “one or more of A and one or more of C” and “one or more of A, one or more of B, and one or more of C”.

[0097] The phrase “comprises or consists of A” is used as a tool to avoid excess page and translation fees and means that in some embodiments the given thing at issue: comprises A or consists of A. For example, the sentence “In some embodiments, the composition comprises or consists of A” is to be interpreted as if written as the following two separate sentences: “In some embodiments, the composition comprises A. In some embodiments, the composition consists of A.”

[0098] Similarly, a sentence reciting a string of alternates is to be interpreted as if a string of sentences were provided such that each given alternate was provided in a sentence by itself. For example, the sentence “In some embodiments, the composition comprises A, B, or C” is to be interpreted as if written as the following three separate sentences: “In some embodiments, the composition comprises A. In some embodiments, the composition comprises B. In some embodiments, the composition comprises C ” As another example, the sentence “In some embodiments, the composition comprises at least A, B, or C” is to be interpreted as if written as the following three separate sentences: “In some embodiments, the composition comprises at least A. In some embodiments, the composition comprises at least B. In some embodiments, the composition comprises at least C ”

[0099] As used herein, the terms “protein”, “polypeptide” and “peptide” are used interchangeably to refer to two or more amino acids linked together. Groups or strings of amino acid abbreviations are used to represent peptides. Except when specifically indicated, peptides are indicated with the N-terminus on the left and the sequence is written from the N-terminus to the C-terminus. Except when specifically indicated, peptides are indicated with the N-terminus on the left and the sequences are written from the N-terminus to the C-terminus. Similarly, except when specifically indicated, nucleic acid sequences are indicated with the 5’ end on the left and the sequences are written from 5’ to 3’.

[0100] With respect to the CODV-TRBAs herein, the term “light chain” refers to a polypeptide that comprises an Anti-CD3 VL sequence and an Anti-CMV VL sequence, which said polypeptide need not contain an antibody light chain constant region. Withrespect to the CODV-TRBAs herein, the term “heavy chain” refers to a polypeptide that comprises an Anti-CD3 VH sequence and an Anti-CMV VH sequence, which said polypeptide need not contain an antibody heavy chain constant region.

[0101] As used herein, a given percentage of “sequence identity” refers to the percentage of nucleotides or amino acid residues that are the same between sequences, when compared and optimally aligned for maximum correspondence over a given comparison window, as measured by visual inspection or by a sequence comparison algorithm in the art, such as the BLAST algorithm, which is described in Altschul et al., (1990) J Mol Biol 215:403-410. Software for performing BLAST (e.g., BLASTP and BLASTN) analyses is publicly available through the National Center for Biotechnology Information (ncbi.nlm.nih.gov). The comparison window can exist over a given portion, e.g., a functional domain, or an arbitrarily selection a given number of contiguous nucleotides or amino acid residues of one or both sequences. Alternatively, the comparison window can exist over the full length of the sequences being compared. For purposes herein, where a given comparison window (e.g., over 80% of the given sequence) is not provided, the recited sequence identity is over 100% of the given sequence.Additionally, for the percentages of sequence identity of the proteins provided herein, the percentages are determined using BLASTP 2.8.0+, scoring matrix BLOSUM62, and the default parameters available at blast.ncbi.nlm.nih.gov / Blast.cgi. See also Altschul, et al., (1997) Nucleic Acids Res 25:3389-3402; and Altschul, et al., (2005) FEBS J 272:5101- 5109. An “amino acid difference” refers to an amino acid substitution, addition, or deletion.

[0102] Optimal alignment of sequences for comparison can be conducted, e.g., by the local homology algorithm of Smith & Waterman, Adv Appl Math 2:482 (1981), by the homology alignment algorithm of Needleman & Wunsch, J Mol Biol 48:443 (1970), by the search for similarity method of Pearson & Lipman, PNAS USA 85:2444 (1988), by computerized implementations of these algorithms (GAP, BESTFIT, FASTA, and TFASTA in the Wisconsin Genetics Software Package, Genetics Computer Group, 575 Science Dr., Madison, WI), or by visual inspection.

[0103] To the extent necessary to understand or complete the disclosure of the present invention, all publications, patents, and patent applications mentioned herein are expressly incorporated by reference therein to the same extent as though each were individually so incorporated.

[0104] Having thus described exemplary embodiments of the present invention, it should be noted by those skilled in the art that the within disclosures are exemplary only andthat various other alternatives, adaptations, and modifications may be made within the scope of the present invention. Accordingly, the present invention is not limited to the specific embodiments as illustrated herein, but is only limited by the following claims.

Claims

What is claimed is:

1. A cross-over dual variable T cell directing bispecific antibody (CODV-TRBA), which comprises a first chain and a second chain, said first chain comprises a light chain variable domain of an anti-CD3 antibody (Anti- CD3 VL) and a light chain variable domain of an anti-cytomegalovirus (CMV) antibody (Anti- CMV VL); and said second chain comprises a heavy chain variable domain of the anti-CD3 antibody (Anti-CD3 VH) and a heavy chain variable domain of the anti-CMV antibody (Anti-CMV VH), wherein(a) the Anti-CMV VL is linked to C-terminus of the Anti-CD3 VL and the Anti-CD3 VH is linked to the C-terminus of the Anti-CMV VH; or(b) the Anti-CD3 VL is linked to C-terminus of the Anti-CMV VL and the Anti-CMV VH is linked to the C-terminus of the Anti-CD3 VH.

2. The CODV-TRBA according to claim 1, wherein the CDR sequences of the anti-CD3 antibody and the anti-CMV antibody are:- Anti-CD3 CDRLL RASSSVSYMH ( SEQ ID NO: 1 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL2: ATSNLAS ( SEQ ID NO: 2 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRL3: QQWSSNPPT ( SEQ ID NO: 3 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRL1 : QSVSND ( SEQ ID NO: 4 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRL2: YAS ( SEQ ID NO: 5 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRL3: QQDYSSPWT ( SEQ ID NO: 6 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH1: GYT FT I YG ( SEQ ID NO: 7 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH2: INTYTGEP ( SEQ ID NO: 8 ) , with up to 1 or 2 amino acid differences;- Anti-CMV CDRH3: ARKGYYGSSGYFDY ( SEQ ID NO: 9 ) , with up to 1, 2, or 3 amino acid differences;- Anti-CD3 CDRH1: SYTMH ( SEQ ID NO: 10 ) , with up to 1 or 2 amino acid differences;- Anti-CD3 CDRH2: YINPSSGYTKYNQKFKD ( SEQ ID NO: 11 ) , with up to 1, 2, or 3 amino acid differences; and- Anti-CD3 CDRH3: WQDYDVYFDY ( SEQ ID NO: 12 ) , with up to 1 or 2 amino acid differences; wherein up to 8 of the CDR sequences have amino acid differences and / or the total number of amino acid differences in the CDR sequences is not more than 10.

3. The CODV-TRBA according to claims 1 or claim 2, wherein the CDR sequences of the anti- CD3 antibody and the anti-CMV antibody are:- Anti-CD3 CDRL1: RASSSVSYMH (SEQ IDNO: 1);- Anti-CD3 CDRL2: ATSNLAS (SEQ ID NO: 2 ) ;- Anti-CD3 CDRL3: QQWSSNPPT (SEQ IDNO: 3);- Anti-CMV CDRL1 : QSVSND ( SEQ ID NO: 4 ) ;- Anti-CMV CDRL2: YAS ( SEQ ID NO: 5 ) ;- Anti-CMV CDRL3: QQDYSSPWT (SEQ ID NO: 6) ;- Anti-CMV CDRH1: GYT FT I YG (SEQ IDNO: 7);- Anti-CMV CDRH2: INTYTGEP (SEQ ID NO: 8 ) ;- Anti-CMV CDRH3: ARKGYYGSSGYFDY (SEQ ID NO: 9);- Anti-CD3 CDRH1: SYTMH (SEQ ID NO: 10) ;- Anti-CD3 CDRH2: YINPSSGYTKYNQKFKD (SEQ ID NO: 11) ; and- Anti-CD3 CDRH3: WQDYDVYFDY (SEQ ID NO: 12) .

4. The CODV-TRBA according to any one of claims 1 - 3, wherein (a) the sequences of framework regions of the Anti-CD3 VL and the Anti-CMV VL have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% over the combination of SEQ ID NO: 13 to SEQ ID NO: 20, and / or (b) the sequences of framework regions of the Anti-CD3 VH and the Anti-CMV VH have a collective sequence identity of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% over the combination of SEQ ID NO: 21 to SEQ ID NO: 28.

5. The CODV-TRBA according to any one of claims 1 - 4, wherein (a) the Anti-CD3 VL FR1 sequence comprises or consists of SEQ ID NO: 13 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR2 sequence comprises or consists of SEQ ID NO: 14 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR3 sequence comprises or consists of SEQ ID NO: 15 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VL FR4 sequence comprises or consists of SEQ ID NO: 16 with up to 1 or 2 amino acid differences; theAnti-CMV VL FR1 sequence comprises or consists of SEQ ID NO: 17 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR2 sequence comprises or consists of SEQ ID NO: 18 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR3 sequence comprises or consists of SEQ ID NO: 19 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VL FR4 sequence comprises or consists of SEQ ID NO: 20 with up to 1 or 2 amino acid differences; and (b) the Anti-CMV VH FR1 sequence comprises or consists of SEQ ID NO: 21 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR2 sequence comprises or consists of SEQ ID NO: 22 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR3 sequence comprises or consists of SEQ ID NO: 23 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CMV VH FR4 sequence comprises or consists of SEQ ID NO: 24 with up to 1, 2, or 3 amino acid differences; the Anti-CD3 VH FR1 sequence comprises or consists of SEQ ID NO: 25 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VH FR2 sequence comprises or consists of SEQ ID NO: 26 with up to 1, 2, 3, 4, or 5 amino acid differences; the Anti-CD3 VH FR3 sequence comprises or consists of SEQ ID NO: 27 with up to 1, 2, 3, 4, or 5 amino acid differences; and / or the Anti-CD3 VH FR4 sequence comprises or consists of SEQ ID NO: 28 with up to 1, 2, or 3 amino acid differences.

6. The CODV-TRBA according to any one of claims 1 - 5, wherein (a) the Anti-CD3 VL sequence comprises or consists of SEQ ID NO: 42 and / or the Anti-CD3 VH sequence comprises or consists of SEQ ID NO: 44; and (b) the Anti-CMV VL sequence comprises or consists of SEQ ID NO: 43 and / or the Anti-CMV VH sequence comprises or consists of SEQ ID NO: 45.

7. The CODV-TRBA according to any one of claims 1 - 6, wherein the first chain comprises an antibody light chain constant region at its C-terminus and / or the second chain comprises an antibody heavy chain constant region at its C-terminus.

8. The CODV-TRBA according to claim 7, wherein (a) antibody light chain constant region comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 29, and / or (b) the antibody heavy chain constant region comprises or consists of at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO: 31.

9. The CODV-TRBA according to any one of claims 1 - 8, wherein (a) the amino acid sequence of the first chain is SEQ ID NO: 40, and / or (b) the amino acid sequence of the second chain is SEQ ID NO: 41.

10. The CODV-TRBA according to any one of claims 1 - 8, wherein (a) the amino acid sequence of the first chain is SEQ ID NO: 46, and / or (b) the amino acid sequence of the second chain is SEQ ID NO: 47.

11. A medicament comprising one or more CODV-TRBAs according to any one of claims 1 - 10 and a pharmaceutically acceptable carrier.

12. The medicament according to claim 11, and further comprising CD8+T cells and / or one or more antivirals.

13. A method of killing cells infected with a cytomegalovirus (CMV) or treating a subject for an infection by the CMV, which comprises administering to the cells or the subject one or more CODV-TRBAs according to any one of claims 1 - 10 or a medicament according to claim 11 or 12.

14. The method according to claim 13, which further comprises administering CD8+T cells to the cells or the subject.

15. A kit comprising one or more CODV-TRBAs according to any one of claims 1 - 10 packaged together with CD8+T cells and / or one or more antivirals.

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