RNA compositions targeting HIV

Abstract

The present disclosure provides compositions (e.g., pharmaceutical compositions) for delivering anti-HIV antibody agents and related techniques (e.g., components thereof and / or methods related thereto). In addition to this, the present disclosure also provides polyribonucleotides encoding an immunoglobulin chain of an anti-HIV antibody agent.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 63 / 342,057, filed May 13, 2022, the contents of which are hereby incorporated by reference in their entirety. Background Art

[0003] Human immunodeficiency virus (HIV) is a contagious virus associated with acquired immune deficiency syndrome (AIDS). According to the World Health Organization, over 37 million people worldwide are currently infected with HIV. In 2020, approximately 680,000 people died from HIV-related causes and approximately 1.5 million people acquired HIV. Currently, there is no cure for HIV. Summary of the Invention

[0004] The present disclosure recognizes that HIV mutates rapidly. The high variability of HIV viruses enables them to evade host immunity and / or therapeutic pressure. To combat viral escape, combination therapies targeting HIV have been considered, including anti-HIV antibody agents. However, the development of such combination therapies has been hampered by many challenges. First, developing individual HIV therapies is time-consuming and expensive. For example, the development of anti-HIV antibodies is challenged by the high demand and high cost of production, including purification and formulation methods associated with protein therapeutics. Second, combination therapies pose regulatory challenges. In addition to ensuring that combination therapies will be safe and effective, strict regulation of the manufacture of individual therapies, the mixing of multiple therapies, and quality control during storage and administration also complicates the use of combination therapies. Third, administering antibodies to subjects can be painful and time-consuming. Generally, antibodies are administered intravenously over a long period of time. The administration of multiple antibodies can increase the complexity of antibody administration, thereby increasing patient discomfort and consuming additional time. Finally, the serum half-life of recombinant antibodies can be short.

[0005] The present disclosure provides insights into addressing these challenges, making it possible to safely, reliably, and potently deliver not only a single anti-HIV antibody agent to a subject, but also to deliver multiple anti-HIV therapeutic agents, including multiple anti-HIV antibody agents, to a subject. For example, the present disclosure describes an antibody agent or portion thereof (e.g., an immunoglobulin chain) delivered to a subject via a polyribonucleotide. An antibody agent delivered to a subject as one or more polyribonucleotides encoding an antibody agent is referred to herein as a "RiboMab." After one or more polyribonucleotides encoding an antibody agent are delivered to a subject, the subject's body expresses the antibody agent, i.e., a RiboMab." Additionally, the term "RibobNAb" refers to a RiboMab comprising all or part of a broadly neutralizing antibody (bNAb), such as a broadly neutralizing antibody targeting HIV. Utilizing polyribonucleotides as therapeutic agents (as opposed to administering the antibody agent itself) involves a simpler and less expensive manufacturing process. The production of polyribonucleotides encoding antibody agents (e.g., anti-HIV antibody agents) is less complex, and manufacturing can be simplified (e.g., by circumventing the need for intensive glycan production and analysis), thereby alleviating risks associated with Regulatory and production challenges associated with the development and use of antibody agents themselves. In addition, polyribonucleotides effectively produce effects similar to recombinant proteins, but often require administration to subjects in much smaller amounts. This is because polyribonucleotides encoding, for example, anti-HIV antibody agents can be administered to subjects and the subject's body itself produces the anti-HIV antibody agent. Using smaller amounts can provide patients with a more pleasant experience and increase patient compliance with treatment regimens. The present disclosure also provides a technology that addresses certain limitations of recombinant antibody technology by utilizing RNA technology as a way to directly express antibody agents in patients' cells, including, for example, the short serum half-life of recombinant antibodies.

[0006] RiboMab technology also allows for the simultaneous administration of two or more antibody agents to a subject. Typically, antibodies produced by, for example, humans, contain four polypeptide chains—two “heavy” chains and two “light” chains. Each polypeptide chain (whether heavy or light) includes (1) a “variable” domain, whose sequence varies between antibodies and whose structure determines the antigen to which the antibody binds, and (2) a “constant” domain, whose sequence and structure generally remain constant within a given class of antibodies and therefore have little effect on antigen binding. In humans, specialized white blood cells called “B cells” produce antibodies. Heavy and light chains are assembled to form antibodies through two key pairings: (1) the fragment crystallizable (Fc) domains of the two heavy chains pair together, and (2) the two light chains each pair with a heavy chain via a disulfide bond. During normal antibody production in humans, a single B cell produces a single antibody. In this case, the correct pairing of heavy and light chains is ensured because only one heavy chain and one light chain are present in each B cell. However, administration of a nucleic acid composition encoding more than one antibody agent to a subject requires proper assembly of immunoglobulin chains (eg, heavy and light chains) so that unwanted byproducts (eg, antibody agents with unintended pairings) are not formed.

[0007] In addition, the present disclosure also provides polyribonucleotides encoding immunoglobulin chains of antibody agents.

[0008] In one aspect, the immunoglobulin chain comprises a heavy chain variable (VH) domain. In some embodiments, the VH domain comprises a heavy chain complementarity determining region (HCDR) 1 comprising the amino acid sequence of SEQ ID NO: 6; a HCDR2 comprising the amino acid sequence of SEQ ID NO: 9; and a HCDR3 comprising the amino acid sequence of SEQ ID NO: 12.

[0009] In some embodiments, the VH domain comprises or consists of an amino acid sequence according to SEQ ID NO: 24. In some embodiments, the VH domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to an amino acid sequence according to SEQ ID NO: 24.

[0010] In some embodiments, the polyribonucleotide comprises a VH domain encoding sequence. In some embodiments, the VH domain encoding sequence comprises (a) a HCDR1 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 7, (b) a HCDR2 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 10, and (c) a HCDR3 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 13.

[0011] In some embodiments, the VH domain encoding sequence comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 25. In some embodiments, the VH domain encoding sequence comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 25. In some embodiments, the VH domain encoding sequence comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 27. In some embodiments, the VH domain encoding sequence comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 27.

[0012] In some embodiments, an immunoglobulin chain comprising a VH domain as described herein comprises one or more constant domains. In some embodiments, the VH domain is operably linked to one or more constant domains.

[0013] In some embodiments, an immunoglobulin chain comprising a VH domain as described herein comprises one or more constant domains. In some embodiments, the VH domain is operably linked to one or more constant domains.

[0014] In some embodiments, the one or more constant domains comprise a CH2 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain.

[0015] In some embodiments, the one or more constant domains comprise a CH3 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain.

[0016] In some embodiments, one or more constant domains comprise a hinge domain.In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain.

[0017] In some embodiments, the one or more constant domains comprise a CH1 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain.

[0018] In some embodiments, the one or more constant domains comprise a CL domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CL domain.

[0019] In some embodiments, an immunoglobulin chain comprising a VH domain as described herein comprises a CH1 domain, a hinge domain, a CH2 domain, and a CH3 domain.

[0020] In some embodiments, an immunoglobulin chain comprising a VH domain as described herein comprises a CL domain, a hinge domain, a CH2 domain, and a CH3 domain.

[0021] The present disclosure also provides a polyribonucleotide encoding an immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a light chain variable (VL) domain.

[0022] In some embodiments, the VL domain comprises (a) a light chain complementarity determining region (LCDR) 1 comprising the amino acid sequence according to SEQ ID NO: 15; (b) LCDR2 comprising the amino acid sequence according to SEQ ID NO: 18 (GTS); and (c) LCDR3 comprising the amino acid sequence according to SEQ ID NO: 21.

[0023] In some embodiments, the polyribonucleotide comprises a VL domain coding sequence. In some embodiments, the VL domain coding sequence comprises (a) a LCDR1 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 16; a LCDR2 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 19 (GGCACCAGC); and a LCDR3 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 22.

[0024] In some embodiments, the VL domain comprises or consists of an amino acid sequence according to SEQ ID NO: 29. In some embodiments, the VL domain encoding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 30.

[0025] In some embodiments, the immunoglobulin chain comprising the VL domain also comprises a constant domain. In some embodiments, the VL domain is operably connected to the constant domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the constant domain.

[0026] In some embodiments, the immunoglobulin chain comprising the VL domain further comprises a CL domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CL domain. In some embodiments, the CL domain is a kappa constant domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain.

[0027] In some embodiments, the immunoglobulin chain comprising the VL domain further comprises a CH1 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH1 domain.

[0028] In addition, the present disclosure also provides a polyribonucleotide encoding an immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a heavy chain variable (VH) domain and a light chain variable (VL) domain. In some embodiments, the VH domain comprises HCDR1 comprising an amino acid sequence according to SEQ ID NO: 6; HCDR2 comprising an amino acid sequence according to SEQ ID NO: 9; and HCDR3 comprising an amino acid sequence according to SEQ ID NO: 12. In some embodiments, the VL domain comprises LCDR1 comprising an amino acid sequence according to SEQ ID NO: 15; LCDR2 comprising an amino acid sequence according to SEQ ID NO: 18 (GTS); and LCDR3 comprising an amino acid sequence according to SEQ ID NO: 21.

[0029] In some embodiments, the polyribonucleotide comprises a VH domain encoding sequence and a VL domain encoding sequence. In some embodiments, the VH domain encoding sequence comprises a HCDR1 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 7; a HCDR2 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 10; and a HCDR3 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 13. In some embodiments, the VL domain encoding sequence comprises a LCDR1 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 16; a LCDR2 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 19 (GGCACCAGC); and a LCDR3 encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 22.

[0030] In some embodiments, the immunoglobulin chain comprises a single-chain variable fragment (scFv).In some embodiments, the scFv comprises a VH domain, a linker, and a VL domain.

[0031] In some embodiments, the scFv comprises, in order, a VH domain, a linker, and a VL domain. In some embodiments, the scFv comprises, in order, a VH domain comprising or consisting of an amino acid sequence according to SEQ ID NO: 24; a linker; and a VL domain comprising or consisting of an amino acid sequence according to SEQ ID NO: 29.

[0032] In some embodiments, the scFv comprises, in order, a VL domain, a linker, and a VH domain. In some embodiments, the scFv comprises, in order, a VL domain comprising or consisting of an amino acid sequence according to SEQ ID NO: 29; a linker; and a VH domain comprising or consisting of an amino acid sequence according to SEQ ID NO: 24.

[0033] In some embodiments, the linker comprises an amino acid sequence according to SEQ ID NO: 32. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a linker and comprising or consisting of a sequence according to SEQ ID NO: 33.

[0034] In some embodiments, the linker comprises an amino acid sequence according to SEQ ID NO: 35. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a linker and comprising or consisting of a sequence according to SEQ ID NO: 36.

[0035] In some embodiments, the immunoglobulin chain comprising a VH domain and a VL domain as described herein comprises one or more constant domains. In some embodiments, the VH domain and the VL domain are operably linked to one or more constant domains.

[0036] In some embodiments, the immunoglobulin chain comprises one or more constant domains, and the hinge domain is between the scFv and the one or more constant domains.

[0037] In some embodiments, the one or more constant domains comprise a CH2 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain.

[0038] In some embodiments, the one or more constant domains comprise a CH3 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain.

[0039] In some embodiments, one or more constant domains comprise a hinge domain.In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain.

[0040] In some embodiments, the one or more constant domains comprise a CH1 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain.

[0041] According to the present disclosure, the CH2 domain of any of the above embodiments may comprise an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 53. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 53.

[0042] In some embodiments, the ribonucleic acid sequence encoding the CH2 domain comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 54. In some embodiments, the ribonucleic acid sequence encoding the CH2 domain comprises or consists of the sequence according to SEQ ID NO: 54.

[0043] In some embodiments, the CH2 domain comprises one or more substitution mutations. In some embodiments, the one or more substitution mutations in the CH2 domain comprise G236A, A330L, I332E, or a combination thereof or consist thereof, and wherein the substitution mutation position is according to EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise G236A or consist thereof, and wherein the substitution mutation position is according to EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise I332E or consist thereof, and wherein the substitution mutation position is according to EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise G236A and I332E or consist thereof, and wherein the substitution mutation position is according to EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise G236A, A330L, and I332E or consist thereof, and wherein the substitution mutation position is according to EU numbering.

[0044] In some embodiments, the CH2 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 56. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 56.

[0045] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain, which has a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 57. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain and comprising or consisting of a sequence according to SEQ ID NO: 57.

[0046] In some embodiments, the CH2 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 59. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 59.

[0047] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 60. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain and comprising or consisting of a sequence according to SEQ ID NO: 60.

[0048] In some embodiments, the CH2 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 62. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 62.

[0049] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 63. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain and comprising or consisting of a sequence according to SEQ ID NO: 63.

[0050] In some embodiments, the CH2 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 65. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 65.

[0051] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the ribonucleic acid sequence according to SEQ ID NO: 66. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain and comprising or consisting of a sequence according to SEQ ID NO: 66.

[0052] According to the present disclosure, the CH3 domain of any of the above embodiments may comprise an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 68. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 68.

[0053] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes a CH3 domain and is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 69. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes a CH3 domain and comprises or consists of the sequence according to SEQ ID NO: 69.

[0054] In some embodiments, the CH3 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 71. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 71.

[0055] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes a CH3 domain and is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 72. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes a CH3 domain and comprises or consists of the sequence according to SEQ ID NO: 72.

[0056] In some embodiments, the CH3 domain comprises one or more substitution mutations. In some embodiments, the one or more substitution mutations in the CH3 domain comprise or consist of M428L, N434S, or a combination thereof, and wherein the substitution mutation positions are according to EU numbering.

[0057] In some embodiments, the CH3 domain comprises or consists of an amino acid sequence according to SEQ ID NO: 74. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH3 domain and comprising or consisting of a sequence according to SEQ ID NO: 75.

[0058] In some embodiments, the CH3 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 77. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 77.

[0059] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the ribonucleic acid sequence according to SEQ ID NO: 78. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and comprising or consisting of the sequence according to SEQ ID NO: 78.

[0060] In some embodiments, the one or more substitution mutations in the CH3 domain comprise or consist of Y349C, T366S, L368A, Y407V, or a combination thereof, and wherein the substitution mutation positions are according to EU numbering. In some embodiments, the one or more substitution mutations in the CH3 domain comprise or consist of Y349C, T366S, L368A, Y407V, M428L, N434S, or a combination thereof, and wherein the substitution mutation positions are according to EU numbering. In some embodiments, the one or more substitution mutations in the CH3 domain comprise or consist of Y349C, T366S, L368A, Y407V, M428L, and N434S, and wherein the substitution mutation positions are according to EU numbering.

[0061] In some embodiments, the CH3 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 80. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 80.

[0062] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the ribonucleic acid sequence according to SEQ ID NO: 81. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and comprising or consisting of a sequence according to SEQ ID NO: 81.

[0063] In some embodiments, the CH3 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 83. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 83.

[0064] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the ribonucleic acid sequence according to SEQ ID NO: 84. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and comprising or consisting of a sequence according to SEQ ID NO: 84.

[0065] In some embodiments, the one or more substitution mutations in the CH3 domain comprise S354C, T366W, or a combination thereof, or consist thereof, and wherein the substitution mutation positions are according to EU numbering. In some embodiments, the one or more substitution mutations in the CH3 domain comprise S354C, T366W, M428L, N434S, or a combination thereof, or consist thereof, and wherein the substitution mutation positions are according to EU numbering. In some embodiments, the one or more substitution mutations in the CH3 domain comprise S354C, T366W, M428L, and N434S, or consist thereof, and wherein the substitution mutation positions are according to EU numbering.

[0066] In some embodiments, the CH3 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 86. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 86.

[0067] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes a CH3 domain and is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to a ribonucleic acid sequence according to SEQ ID NO: 87. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes a CH3 domain and comprises or consists of a sequence according to SEQ ID NO: 87.

[0068] In some embodiments, the CH3 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 89. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 89.

[0069] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 90. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain and comprising or consisting of a sequence according to SEQ ID NO: 90.

[0070] According to the present disclosure, the hinge domain of any of the above embodiments may comprise an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 104. In some embodiments, the hinge domain comprises or consists of the amino acid sequence according to SEQ ID NO: 104.

[0071] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 105. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain and comprising or consisting of a sequence according to SEQ ID NO: 105.

[0072] In some embodiments, the hinge domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 110. In some embodiments, the hinge domain comprises or consists of the amino acid sequence according to SEQ ID NO: 110.

[0073] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 111. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain and comprising or consisting of a sequence according to SEQ ID NO: 111.

[0074] In some embodiments, the hinge domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 107. In some embodiments, the hinge domain comprises or consists of the amino acid sequence according to SEQ ID NO: 107.

[0075] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the ribonucleic acid sequence according to SEQ ID NO: 108. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain and comprising or consisting of a sequence according to SEQ ID NO: 108.

[0076] According to the present disclosure, the CH1 domain of any of the above embodiments may comprise an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 38. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 38.

[0077] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 39. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 39.

[0078] In some embodiments, the CH1 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 41. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 41.

[0079] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 42. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 42.

[0080] In some embodiments, the CH1 domain comprises one or more substitution mutations. In some embodiments, the one or more substitution mutations in the CH1 domain comprise or consist of K147E, K213D, or a combination thereof, and wherein the substitution mutation positions are according to EU numbering.

[0081] In some embodiments, the CH1 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 50. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 50.

[0082] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 51. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 51.

[0083] In some embodiments, the CH1 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 44. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 44.

[0084] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 45. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 45.

[0085] In some embodiments, the CH1 domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 47. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 47.

[0086] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 48. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 48.

[0087] According to the present disclosure, the CL domain of any of the above embodiments may comprise an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 92. In some embodiments, the CL domain of any of the above embodiments may comprise or consist of the amino acid sequence according to SEQ ID NO: 92.

[0088] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CL domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the ribonucleic acid sequence according to SEQ ID NO: 93. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CL domain and comprising or consisting of a sequence according to SEQ ID NO: 93.

[0089] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain CL domain comprising one or more substitution mutations. In some embodiments, the one or more substitution mutations in the CL domain comprise Q124E or consist of it, and wherein the substitution mutation position is according to EU numbering. In some embodiments, the one or more substitution mutations in the CL domain comprise R108A, T109S or a combination thereof or consist of it, and wherein the substitution mutation position is according to EU numbering. In some embodiments, the one or more substitution mutations in the CL domain comprise R108A, T109S, Q124E or a combination thereof or consist of it, and wherein the substitution mutation position is according to EU numbering.

[0090] In some embodiments, the CL domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 95. In some embodiments, the CL domain comprises or consists of the amino acid sequence according to SEQ ID NO: 95.

[0091] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 96. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain and comprising or consisting of a sequence according to SEQ ID NO: 96.

[0092] In some embodiments, the CL domain comprises one or more substitution mutations. In some embodiments, the one or more substitution mutations in the CL domain comprise or consist of E123K, Q124R, or a combination thereof, and wherein the substitution mutation positions are according to EU numbering.

[0093] In some embodiments, the CL domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 98. In some embodiments, the CL domain comprises or consists of the amino acid sequence according to SEQ ID NO: 98.

[0094] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to a ribonucleic acid sequence according to SEQ ID NO: 99. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain and comprising or consisting of a sequence according to SEQ ID NO: 99.

[0095] In some embodiments, the one or more substitution mutations in the CL domain comprise or consist of E123R, Q124K, or a combination thereof, and wherein the substitution mutation positions are according to EU numbering.

[0096] In some embodiments, the CL domain comprises an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 101. In some embodiments, the CL domain comprises or consists of the amino acid sequence according to SEQ ID NO: 101.

[0097] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain and having at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identity to the ribonucleic acid sequence according to SEQ ID NO: 102. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain and comprising or consisting of a sequence according to SEQ ID NO: 102.

[0098] In some embodiments, the polyribonucleotides as provided herein encode immunoglobulin chains, wherein:

[0099] (i) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain, and the CH2 domain comprises one or more substitution mutations, wherein the one or more substitution mutations comprise or consist of G236A, A330L, I332E, or a combination thereof,

[0100] (ii) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain, and the CH3 domain comprises one or more substitution mutations, wherein the one or more substitution mutations comprise or consist of Y349C, S354C, T366S, T366W, L368A, Y407V, M428L, N434S, or a combination thereof,

[0101] (iii) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain, and the CH1 domain comprises one or more substitution mutations, wherein the one or more substitution mutations comprise or consist of K147E, K213D, or a combination thereof,

[0102] (iv) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CL domain, and the CL domain comprises one or more substitution mutations, wherein the one or more substitution mutations comprise or consist of R108A, T109S, E123K, E123R, Q124E, Q124K, Q124R, or a combination thereof, or

[0103] (v) combinations thereof;

[0104] The substitution mutation positions are numbered according to EU.

[0105] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 614. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 614.

[0106] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence of SEQ ID NO: 613. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence of SEQ ID NO: 613.

[0107] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 617. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 617.

[0108] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 616. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 616.

[0109] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 623. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 623.

[0110] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 622. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 622.

[0111] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 626. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 626.

[0112] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 625. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 625.

[0113] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 635. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 635.

[0114] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 634. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 634.

[0115] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 641. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 641.

[0116] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 640. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 640.

[0117] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 644. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 644.

[0118] In some embodiments, the polyribonucleotides as provided herein comprise a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence of SEQ ID NO: 643. In some embodiments, the polyribonucleotides as provided herein comprise or consist of the ribonucleic acid sequence of SEQ ID NO: 643.

[0119] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 647. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 647.

[0120] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 646. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 646.

[0121] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 650. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 650.

[0122] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 649. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 649.

[0123] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 653. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 653.

[0124] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 652. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 652.

[0125] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 629. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 629.

[0126] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 628. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 628.

[0127] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 620. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 620.

[0128] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 619. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 619.

[0129] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 638. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 638.

[0130] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 637. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 637.

[0131] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 668. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 668.

[0132] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 667. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 667.

[0133] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 632. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 632.

[0134] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 631. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 631.

[0135] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 656. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the sequence according to SEQ ID NO: 656.

[0136] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 655. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 655.

[0137] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 659. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the sequence according to SEQ ID NO: 659.

[0138] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 658. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 658.

[0139] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 662. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the sequence according to SEQ ID NO: 662.

[0140] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 661. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 661.

[0141] In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the amino acid sequence according to SEQ ID NO: 665. In some embodiments, the polyribonucleotides as provided herein encode an immunoglobulin chain comprising or consisting of the sequence according to SEQ ID NO: 665.

[0142] In some embodiments, the polyribonucleotide as provided herein comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 664. In some embodiments, the polyribonucleotide as provided herein comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 664.

[0143] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a secretion signal.

[0144] In some embodiments, the secretion signal comprises a ribonucleic acid sequence according to SEQ ID NO:2 or SEQ ID NO:4.

[0145] In some embodiments, the polyribonucleotide comprises one or more non-coding sequence elements.

[0146] In some embodiments, one or more non-coding sequence elements enhance RNA stability and / or translation efficiency.

[0147] In some embodiments, the one or more non-coding sequence elements comprise a 3' untranslated region (UTR), a 5'UTR, a 5'-cap, a polyadenine (polyA) tail, or a combination thereof.

[0148] In some embodiments, the polyA tail is or comprises a modified polyA sequence, preferably an interrupted polyA tail.

[0149] In some embodiments, the polyA tail comprises or consists of a sequence that is at least 90%, at least 95%, or at least 99% identical to SEQ ID NO: 474.

[0150] In some embodiments, the 3'UTR comprises or consists of a nucleic acid sequence that is at least 90%, at least 95%, or at least 99% identical to SEQ ID NO:473.

[0151] In some embodiments, the 5'UTR comprises or consists of a nucleic acid sequence that is at least 90%, at least 95%, or at least 99% identical to SEQ ID NO:472.

[0152] In some embodiments, the 5'-cap is (m27,3'-O)Gppp(m2'-O)ApG.

[0153] In some embodiments, the polyribonucleotide comprises one or more modified ribonucleotides.In some embodiments, the one or more modified ribonucleotides comprise pseudouridine.

[0154] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 454. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 454.

[0155] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 455. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 455.

[0156] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 457. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 457.

[0157] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 456. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 456.

[0158] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 461. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 461.

[0159] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 463. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 463.

[0160] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 465. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 465.

[0161] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 462. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 462.

[0162] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 464. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 464.

[0163] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 466. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 466.

[0164] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 459. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 459.

[0165] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 458. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 458.

[0166] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 467. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 467.

[0167] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 460. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 460.

[0168] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 468. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 468.

[0169] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 470. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 470.

[0170] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 469. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 469.

[0171] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% identical to the ribonucleic acid sequence according to SEQ ID NO: 471. In some embodiments, the polyribonucleotide comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 471.

[0172] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NOs: 669-864. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to any one of SEQ ID NOs: 669-864.

[0173] In some embodiments, the polyribonucleotide is a non-natural polyribonucleotide.

[0174] In some embodiments, the polyribonucleotide is an engineered polyribonucleotide.

[0175] In some embodiments, the polyribonucleotide is an isolated polyribonucleotide.

[0176] In addition, the present disclosure also provides compositions comprising one or more polyribonucleotides as described herein.

[0177] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 614; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 620.

[0178] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 613; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 619.

[0179] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 617; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 620.

[0180] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 616; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 619.

[0181] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 623; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 620.

[0182] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 622; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 619.

[0183] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 626; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 620.

[0184] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 625; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 619.

[0185] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 635; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 638.

[0186] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 634; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 637.

[0187] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 641; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 638.

[0188] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 640; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 637.

[0189] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 644; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 638.

[0190] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 643; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 637.

[0191] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 647; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 638.

[0192] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 646; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 637.

[0193] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 650; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 638.

[0194] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 649; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 637.

[0195] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 653; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 638.

[0196] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 652; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 637.

[0197] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 635; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 668.

[0198] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 634; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 667.

[0199] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 641; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 668.

[0200] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 640; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 667.

[0201] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 644; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 668.

[0202] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 643; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 667.

[0203] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 647; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 668.

[0204] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 646; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 667.

[0205] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 650; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 668.

[0206] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 649; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 667.

[0207] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 653; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 668.

[0208] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 652; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 667.

[0209] In some embodiments, the composition comprises or consists of: a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 629; and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to the amino acid sequence of SEQ ID NO: 632.

[0210] In some embodiments, the composition comprises or consists of a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 628; and a polyribonucleotide comprising a ribonucleic acid sequence that is at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98% or at least 99% identical to a ribonucleic acid sequence according to any one of SEQ ID NO: 631.

[0211] In some embodiments, the composition further comprises a lipid nanoparticle, a polyplex (PLX), a lipidated polyplex (LPLX) or a liposome, wherein the one or more polyribonucleotides are fully or partially encapsulated within the lipid nanoparticle, polyplex (PLX), lipidated polyplex (LPLX) or the liposome.

[0212] In some embodiments, the composition further comprises a lipid nanoparticle, wherein the one or more polyribonucleotides are encapsulated within the lipid nanoparticle.

[0213] In some embodiments, the lipid nanoparticles are targeted to hepatocytes.

[0214] In some embodiments, lipid nanoparticles are targeted to secondary lymphoid organ cells.

[0215] In some embodiments, lipid nanoparticles are targeted to lung cells.

[0216] In some embodiments, the lipid nanoparticles are cationic lipid nanoparticles.

[0217] In some embodiments, the lipid nanoparticles each comprise a polymer-conjugated lipid, a cationic lipid, and one or more neutral lipids.

[0218] In some embodiments, the polymer-conjugated lipid comprises a PEG-conjugated lipid.

[0219] In some embodiments, the polymer-conjugated lipid comprises 2-[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide.

[0220] In some embodiments, the one or more neutral lipids comprise 1,2-distearoyl-sn-glycero-3-phosphocholine (DPSC).

[0221] In some embodiments, the one or more neutral lipids comprise cholesterol.

[0222] In some embodiments, the cationic lipid comprises ((3-hydroxypropyl)azanediyl)bis(nonane-9,1-diyl)bis(2-butyloctanoate).

[0223] In some embodiments, the lipid nanoparticles each comprise 2-[(polyethylene glycol)-2000]-N,N-ditetradecanoylacetamide, DPSC, cholesterol, and bis(2-butyloctanoate)((3-hydroxypropyl)azanediyl)bis(nonane-9,1-diyl) ester.

[0224] In some embodiments, the lipid nanoparticles comprise about 1-2.5 mol% of the polymer-conjugated lipid of the total lipids, 35-65 mol% of the cationic lipid of the total lipids, and one or more neutral lipids are present at 35-65 mol% of the total lipids.

[0225] In some embodiments, the lipid nanoparticles have an average diameter of about 50-150 nm.

[0226] The present disclosure also provides pharmaceutical compositions. In some embodiments, the pharmaceutical composition comprises a composition provided herein and at least one pharmaceutically acceptable excipient.

[0227] In some embodiments, the medicament comprises a cryoprotectant. In some embodiments, the medicament comprises a buffered aqueous solution.

[0228] In addition, the present disclosure provides methods.

[0229] In some embodiments, the methods comprise administering to a subject a pharmaceutical composition provided herein.

[0230] In some embodiments, the pharmaceutical compositions provided herein are used to treat HIV, comprising administering the pharmaceutical compositions to a subject.

[0231] In some embodiments, the pharmaceutical compositions provided herein are used to prevent HIV, comprising administering the pharmaceutical compositions to a subject.

[0232] In some embodiments, the methods or pharmaceutical compositions provided herein for use comprise administering the pharmaceutical composition to a subject, which results in expression of the immunoglobulin chains of the antibody agent, the antibody agent, or both in the subject.

[0233] In some embodiments, the immunoglobulin chains of the antibody agent, the antibody agent, or both are expressed in the plasma or serum of the subject at a titer of at least 1 μg / ml.

[0234] In some embodiments, the antibody agent exhibits a geometric mean IC50 of less than 0.3 μg / ml for five neutralizing strains of a global reference panel when tested in a TZM-bl cell pseudovirus neutralization assay at antibody agent concentrations up to 25 μg / ml.

[0235] In some embodiments, the antibody agent is capable of neutralizing one or more HIV strains when tested in a TZM-bl cell pseudovirus neutralization assay at antibody agent concentrations up to 25 μg / ml.

[0236] In some embodiments, the antibody agent is capable of neutralizing one or more strains of HIV at a level that is within 3-fold of the level of an equivalent amount of a recombinant benchmark antibody.

[0237] In some embodiments, the recombinant benchmark antibody is an unmodified wild-type IgG antibody comprising the same HCDR1, HCDR2, HCDR2, LCDR1, LCDR2, and LCDR3 as the antibody agent.

[0238] In some embodiments, administering the pharmaceutical composition to the subject comprises administering one or more doses of the pharmaceutical composition to the subject. In some embodiments, one or more doses of the pharmaceutical composition are administered to the subject weekly. In some embodiments, one or more doses of the pharmaceutical composition are administered to the subject every two weeks.

[0239] In some embodiments, the pharmaceutical composition is administered intravenously. In some embodiments, the pharmaceutical composition is administered intramuscularly. In some embodiments, the pharmaceutical composition is administered subcutaneously.

[0240] In some embodiments, the subject has HIV infection or is at risk for developing HSV infection.

[0241] In some embodiments, the method is a method of treating HIV infection.

[0242] In some embodiments, the method is a method of preventing HIV infection.

[0243] Also provided herein are uses of the polyribonucleotides, compositions, and pharmaceutical compositions described herein.

[0244] In some embodiments, provided is a use of a composition or pharmaceutical composition as provided herein for treating HIV in a subject.

[0245] In some embodiments, provided is a use of a composition or pharmaceutical composition as provided herein for preventing HIV in a subject.

[0246] In some embodiments, the subject has HIV infection or is at risk for HSV infection

[0247] In addition, the present disclosure also provides methods for producing antibody agents. In some embodiments, the method comprises administering a composition or pharmaceutical composition as provided herein to a cell, such that the cell expresses and secretes the antibody agent.

[0248] In some embodiments, the cell is a hepatocyte.

[0249] In some embodiments, the cell is in a subject.

[0250] In some embodiments, the cells are ex vivo cells.

[0251] In some embodiments, the antibody agent is produced at a therapeutically relevant plasma concentration or a therapeutically relevant serum concentration. In some embodiments, the therapeutically relevant plasma concentration or the therapeutically relevant serum concentration is at least 1 μg / ml.

[0252] The present disclosure also provides a method for determining one or more characteristics of an antibody agent expressed by a polyribonucleotide, composition or pharmaceutical composition provided herein. In some embodiments, the polyribonucleotide, composition or pharmaceutical composition provided herein is introduced into a cell. In some embodiments, the one or more characteristics comprise: (i) protein expression level of the antibody agent; (ii) binding specificity of the antibody agent to the CD4 binding site of HIV; (iii) efficacy of the antibody agent in mediating target cell death through antibody-dependent cellular cytotoxicity (ADCC); and (iv) efficacy of the antibody agent in mediating target cell death through complement-dependent cytotoxicity (CDC).

[0253] The present disclosure also provides methods comprising contacting a cell with a polyribonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, the method further comprises detecting the antibody agent produced by the cell.

[0254] In some embodiments, the cell is a hepatocyte.

[0255] In some embodiments, the determining step comprises comparing one or more characteristics of the antibody agent to one or more characteristics of a reference antibody that specifically binds to the CD4 binding site of HIV.

[0256] In some embodiments, the determining step comprises assessing the protein expression level of the antibody agent to be above a threshold level.

[0257] In some embodiments, the threshold level is a level sufficient to induce ADCC.

[0258] In some embodiments, the determining step comprises evaluating binding of the antibody agent to the CD4 binding site of HIV.

[0259] In some embodiments, the determining step comprises evaluating the antibody agent in a TZM-bl cell pseudovirus neutralization assay at antibody agent concentrations up to 25 μg / ml.

[0260] In some embodiments, the cell is present in a subject.

[0261] In some embodiments, the cells are ex vivo cells.

[0262] In some embodiments, the one or more characteristics include antibody levels in one or more tissues of the subject.

[0263] The disclosure also provides a manufacturing method. In some embodiments, the method comprises (a) determining one or more features of the polyribonucleotide, composition or pharmaceutical composition provided herein, wherein the one or more features comprise or are composed of:

[0264] (i) length and / or sequence of the polyribonucleotides;

[0265] (ii) polyribonucleotide integrity;

[0266] (iii) the presence and / or location of one or more chemical moieties of the polyribonucleotide;

[0267] (iv) the extent of expression of the antibody agent when the polyribonucleotide is introduced into the cell;

[0268] (v) stability of the polyribonucleotide or its combination;

[0269] (vi) the level of the antibody agent in a biological sample from an organism into which the polyribonucleotide has been introduced;

[0270] (vii) the binding specificity of the antibody agent expressed from the polyribonucleotide, optionally binding to the CD4 binding site of HIV;

[0271] (viii) the efficacy of the antibody agent in mediating target cell death via ADCC;

[0272] (ix) the efficacy of the antibody agent in mediating target cell death through complement-dependent cytotoxicity (CDC);

[0273] (x) the identity and amount / concentration of lipids within the composition;

[0274] (xi) the size of the lipid nanoparticles within the composition;

[0275] (xii) the polydispersity of the lipid nanoparticles within the composition;

[0276] (xiii) the amount / concentration of polyribonucleotides in the composition;

[0277] (xiv) the degree of encapsulation of polyribonucleotides within lipid nanoparticles;

[0278] (xv) levels of double-stranded RNA; and

[0279] (xvi) combinations thereof;

[0280] (B) comparing one or more characteristics of the polyribonucleotide to one or more characteristics of an appropriate reference standard; and

[0281] (C)(i) if the comparison indicates that the polyribonucleotide or composition thereof meets or exceeds the reference standard, designating the polyribonucleotide or composition thereof for one or more additional steps in manufacturing and / or distribution; or

[0282] (C)(ii) taking an alternative action if the comparison indicates that the polyribonucleotide or composition thereof does not meet or exceed the reference standard.

[0283] In some embodiments, polyribonucleotides are assessed and one or more further steps of step (C)(i) are at least or comprise formulation of polyribonucleotides.

[0284] In some embodiments, the composition or pharmaceutical composition is evaluated and one or more further steps of step (C)(i) is or comprises release and distribution of the composition or pharmaceutical composition.

[0285] Thus, the present disclosure provides technologies that allow for the expression of multiple anti-HIV antibodies in a subject, thereby increasing the breadth and potency of the anti-HIV antibodies simultaneously present in the subject and reducing the likelihood of viral escape.

[0286] The provided technology, including exemplary polyribonucleotides, compositions comprising such polyribonucleotides, and methods of making and using such polyribonucleotides, is described in more detail herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0287] Figure 1 shows a schematic diagram of the HIV genome (1A) and the structure of the HIV virion particle (1B). The figure is modified from Musumeci et al., 2015 Molecules, which is incorporated herein by reference in its entirety.

[0288] Figure 2 Potential epitope regions on HIV virion particles to which antibody agents (e.g., broadly neutralizing antibodies (bNAbs) or variants thereof) can bind are shown. The figures are modified from McCoy and Burton, 2017 Immunol Rev., which is incorporated herein by reference in its entirety.

[0289] Figure 3 An exemplary therapeutic strategy utilizing RiboMab technology as described herein to deliver and express anti-HIV RibobNAbs is shown.

[0290] Figure 4 shows exemplary formats of 1-18 RibobNAbs as described herein. Exemplary formats may include IgG (4A), CrossMab CH1-CLx (4B), CrossMab CH1-CLcv (4C) or various orientations / linkers of scFv-Fc RibobNAbs (4D and 4E).

[0291] Figure 5 shows exemplary Fc modifications of 1-18 RibobNAbs as described herein. Exemplary RibobNAb formats can include an unmodified Fc domain (5A), or modifications shown in 5B-D, including GAALIE, / GAIE / GA / IE (5B), L / S (5C), and / or knob-in-hole Ribo bNAbs (5D).

[0292] FIG6 shows a schematic diagram of exemplary polyribonucleotides encoding the heavy chain (6A) and light chain (6B) of an exemplary 1-18 IgG1 antibody agent.

[0293] Figure 7 Shown are the breadth and % potency of various broadly neutralizing HIV antibodies (including 1-18) as tested against 109 pseudoviruses.

[0294] Figure 8 shows schematic diagrams of exemplary polyribonucleotides encoding scFv-Fc 1-18 antibody agents: 1-18scFv-FcVH-LL4 / LL5-VL (8A) and 1-18scFv-Fc VL-LL4 / LL5-VH (8B).

[0295] Figure 9 shows an example 1-18 CrossMab encoding CH1-CLx Schematic representation of exemplary polyribonucleotides of the heavy chain (9A) and light chain (9B) of an antibody agent.

[0296] Figure 10 shows an exemplary 1-18 CrossMab encoding CH1-CLcv Schematic representation of exemplary polyribonucleotides of the heavy chain (10A) and light chain (10B) of an antibody agent.

[0297] Figure 11 Shown are exemplary concentrations (ng / mL) of 1-18 and 1-18 L / S RibobNAbs compared to a control RiboMab as determined by Gyros ELISA in Example 5.

[0298] Figure 12 Shown are exemplary Western blot analyses of 1-18 and 1-18L / S RibobNAbs compared to a control RiboMab under non-reducing and reducing conditions in Example 5.

[0299] Figure 13 Shown are exemplary concentrations of scFv-Fc 1-18L / S RibobNAbs (e.g., VH-LL4-VL, VL-LL5-VH, VH-LL5-VL, and VL-LL4-VH) compared to a control RiboMab and parental IgG as determined by Gyros ELISA in Example 6.

[0300] Figure 14 Shown are exemplary Western blot analyses of scFv-Fc1-18L / S RibobNAbs (e.g., VH-LL4-VL, VL-LL5-VH, VH-LL5-VL, and VL-LL4-VH) compared to control RiboMab and parental IgG under non-reducing conditions in Example 6.

[0301] Figure 15 Shown are exemplary Western blot analyses of scFv-Fc1-18L / S RibobNAbs (e.g., VH-LL4-VL, VL-LL5-VH, VH-LL5-VL, and VL-LL4-VH) compared to control RiboMab and parental IgG under non-reducing conditions in Example 6.

[0302] Figure 16 Shows that CrossMab 1-18L / S RibobNAbs (e.g., CrossMab CH1-CLcv and CrossMab CH1-CLx ) in exemplary concentrations (ng / mL).

[0303] Figure 17As shown in Example 7, CrossMab 1-18L / SRibobNAb (e.g., CrossMab CH1-CLcv and CrossMab CH1-CLx ) were analyzed by Western blot analysis.

[0304] Figure 18 Shown are exemplary in vivo concentrations (ng / mL, logarithmic scale) of 1-18 and 1-18L / S RibobNAbs compared to a control RiboMab as determined by Gyros ELISA in Example 8.

[0305] Figure 19 Exemplary PK of 1-18 IgG, IgG L / S, and scFv-Fc L / S RibobNAbs are shown. 1-18 RibobNAbs were quantified using serum obtained from hFcRnNSG Tg32 mice. The y-axis shows in vivo concentration (μg / mL) on a logarithmic scale. The x-axis shows the time of each blood sampling (hours). Depicted are the results of 1-18VL-LL5-VH L / S scFv-Fc compared to 1-18 and 1-18L / S IgG RibobNAbs as determined by the Gyros ELISA in Example 12. Two different doses of 1-18VL-LL5-VH L / S scFv-Fc (30 μg and 19.56 μg) and 1-18L / S IgG (30 μg and 10 μg) were analyzed.

[0306] Figure 20 Shown are exemplary results of a pseudovirus neutralization test (pVNT) in which TSM.b1 cells were exposed to 1-18 IgG and 1-18 L / S IgG RibobNAbs and pseudoviruses CNE19 (clade B), q23.17 (clade A1), Tro.11 (clade B), RHPA4259.7 (clade B), 6540V4_C1 (clade CRF01_AC), ZM269M.PL1 (clade C), and murine leukemia virus (MuLV) pseudoviruses as negative controls. Results are expressed as IC50 and IC80 values ​​or antibody / IgG concentration, relative luminescence units (RLU) that were reduced by 50% and 80% compared to untreated virus control wells.

[0307] Figure 21Shown are exemplary results of a pseudovirus neutralization test (pVNT) in which TSM.b1 cells were exposed to 1-18 scFv-Fc L / S RibobNAb and pseudoviruses CAP304_2_00_F6_6 (clade C), CE1176_A3 (clade C), 6980.V0.C31 (clade C), 1012_11_TC21_3257 (clade B), PVO.4 (clade B), and murine leukemia virus (MuLV) pseudovirus as negative controls. Results are expressed as IC50 and IC80 values ​​or antibody / IgG concentrations, relative luminescence units (RLU) that were reduced by 50% and 80% compared to untreated virus control wells.

[0308] Figure 22 Shown are exemplary results of a pseudovirus neutralization test (pVNT) in which TSM.b1 cells were exposed to 1-18 scFv-Fc L / S RibobNAb and pseudovirus B), 6540V4_C1 (clade CRF01_AC), ZM269M.PL1 (clade C), and murine leukemia virus (MuLV) pseudovirus as negative controls. Results are expressed as IC50 and IC80 values ​​or antibody / IgG concentrations, relative luminescence units (RLU) that were reduced by 50% and 80% compared to untreated virus control wells.

[0309] Figure 23 Shown are exemplary results of a pseudovirus neutralization test (pVNT) in which TSM.b1 cells were exposed to 1-18 CrossMab RibobNAb and pseudoviruses CNE19 (clade B), q23.17 (clade A1), Tro.11 (clade B), RHPA4259.7 (clade B), R2184_C4 (clade CRF01_AE) and 6540_V4_C1 (clade CRF01_AC) and murine leukemia virus (MuLV) pseudovirus as negative controls. Results are expressed as IC50 and IC80 values ​​or antibody / IgG concentrations, relative luminescence units (RLU) that were reduced by 50% and 80% compared to untreated virus control wells.

[0310] definition

[0311] Compounds of the present disclosure include those generally described above and are further illustrated by the classes, subclasses, and species disclosed herein. As used herein, unless otherwise indicated, the following definitions shall apply. For the purposes of this disclosure, chemical elements are identified according to the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75th edition. In addition, the general principles of organic chemistry are described in "Organic Chemistry", Thomas Sorrell, University Science Books, Sausalito: 1999 and "March's Advanced Organic Chemistry", 5th edition, editors: Smith, MB and March, J., John Wiley & Sons, New York: 2001, the entire contents of which are hereby incorporated by reference.

[0312] Unless otherwise stated, the structure depicted herein is intended to include all stereoisomers (e.g., enantiomers or diastereomers) forms of the structure, and all geometric or conformational isomer forms of the structure. For example, the R and S configurations of each stereocenter are considered to be part of the present disclosure. Therefore, the single stereochemical isomers of the compound provided, as well as enantiomers, diastereomers, and geometric (or conformational) mixtures are within the scope of the present disclosure. For example, in some cases, the compound provided illustrates one or more stereoisomers of the compound, and unless otherwise stated, represents each stereoisomer individually and / or as a mixture. Unless otherwise stated, all tautomeric forms of the compound provided are within the scope of the present disclosure.

[0313] Unless otherwise indicated, structures depicted herein are intended to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the disclosed structures including replacement of hydrogen by deuterium or tritium or replacement of carbon by 13C-enriched carbon or 14C-enriched carbon are within the scope of the present disclosure.

[0314] About: The term "about" when used herein to refer to a value refers to values ​​that are similar to the referenced value in context. Generally, one skilled in the art familiar with the context will understand the relative degree of variation encompassed by "about" in that context. For example, in some embodiments, the term "about" can encompass a range of values ​​that are within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1% or less of the referenced value.

[0315] Agent: As used herein, the term "agent" can refer to a physical entity. In some embodiments, an agent can be characterized by specific characteristics and / or effects. For example, as used herein, the term "therapeutic agent" refers to a physical entity that has a therapeutic effect and / or elicits a desired biological and / or pharmacological effect. In some embodiments, an agent can be a compound, molecule, or entity of any chemical class, including, for example, small molecules, polypeptides, nucleic acids, sugars, lipids, metals, or combinations or complexes thereof.

[0316] Aliphatic: The term "aliphatic" refers to a straight chain (i.e., unbranched) or branched, substituted or unsubstituted hydrocarbon chain that is fully saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more unsaturated units but is not aromatic (also referred to herein as "cycloaliphatic"), having a single point of attachment or more than one point of attachment to the rest of the molecule. Unless otherwise specified, an aliphatic group contains 1-12 aliphatic carbon atoms. In some embodiments, an aliphatic group contains 1-6 aliphatic carbon atoms (e.g., C 1-6 In some embodiments, aliphatic groups contain 1-5 aliphatic carbon atoms (e.g., C 1-5 In other embodiments, aliphatic groups contain 1-4 aliphatic carbon atoms (e.g., C 1-4 In other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms (e.g., C 1-3 ), and in other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms (e.g., C 1-2 Suitable aliphatic groups include, but are not limited to, straight or branched, substituted or unsubstituted alkyl, alkenyl or alkynyl groups and hybrids thereof. Preferred aliphatic groups are C 1-6 alkyl.

[0317] Alkyl: The term "alkyl," used alone or as part of a larger moiety, refers to a group having (unless otherwise indicated) 1-12, 1-10, 1-8, 1-6, 1-4, 1-3, or 1-2 carbon atoms (e.g., C 1-12 、C 1-10 、C 1-8 、C 1-6 、C 1-4 、C 1-3 or C 1-2 ) is a saturated, optionally substituted, straight or branched chain hydrocarbon group. Exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl and heptyl.

[0318] Alkylene: The term "alkylene" refers to a divalent alkyl group. In some embodiments, an "alkylene" is a divalent straight or branched chain alkyl group. In some embodiments, an "alkylene chain" is a polymethylene group, i.e., -(CH2)n -, wherein n is a positive integer, such as 1 to 6, 1 to 4, 1 to 3, 1 to 2 or 2 to 3. Optionally substituted alkylene chains are polymethylenes in which one or more methylene hydrogen atoms are optionally replaced by substituents. Suitable substituents include those described below for substituted aliphatic groups, and also include those described in the specification herein. It should be understood that two substituents of the alkylene group can be combined to form a ring system. In certain embodiments, the two substituents can be combined to form a 3- to 7-membered ring. The substituents may be on the same or different atoms. The suffix "-ene" or "-enyl" when attached to certain groups herein is intended to refer to the difunctional portion of the group. For example, "-ene" or "-enyl" when attached to "cyclopropyl" becomes "cyclopropene" or "cyclopropenyl" and is intended to refer to a difunctional cyclopropyl group, such as

[0319] Alkenyl: The term "alkenyl," used alone or as part of a larger moiety, refers to a group having at least one double bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms (e.g., C 2-12 、C 2-10 、C 2-8 、C 2-6 、C 2-4 or C 2-3 ) is an optionally substituted straight or branched chain or cyclic hydrocarbon group. Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl. The term "cycloalkenyl" refers to an optionally substituted non-aromatic monocyclic or polycyclic ring system containing at least one carbon-carbon double bond and having from about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkenyl rings include cyclopentenyl, cyclohexenyl, and cycloheptenyl.

[0320] Alkynyl: The term "alkynyl," used alone or as part of a larger moiety, refers to a radical having at least one triple bond and having (unless otherwise specified) 2-12, 2-10, 2-8, 2-6, 2-4, or 2-3 carbon atoms (e.g., C 2-12 、C 2-10 、C 2-8 、C 2-6 、C 2-4 or C 2-3 ) is an optionally substituted straight or branched chain hydrocarbon group. Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.

[0321] Amino Acid: In the broadest sense, as used herein, the term "amino acid" refers to a compound and / or substance that can be, is, or has been incorporated into a polypeptide chain, for example, by forming one or more peptide bonds. In some embodiments, an amino acid has the general structure H2N-C(H)(R)-COOH. In some embodiments, the amino acid is a naturally occurring amino acid. In some embodiments, the amino acid is a non-natural amino acid; in some embodiments, the amino acid is a D-amino acid; in some embodiments, the amino acid is an L-amino acid. "Standard amino acid" refers to any of the twenty standard L-amino acids commonly found in naturally occurring peptides. "Non-standard amino acid" refers to any amino acid other than the standard amino acids, whether synthetically prepared or obtained from a natural source. In some embodiments, the amino acids in a polypeptide (including the carboxyl and / or amino terminal amino acids) may contain structural modifications compared to the general structure described above. For example, in some embodiments, an amino acid may be modified compared to the general structure by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of the amino group, the carboxylic acid group, one or more protons, and / or hydroxyl groups), amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution. In some embodiments, such modifications may, for example, alter the circulating half-life of a polypeptide comprising the modified amino acid, compared to a polypeptide comprising an otherwise identical unmodified amino acid. In some embodiments, such modifications do not significantly alter the relevant activity of a polypeptide comprising the modified amino acid, compared to a polypeptide comprising an otherwise identical unmodified amino acid. As will be clear from the context, in some embodiments, the term "amino acid" may be used to refer to a free amino acid; in some embodiments, it may be used to refer to an amino acid residue of a polypeptide.

[0322] Antibody agent: As used herein, the term "antibody agent" refers to any polypeptide or polypeptide complex comprising immunoglobulin structural elements sufficient to confer specific binding to a specific antigen. Exemplary antibody agents include, but are not limited to, monoclonal antibodies or polyclonal antibodies. In some embodiments, an antibody agent may include one or more constant region sequences that are characteristic of mouse, rabbit, primate, or human antibodies. In some embodiments, an antibody agent may include one or more humanized, primatized, chimeric, and other sequence elements, as known in the art. In some embodiments, the term "antibody agent" is used to refer to one or more constructs or forms known or developed in the art for utilizing antibody structure and functional characteristics in alternative presentations. For example, in some embodiments, the antibody agent used in accordance with the present disclosure is a form selected from, but not limited to, complete IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies (e.g., etc.); CrossMab (e.g., CrossMab CH1-CL ;CrossMab CH1-CLcv ; Bispecific CrossMab with a knob-and-holeCH1-CL ); antibody fragments, such as Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments and isolated complementarity determining regions (CDRs) or collections thereof; single-chain Fv (scFv); scFv-Fc fusions; polypeptide-Fc fusions; single domain antibodies (e.g., shark single domain antibodies, such as IgNAR or fragments thereof); camelid antibodies; masked antibodies (e.g., ); small modular immunopharmaceuticals ("SMIPsTM"); single-chain or tandem bifunctional antibodies VHH; small antibodies; Ankyrin repeat protein or DART;TCR-like antibody; MicroProteins; as well as In some embodiments, the chain and / or fragment of such antibodies and fragments can be used in combination, for example, scFv-Fc arms are used in combination with conventional antibody arms. In some embodiments, the antibody agent is a broad-spectrum neutralizing antibody agent (for example, a broad-spectrum neutralizing antibody (bNAb)). "Broad-spectrum neutralizing antibody agent" is an antibody agent that can neutralize two or more genetic variants (for example, strains) of a virus (for example, HIV). In some embodiments, the antibody may lack the covalent modification (for example, the connection of a polysaccharide) that it would have in the case of natural production. In some embodiments, the antibody may contain a covalent modification (for example, the connection of a polysaccharide, a payload (for example, a detectable portion, a therapeutic portion, a catalytic portion, etc.) or other side groups (for example, polyethylene glycol, etc.). In many embodiments, the antibody agent is or comprises a polypeptide having an amino acid sequence comprising one or more structural elements recognized by those skilled in the art as complementarity determining regions (CDRs); in some embodiments, the antibody agent is or comprises a polypeptide having an amino acid sequence comprising at least one CDR (e.g., at least one heavy chain CDR and / or at least one light chain CDR) that is substantially identical to a CDR found in a reference antibody. In some embodiments, the included CDR is substantially identical to the reference CDR because it is identical in sequence or contains 1-5 amino acid substitutions compared to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because it exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because it exhibits at least 95%, 96%, 97%, 98%, 99% or 100% sequence identity to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because at least one amino acid is deleted, added or substituted within the included CDR as compared to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the amino acid sequence of the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because 1-5 amino acids are deleted, added or substituted within the included CDR as compared to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the amino acid sequence of the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR because at least one amino acid is substituted within the included CDR as compared to the reference CDR, but the included CDR has an amino acid sequence that is otherwise identical to the amino acid sequence of the reference CDR.In some embodiments, the included CDRs are substantially identical to the reference CDRs because 1-5 amino acids are deleted, added, or substituted within the included CDRs as compared to the reference CDRs, but the included CDRs have amino acid sequences that are otherwise identical to the reference CDRs. In some embodiments, the antibody agent is or comprises a polypeptide whose amino acid sequence includes structural elements recognized by those skilled in the art as immunoglobulin variable domains. In some embodiments, the antibody agent is a polypeptide protein having a binding domain that is homologous or largely homologous to an immunoglobulin binding domain.

[0323] Aryl: The term "aryl" refers to monocyclic and bicyclic ring systems having a total of six to fourteen ring members (e.g., C6-C14), wherein at least one ring in the system is aromatic and wherein each ring in the system contains three to seven ring members. In some embodiments, "aryl" contains between six and twelve total ring members (e.g., C6-C12). The term "aryl" can be used interchangeably with the term "aromatic ring". In certain embodiments, "aryl" refers to an aromatic ring system that can carry one or more substituents, including but not limited to phenyl, biphenyl, naphthyl, anthracenyl, and the like. Unless otherwise specified, "aryl" is a hydrocarbon. In some embodiments, an "aryl" ring system is an aromatic ring (e.g., phenyl) fused to a non-aromatic ring (e.g., cycloalkyl).

[0324] Examples of fused aromatic rings include

[0325] Correlation: As used herein, two events or entities are "associated" with each other if the presence, level, extent, type, and / or form of one event or entity is correlated with the presence, level, extent, type, and / or form of another event or entity. For example, if the presence, level, and / or form of a particular entity (e.g., a polypeptide, genetic signature, metabolite, microorganism, etc.) is correlated with the incidence, susceptibility, severity, stage, etc. of a particular disease, disorder, or condition (e.g., among related populations), it is considered to be associated with the disease, disorder, or condition. In some embodiments, two or more entities are physically "associated" with each other if they interact, directly or indirectly, such that they are brought into physical proximity and / or remain in physical proximity to each other. In some embodiments, two or more entities that are physically associated with each other are covalently linked to each other; in some embodiments, two or more entities that are physically associated with each other are not covalently linked to each other, but are non-covalently associated, for example, by means of hydrogen bonds, van der Waals interactions, hydrophobic interactions, magnetic forces, and combinations thereof.

[0326] Co-administration: As used herein, the term "co-administration" refers to the use of a composition (e.g., a pharmaceutical composition) as described herein and one or more additional therapeutic agents. In some embodiments, one or more additional therapeutic agents comprise at least one polyribonucleotide encoding another antibody agent (e.g., an anti-HIV antigen antibody agent). The combined use of a composition (e.g., a pharmaceutical composition) as described herein and an additional therapeutic agent can be performed simultaneously or separately (e.g., sequentially in any order). In some embodiments, a composition (e.g., a pharmaceutical composition) as described herein and an additional therapeutic agent can be combined in a pharmaceutically acceptable excipient, or they can be placed in a separate excipient and delivered to a target cell or administered to a subject at different times. As long as the composition (e.g., a pharmaceutical composition) and an additional therapeutic agent as described herein are delivered or administered close enough in time so that the biological effects produced by each on the target cell or the subject being treated overlap at least some time, each of these situations is considered to be within the meaning of "co-administration" or "combination."

[0327] Combination therapy: As used herein, the term "combination therapy" refers to those situations in which a subject is exposed to two or more treatment regimens (e.g., two or more therapeutic agents (e.g., two or more antibody agents)) at the same time. In some embodiments, two or more regimens can be administered simultaneously; in some embodiments, such regimens can be administered sequentially (e.g., all "doses" of the first regimen are administered, followed by any doses of the second regimen); in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, the administration of the combination therapy may involve administering one or more agents or modalities to a subject who is receiving the other agents or modalities in the combination. For clarity, combination therapy does not require that the individual agents be administered together in a single composition (or not even necessarily at the same time), but in some embodiments, two or more agents or active portions thereof can be administered together in a combination composition. In some embodiments, the combination therapy comprises polyribonucleotides encoding two or more antibody agents (e.g., anti-HIV antibody agents).

[0328] Comparable: As used herein, the term "comparable" refers to two or more agents, entities, situations, sets of conditions, etc. that may not be identical to one another, but are similar enough to allow comparisons to be made between them so that one skilled in the art will understand that conclusions can be reasonably drawn based on the observed differences or similarities. In some embodiments, a set of comparable conditions, situations, individuals, or populations is characterized by a plurality of substantially identical features and one or a small amount of variation. One of ordinary skill in the art will understand that, in the context, in any given case, what degree of identity is required for a set of two or more such agents, entities, situations, conditions, etc. to be considered comparable. For example, one of ordinary skill in the art will understand that when a set of situations, individuals, or populations is characterized by a sufficient number and type of substantially identical features, they are comparable to one another to ensure that the conclusion is reasonable, i.e., the differences in the results or observed phenomena obtained under or with different sets of situations, individuals, or populations are caused by or indicate variations in those features that have varied.

[0329] Corresponding to: As used herein, the term "corresponding to" refers to a relationship between two or more entities. For example, the term "corresponding to" can be used to indicate the position / identity of a structural element in a compound or composition relative to another compound or composition (e.g., relative to an appropriate reference compound or composition). For example, in some embodiments, a monomer residue in a polymer (e.g., an amino acid residue in a polypeptide or a nucleic acid residue in a polynucleotide) can be identified as a residue in an appropriate reference polymer. For example, one of ordinary skill in the art will appreciate that, for simplicity, residues in polypeptides are often designated using a canonical numbering system based on a reference to a related polypeptide, such that, for example, an amino acid that "corresponds to" a residue at position 190 is not actually necessarily the 190th amino acid in a particular amino acid chain, but rather corresponds to the residue found at 190 in a reference polypeptide; one of ordinary skill in the art will readily understand how to identify a "corresponding" amino acid. For example, those skilled in the art will be aware of various sequence alignment strategies, including software programs such as BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH / GLSEARCH, Genoogle, HMMER, HHpred / HHsearch, IDF, Infernal, KLAST, USEARCH, parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE, which can be used, for example, to identify "corresponding" residues in polypeptides and / or nucleic acids according to the present disclosure. Those skilled in the art will also appreciate that, in some cases, the term "corresponding to" can be used to describe an event or entity that shares a relevant similarity with another event or entity (e.g., an appropriate reference event or entity). As just one example, a gene or protein in one organism may be described as "corresponding to" a gene or protein from another organism to indicate that, in some embodiments, it plays a similar role or performs a similar function and / or that it exhibits a particular degree of sequence identity or homology, or shares particular characteristic sequence elements.

[0330] Cycloaliphatic: As used herein, the term "cycloaliphatic" refers to a monocyclic C-1-cyclic alkyl radical that is fully saturated or contains one or more unsaturated units but is not aromatic and has a single point of attachment or more than one point of attachment to the rest of the molecule. 3-8 Hydrocarbon or bicyclic C 6-10 hydrocarbon.

[0331] Cycloalkyl: As used herein, the term "cycloalkyl" refers to an optionally substituted, saturated, monocyclic or polycyclic ring system of about 3 to about 10 ring carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0332] Derived: In the context of an amino acid sequence (peptide or polypeptide) that is "derived from" a specified amino acid sequence (peptide or polypeptide), it refers to a structural analog of the specified amino acid sequence. In some embodiments, the amino acid sequence derived from a specific amino acid sequence has an amino acid sequence that is identical, substantially identical, or homologous to the specific sequence or a fragment thereof. The amino acid sequence derived from a specific amino acid sequence may be a variant of the specific sequence or a fragment thereof. For example, the antibody agent used in accordance with the present disclosure may include amino acid sequences (e.g., CDRs, variable domains, constant domains, etc.) derived from other antibodies (e.g., naturally occurring antibodies).

[0333] Detection: The term "detection" is used broadly herein to include appropriate means for determining the presence or absence of an entity of interest in a sample or any form of measurement of an entity of interest. Thus, "detection" can include determining, measuring, evaluating or analyzing the presence or absence, level, amount and / or position of an entity of interest. Includes quantitative and qualitative determination, measurement or evaluation, including semi-quantitative. Such determination, measurement or evaluation can be relative (e.g., when the entity of interest is detected relative to a control reference), or absolute. Thus, the term "quantitative" can refer to absolute or relative quantification when used in the context of quantitatively measuring an entity of interest. Absolute quantification can be achieved by associating the level of the detected entity of interest with a known control standard (e.g., by generating a standard curve). Alternatively, relative quantification can be achieved by comparing the level or amount detected between two or more different entities of interest to provide relative quantification of each of the two or more different entities of interest (i.e., relative to each other).

[0334] Dosing regimen: Those skilled in the art will appreciate that the term "dosing regimen" (or "therapeutic regimen") can be used to refer to a group of unit doses (usually more than one) that are administered individually to a subject, usually at intervals of time. In some embodiments, a given therapeutic agent has a recommended dosing regimen that may involve one or more doses.

[0335] Encoding: As used herein, the term "encode / encoding" refers to the sequence information of a first molecule, which guides the production of a second molecule having a determined nucleotide sequence (e.g., a polyribonucleotide) or a determined amino acid sequence. For example, a DNA molecule can encode an RNA molecule (e.g., through a transcription process involving a DNA-dependent RNA polymerase). An RNA molecule can encode a polypeptide (e.g., through a translation process). Therefore, if the transcription and translation of the RNA corresponding to a gene produces a polypeptide in a cell or other biological system, the gene, cDNA, or RNA molecule encodes the polypeptide. In some embodiments, the coding region of a polyribonucleotide encoding a target antigen refers to a coding chain whose nucleotide sequence is identical to the polyribonucleotide sequence of such a target antigen. In some embodiments, the coding region of a polyribonucleotide encoding a target antigen refers to a non-coding chain of such a target antigen, which can be used as a transcription template for a gene or cDNA.

[0336] Engineered: Generally, the term "engineered" refers to aspects that have been manipulated by the hand of man. For example, a polynucleotide is considered "engineered" when two or more sequences that are not linked together in nature are directly linked to each other in an engineered polynucleotide by human manipulation and / or when specific residues in a polynucleotide are non-naturally occurring and / or are caused by the action of man to be linked to entities or moieties to which they are not linked in nature.

[0337] Epitope: As used herein, the term "epitope" refers to a portion that is specifically recognized by an immunoglobulin (e.g., an antibody or receptor) binding component. For example, an epitope can be recognized by a T cell, a B cell, or an antibody. In some embodiments, an epitope comprises a plurality of chemical atoms or groups on an antigen. In some embodiments, when the antigen adopts a related three-dimensional conformation, such chemical atoms or groups are surface exposed. In some embodiments, when the antigen adopts such a conformation, such chemical atoms or groups are physically close to each other in space. In some embodiments, when the antigen adopts an alternative conformation (e.g., linearized), at least some of such chemical atoms or groups are physically separated from each other. Therefore, in some embodiments, the epitope of an antigen may include a continuous or discontinuous portion of the antigen. In some embodiments, an epitope is or comprises a T cell epitope. In some embodiments, an epitope may have a length of about 5 to about 30 amino acids, or about 10 to about 25 amino acids, or about 5 to about 15 amino acids, or about 5 to 12 amino acids, or about 6 to about 9 amino acids.

[0338] Expression: As used herein, the term "expression" of a nucleic acid sequence refers to the production of a gene product from the nucleic acid sequence. In some embodiments, the gene product can be a transcript, such as a polyribonucleotide as provided herein. In some embodiments, the gene product can be a polypeptide. In some embodiments, expression of a nucleic acid sequence involves one or more of the following: (1) production of an RNA template from the DNA sequence (e.g., by transcription); (2) processing of the RNA transcript (e.g., by splicing, editing, etc.); (3) translation of the RNA into a polypeptide or protein; and / or (4) post-translational modification of the polypeptide or protein.

[0339] Heteroaliphatic: As used herein, the term "heteroaliphatic" or "heteroaliphatic group" means an optionally substituted hydrocarbon moiety having from one to five heteroatoms in addition to carbon atoms, which may be straight chain (i.e., unbranched), branched, or cyclic ("heterocycle") and may be fully saturated, or may contain one or more unsaturated units but is not aromatic. The term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur and any quaternized form of basic nitrogen. The term "nitrogen" also includes substituted nitrogen. Unless otherwise specified, heteroaliphatic groups contain 1-10 carbon atoms, of which 1-3 carbon atoms are optionally and independently replaced by heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, heteroaliphatic groups contain 1-4 carbon atoms, of which 1-2 carbon atoms are optionally and independently replaced by heteroatoms selected from oxygen, nitrogen, and sulfur. In other embodiments, heteroaliphatic groups contain 1-3 carbon atoms, of which 1 carbon atom is optionally and independently replaced by a heteroatom selected from oxygen, nitrogen, and sulfur. Suitable heteroaliphatic groups include, but are not limited to, straight or branched chain heteroalkyl, heteroalkenyl, and heteroalkynyl groups. For example, heteroaliphatic groups of 1 to 10 atoms include the following exemplary groups: O-CH3, -CH2-O-CH3, -O-CH2-CH2-O-CH2-CH2-O-CH3, etc.

[0340] Heteroaryl: The terms "heteroaryl" and "heteroar-" used alone or as part of a larger moiety (e.g., "heteroaralkyl" or "heteroaralkoxy") refer to monocyclic or bicyclic groups having 5 to 10 ring atoms (e.g., a 5- to 6-membered monocyclic heteroaryl or a 9- to 10-membered bicyclic heteroaryl); having 6, 10, or 14 pi electrons shared in the cyclic array; and having one to five heteroatoms in addition to carbon atoms. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridinyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[1,2-a]pyrimidinyl, imidazo[1,2-a]pyridinyl, imidazo[4,5-b]pyridinyl, imidazo[4,5-c]pyridinyl, pyrrolopyridinyl, pyrrolopyrazinyl, thienopyrimidinyl, triazolopyridinyl, and benzisoxazolyl. As used herein, the terms "heteroaryl" and "heteroar-" also include groups in which a heteroaromatic ring is fused to one or more aryl, cycloaliphatic, or heterocyclyl rings, where the radical or point of attachment is on the heteroaromatic ring (i.e., bicyclic heteroaromatic rings having from 1 to 3 heteroatoms). Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzotriazolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolinyl, isoquinolinyl, cinnolinyl, phthalazinyl, quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl, phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl, pyrido[2,3-b]-1,4-oxazin-3(4H)-one, 4H-thieno[3,2-b]pyrrole, and benzisoxazolyl. The term "heteroaryl" may be used interchangeably with the terms "heteroaromatic ring," "heteroaryl," or "heteroaromatic," any of which includes optionally substituted rings.

[0341] Heteroatom: As used herein, the term "heteroatom" refers to nitrogen, oxygen, or sulfur, and includes any oxidized form of nitrogen or sulfur, and any quaternized form of a basic nitrogen.

[0342] Heterocycle: As used herein, the terms "heterocycle", "heterocyclyl", "heterocyclic radical" and "heterocyclic ring" are used interchangeably and refer to a stable 3- to 8-membered monocyclic, 6- to 10-membered bicyclic or 10- to 16-membered polycyclic heterocyclic moiety that is saturated or partially unsaturated and has one or more (such as one to four) heteroatoms as defined above in addition to carbon atoms. The term "nitrogen" when used to refer to the ring atoms of a heterocycle includes substituted nitrogen. For example, in a saturated or partially unsaturated ring having 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, nitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) or NR + (as in the pyrrolidinyl of N- substitution).Heterocycle can be connected to its side group at any heteroatom or carbon atom, so as to produce stable structure, and any ring atom can be optionally substituted.The example of such saturated or partially unsaturated heterocyclic group includes but is not limited to azetidinyl (azetidinyl), oxetanyl (oxetanyl), tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolane, diazepine, oxazepine, sulphuretrazolyl, morpholinyl and thiomorpholinyl.Heterocyclic group can be monocycle, bicycle, tricycle or polycycle, preferably monocycle, bicycle or tricycle, more preferably monocycle or bicycle.Bicyclic heterocycle also includes the group in which heterocycle is fused with one or more aromatic rings. Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl and tetrahydroquinolinyl. The bicyclic heterocycle can also be a spiro ring system (e.g., a 7- to 11-membered spiro ring fused heterocycle having one or more heteroatoms (e.g., one, two, three or four heteroatoms) as defined above in addition to carbon atoms). The bicyclic heterocycle can also be a bridged ring system (e.g., a 7- to 11-membered bridged heterocycle having one, two or three bridging atoms).

[0343] Homology: As used herein, the terms "homology" or "homologs" refer to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "homologous" to each other if their sequences are at least 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% identical. In some embodiments, if the sequences of polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95% or 99% similar (e.g., containing residues related in chemical properties at corresponding positions), they are considered to be "homologous" to each other. For example, as is well known to those skilled in the art, certain amino acids are typically classified as "hydrophobic" or "hydrophilic" amino acids that are similar to each other, and / or have "polar" or "non-polar" side chains. Substituting one amino acid for another amino acid of the same type is generally considered a "homologous" substitution.

[0344] Identity: As used herein, the term "identity" refers to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. In some embodiments, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "substantially identical" to each other if their sequences are at least 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99% identical. The calculation of the percent identity of two nucleic acid or polypeptide sequences can be performed, for example, by aligning the two sequences for the purpose of optimal comparison (e.g., gaps can be introduced in one or both of the first and second sequences for optimal alignment, and non-identical sequences can be ignored for the purpose of comparison). In certain embodiments, the length of the sequence compared for the purpose of comparison is at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or substantially 100% of the length of the reference sequence. The nucleotides at the corresponding positions are then compared. When the position in the first sequence is occupied by the same residue (e.g., nucleotide or amino acid) as the corresponding position in the second sequence, the molecules are identical at the position. Taking into account the number of gaps and the length of each gap, the percent identity between the two sequences is a function of the number of identical positions shared by the sequences, which gaps need to be introduced for the optimal comparison of the two sequences. Mathematical algorithms can be used to complete the comparison of the sequences between the two sequences and the determination of percent identity. For example, the algorithm of Meyers and Miller, 1989, incorporated into the ALIGN program (version 2.0), can be used to determine the percent identity between two nucleotide sequences. In some exemplary embodiments, nucleic acid sequence comparisons performed with the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Alternatively, the GAP program in the GCG software package can be used to determine the percent identity between two nucleotide sequences using the NWSgapdna.CMP matrix.

[0345] Increased, induced, or decreased: As used herein, these terms or grammatically equivalent comparative terms indicate a value relative to a comparable reference measurement. For example, in some embodiments, an assessment obtained with a provided composition (e.g., a pharmaceutical composition) may be "increased" relative to an assessment obtained with a comparable reference composition. Alternatively or additionally, in some embodiments, an assessment obtained in a subject may be "increased" relative to an assessment obtained under different conditions in the same subject (e.g., before or after an event; or with or without an event, such as administration of a composition as described herein (e.g., a pharmaceutical composition) or in a different comparable subject (e.g., in a comparable subject different from the subject of interest, the subject of interest having been previously exposed to conditions in which, for example, the absence of administration of a composition as described herein (e.g., a pharmaceutical composition)). In some embodiments, comparative terms refer to statistically relevant differences (e.g., prevalence and / or magnitude sufficient to achieve statistical relevance). Those skilled in the art will recognize or will readily be able to provide a comparative term for a given subject. In some embodiments, the term "reduced" or equivalent terms refers to a decrease in the level of an assessed value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75% or more compared to a comparable reference. In some embodiments, the term "reduced" or equivalent terms refers to complete or substantially complete inhibition, i.e., reduction to zero or substantially to zero. In some embodiments, the term "increased" or "induced" refers to an increase in the level of an assessed value by at least 10%, at least 20%, at least 30%, at least 40%, at least 50%, at least 80%, at least 100%, at least 200%, at least 500% or more compared to a comparable reference.

[0346] In sequence: As used herein with respect to polynucleotides or polyribonucleotides, "in sequence" refers to the order of features along the polynucleotide or polyribonucleotide from 5' to 3'. As used herein with respect to polypeptides, "in sequence" refers to the order of features along the polypeptide moving from the feature closest to the N-terminus to the feature closest to the C-terminus. "In sequence" does not mean that additional features cannot be present in the listed features. For example, if features A, B, and C of a polynucleotide are described herein as "in sequence as feature A, feature B, and feature C," such description does not exclude, for example, that feature D is located between features A and B.

[0347] Ionizable: The term "ionizable" refers to a compound or group or atom that is charged at a certain pH. In the context of ionizable amino lipids, such lipids or their functional groups or atoms carry a positive charge at a certain pH. In some embodiments, ionizable amino lipids are positively charged at acidic pH. In some embodiments, ionizable amino lipids are mainly neutral at physiological pH (e.g., about 7.0-7.4 in some embodiments), but become positively charged at lower pH values. In some embodiments, ionizable amino lipids can have a pKa in the range of about 5 to about 7.

[0348] Isolated: The term "isolated" means altered or removed from its native state. For example, a nucleic acid or peptide naturally occurring in a living animal is not "isolated," but the same nucleic acid or peptide partially or completely separated from the coexisting materials of its native state is "isolated." An isolated nucleic acid or protein can exist in a substantially purified form or can exist in a non-native environment (e.g., a host cell).

[0349] Lipid: As used herein, the terms "lipid" and "lipid-like material" are broadly defined as molecules comprising one or more hydrophobic portions or groups and optionally also comprising one or more hydrophilic portions or groups. Molecules comprising a hydrophobic portion and a hydrophilic portion are also commonly referred to as amphiphilic molecules.

[0350] RNA lipid nanoparticles: as used herein, the term "RNA lipid nanoparticles" refers to nanoparticles comprising at least one lipid and RNA molecules, for example, one or more polyribonucleotides as provided herein. In some embodiments, the RNA lipid nanoparticles comprise at least one cationic amino lipid. In some embodiments, the RNA lipid nanoparticles comprise at least one cationic amino lipid, at least one helper lipid, and at least one polymer-conjugated lipid (e.g., PEG-conjugated lipid). In various embodiments, the RNA lipid nanoparticles as described herein can have an average size (e.g., Z average) of about 100 nm to 1000 nm, or about 200 nm to 900 nm, or about 200 nm to 800 nm, or about 250 nm to about 700 nm. In some embodiments of the present disclosure, the RNA lipid nanoparticles can have a particle diameter (e.g., Z mean values) of about 30 nm to about 200 nm, or about 30 nm to about 150 nm, about 40 nm to about 150 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, about 70 nm to about 100 nm, about 80 nm to about 100 nm, about 90 nm to about 100 nm, about 70 to about 90 nm, about 80 nm to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, the average size of the lipid nanoparticles is determined by measuring the average particle diameter. In some embodiments, the RNA lipid nanoparticles can be prepared by mixing lipids with RNA molecules as described herein.

[0351] Neutralization: As used herein, the term "neutralization" refers to an event in which a binding agent (such as an antibody) binds to a biologically active site of a virus (such as a receptor binding protein), thereby inhibiting parasitic infection of a cell. In some embodiments, the term "neutralization" refers to an event in which a binding agent eliminates or significantly reduces the ability of a binding agent to infect a cell.

[0352] Nucleic acid / polynucleotide: As used herein, the term "nucleic acid" refers to a polymer of at least 10 or more nucleotides. In some embodiments, the nucleic acid is or comprises DNA. In some embodiments, the nucleic acid is or comprises RNA. In some embodiments, the nucleic acid is or comprises peptide nucleic acid (PNA). In some embodiments, the nucleic acid is or comprises single-stranded nucleic acid. In some embodiments, the nucleic acid is or comprises double-stranded nucleic acid. In some embodiments, the nucleic acid comprises both single-stranded and double-stranded portions. In some embodiments, the nucleic acid comprises a backbone comprising one or more phosphodiester linkages. In some embodiments, the nucleic acid comprises a backbone comprising both phosphodiester and non-phosphodiester linkages. For example, in some embodiments, the nucleic acid may comprise a backbone comprising one or more phosphorothioate or 5'-N-phosphoramidite linkages and / or one or more peptide bonds, such as in "peptide nucleic acids." In some embodiments, the nucleic acid comprises one or more or all natural residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, the nucleic acid comprises one or more or all non-natural residues. In some embodiments, the non-natural residue comprises a nucleoside analog (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolo-pyrimidine, 3-methyladenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, 6-O-methylguanine, 2-thiocytidine, methylated bases, embedded bases, and combinations thereof). In some embodiments, the non-natural residue comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxyribose, arabinose, and hexose) as compared to those in the natural residue. In some embodiments, nucleic acid has a nucleotide sequence encoding a functional gene product (such as RNA or polypeptide). In some embodiments, nucleic acid has a nucleotide sequence comprising one or more introns. In some embodiments, nucleic acid can be prepared by separation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, such as in vivo or in vitro), replication in a recombinant cell or system, or chemical synthesis.In some embodiments, the nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 1 10, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, 5500, 6000, 6500, 7000, 7500, 800 18,000, 18,500, 19,000, 19,500, or 20,000 residues or nucleotides in length.

[0353] Pharmaceutically effective amount: The term "pharmaceutically effective amount" or "therapeutically effective amount" refers to an amount that, alone or in combination with further doses, achieves a desired response or desired effect. In the case of treating a particular disease (e.g., HIV), the desired response, in some embodiments, involves inhibiting the course of the disease (e.g., HIV). In some embodiments, such inhibition may include slowing the progression of the disease (e.g., HIV) and / or interrupting or reversing the progression of the disease (e.g., HIV). In some embodiments, the desired response in the treatment of a disease (e.g., HIV) may be or include delaying or preventing the onset of a disease (e.g., HIV) or illness (e.g., HIV-related illness). The effective amount of a composition (e.g., a pharmaceutical composition) described herein will depend, for example, on the disease (e.g., HIV) or illness (e.g., HIV-related illness) to be treated, the severity of such disease (e.g., HIV) or illness (e.g., HIV-related illness), the individual parameters of the patient (including, for example, age, physiological condition, body size and weight), the duration of treatment, the type of concomitant therapy (if any), the specific route of administration, and similar factors. Thus, the dosage of the compositions described herein (e.g., pharmaceutical compositions) may depend on a variety of such parameters. In cases where the patient's response to the initial dose is insufficient, a higher dose (or an effectively higher dose obtained by a different, more localized route of administration) may be used.

[0354] Polypeptide: As used herein, the term "polypeptide" refers to a polymeric chain of amino acids. In some embodiments, a polypeptide has an amino acid sequence that occurs in nature. In some embodiments, a polypeptide has an amino acid sequence that does not occur in nature. In some embodiments, a polypeptide has an engineered amino acid sequence, meaning that the sequence is designed and / or generated by the hand of man. In some embodiments, a polypeptide may comprise or consist of natural amino acids, non-natural amino acids, or both. In some embodiments, a polypeptide may comprise or consist of only natural amino acids or only non-natural amino acids. In some embodiments, a polypeptide may comprise D-amino acids, L-amino acids, or both. In some embodiments, a polypeptide may comprise only D-amino acids. In some embodiments, a polypeptide may comprise only L-amino acids. In some embodiments, a polypeptide may include one or more side groups or other modifications, such as modifications or attachment to one or more amino acid side chains at the N-terminus, the C-terminus, or any combination thereof. In some embodiments, such side groups or modifications include acetylation, amidation, lipidation, methylation, pegylation, and the like, including combinations thereof. In some embodiments, a polypeptide may be cyclic and / or may include a cyclic portion. In some embodiments, the polypeptide is not cyclic and / or does not contain any cyclic portions. In some embodiments, the polypeptide is linear. In some embodiments, the polypeptide can be or contain a stapled polypeptide. In some embodiments, the term "polypeptide" can be appended to the name, activity, or structure of a reference polypeptide; in such cases, it is used herein to refer to polypeptides that share a related activity or structure and can therefore be considered members of the same class or family of polypeptides. For each such class, the specification provides and / or those skilled in the art will be aware of exemplary polypeptides of known amino acid sequence and / or function within the class; in some embodiments, such exemplary polypeptides are reference polypeptides of the class or family of polypeptides. In some embodiments, members of a class or family of polypeptides exhibit significant sequence homology or identity with a reference polypeptide of the class, share common sequence motifs (e.g., characteristic sequence elements) with a reference polypeptide of the class, and / or share a common activity (in some embodiments at comparable levels or within a specified range); in some embodiments, this is for all polypeptides within the class).For example, in some embodiments, member polypeptides exhibit a degree of overall sequence homology or identity to a reference polypeptide of at least about 30-40%, and typically greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or more, and / or include at least one region (e.g., a conserved region that may be or include characteristic sequence elements in some embodiments) that exhibits very high sequence identity, typically greater than 90% or even 95%, 96%, 97%, 98% or 99%. Such conserved regions typically encompass at least 3-4 and often up to 35 or more amino acids; in some embodiments, a conserved region encompasses at least one stretch of at least 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35 or more contiguous amino acids. In some embodiments, a related polypeptide may comprise or consist of a fragment of a parent polypeptide.

[0355] Prevent: As used herein, the terms "prevent" and "prevention" when used in conjunction with the occurrence of a disease, condition, and / or disorder refer to reducing the risk of developing the disease, condition, and / or disorder and / or delaying the onset of one or more characteristics or symptoms of the disease, condition, or disorder. Prevention can be considered complete when the onset of the disease, condition, or disorder has been delayed for a predetermined period of time.

[0356] Reference: As used herein, the term "reference" describes a standard or control to which a comparison is made. For example, in some embodiments, an agent, animal, individual, population, sample, sequence, or value of interest is compared to a reference or control agent, animal, individual, population, sample, sequence, or value. In some embodiments, a reference or control is tested and / or determined substantially concurrently with the test or determination of interest. In some embodiments, a reference or control is a historical reference or control, optionally embodied in a tangible medium. Typically, as will be understood by those skilled in the art, a reference or control is determined or characterized under conditions or circumstances comparable to those being evaluated. Those skilled in the art will understand when there is sufficient similarity to justify reliance on and / or comparison to a particular possible reference or control.

[0357] Ribonucleic acid (RNA) or polyribonucleotide: As used herein, the terms "ribonucleic acid," "RNA," or "polyribonucleotide" refer to polymers of ribonucleotides. In some embodiments, RNA is single-stranded. In some embodiments, RNA is double-stranded. In some embodiments, RNA comprises both single-stranded and double-stranded portions. In some embodiments, RNA may comprise a backbone structure as described above in the definition of "nucleic acid / polynucleotide." RNA may be a regulatory RNA (e.g., siRNA, microRNA, etc.) or a messenger RNA (mRNA). In some embodiments, RNA is mRNA. In some embodiments in which RNA is mRNA, RNA typically comprises a poly(A) region at its 3' end. In some embodiments in which RNA is mRNA, RNA typically comprises a cap structure recognized in the art at its 5' end, for example, for recognizing mRNA and attaching it to ribosomes to initiate translation. In some embodiments, RNA is synthetic RNA. Synthetic RNA includes RNA synthesized in vitro (e.g., by enzymatic synthesis methods and / or by chemical synthesis methods).

[0358] Ribonucleotide: As used herein, the term "ribonucleotide" encompasses unmodified ribonucleotides and modified ribonucleotides. As used herein, unmodified ribonucleotides include the purine bases adenine (A) and guanine (G) and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides may include one or more modifications, including but not limited to, for example (a) end modifications, such as 5' end modifications (e.g., phosphorylation, dephosphorylation, conjugation, reverse linkage, etc.), 3' end modifications (e.g., conjugation, reverse linkage, etc.), (b) base modifications, such as replacement with modified bases, stabilized bases, destabilized bases, or bases that pair with an expanded partner library or conjugated bases, (c) sugar modifications (e.g., at the 2' position or the 4' position) or replacement of sugars, and (d) internucleoside linkage modifications, including modification or replacement of phosphodiester linkages. The term "ribonucleotide" also encompasses ribonucleotide triphosphates, including modified and unmodified ribonucleotide triphosphates.

[0359] Risk: It will be understood from the context that the "risk" of a disease, condition and / or illness refers to the likelihood that a particular individual will develop the disease, condition and / or illness. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is expressed as a risk relative to the risk associated with a reference sample or reference sample group. In some embodiments, a reference sample or reference sample group has a risk of a known disease, condition, illness and / or event. In some embodiments, a reference sample or reference sample group is from an individual comparable to a particular individual. In some embodiments, risk can reflect one or more genetic attributes, such as which may make an individual susceptible to (or not susceptible to) a particular disease, condition and / or illness. In some embodiments, risk can reflect one or more epigenetic events or attributes and / or one or more lifestyle or environmental events or attributes.

[0360] Selectivity or specificity: The term "selectivity" or "specificity" when used herein to refer to an agent with activity is understood by those skilled in the art to mean that the agent will distinguish between potential target entities, states, or cells. For example, in some embodiments, an agent is considered to "specifically" bind to its target if it preferentially binds to its target in the presence of one or more competing alternative targets. In many embodiments, specific interactions depend on the presence of specific structural features (e.g., epitopes, clefts, binding sites) of the target entity. It should be understood that specificity does not need to be absolute. In some embodiments, specificity can be assessed relative to the specificity of the target binding portion for one or more other potential target entities (e.g., competitors). In some embodiments, specificity is assessed relative to the specificity of a reference specific binding portion. In some embodiments, specificity is assessed relative to the specificity of a reference non-specific binding portion.

[0361] Substituted or optionally substituted: As described herein, compounds of the invention may contain "optionally substituted" moieties. In general, the term "substituted," whether preceded by the term "optionally" or not, means that one or more hydrogen atoms of the designated moiety are replaced with a suitable substituent. "Substituted" applies to one or more hydrogen atoms present in the structure, whether explicitly or implicitly (e.g., At least and At least Unless otherwise noted, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and when one or more positions in any given structure can be substituted by one or more substituents selected from a specified group, the substituents at each position may be the same or different. The combination of substituents contemplated by the present invention is preferably such that a stable or chemically feasible compound is formed. As used herein, the term "stable" refers to a compound that is substantially unchanged when subjected to conditions allowing the manufacture, detection, and, in certain embodiments, the recovery, purification, and use of one or more purposes provided herein for the compound. The group described as "substituted" preferably has between 1 and 4 substituents, more preferably 1 or 2 substituents. The group described as "optionally substituted" may be unsubstituted or "substituted" as described above.

[0362] Suitable monovalent substituents on the substitutable carbon atoms of an "optionally substituted" group are independently halogen; -(CH2) 0-4 R o ; -(CH2) 0-4 OR o ;-O(CH2) 0-4 R o 、-O-(CH2) 0-4 C(O)OR o ; -(CH2) 0-4 CH(OR o )2;-(CH2) 0- 4SR o ; -(CH2) 0-4 Ph, which can be R o Substitution; -(CH2) 0-4 O(CH2) 0-1 Ph, which can be R o Substituted; -CH=CHPh, which can be R o Substitution; -(CH2) 0-4 O(CH2) 0-1 -pyridyl, which may be R o Substitution; -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R o )2;-(CH2) 0-4 N(R o )C(O)R o ;-N(R o )C(S)R o ; -(CH2) 0-4 N(R o )C(O)NR o 2;-N(R o )C(S)NR o2;-(CH2) 0-4 N(R o )C(O)OR o ;-N(R o )N(R o )C(O)R o ;-N(R o )N(R o )C(O)NR o 2;-N(R o )N(R o )C(O)OR o ;-(CH2) 0-4 C(O)R o ;C(S)R o ;-(CH2) 0-4 C(O)OR o ;-(CH2) 0-4 C(O)SR o ;-(CH2) 0-4 C(O)OSiR o 3;-(CH2) 0-4 OC(O)R o ;-OC(O)(CH2) 0-4 SR o ;-(CH2) 0-4 SC(O)R o ;-(CH2) 0-4 C(O)NR o 2;-C(S)NR o 2;-C(S)SR o ;-SC(S)SR o 、-(CH2) 0- 4OC(O)NR o 2;-C(O)N(OR o )R o ;-C(O)C(O)R o ;-C(O)CH2C(O)R o ;-C(NOR o )R o ;-(CH2) 0-4 SSR o ;-(CH2) 0-4 S(O)2R o ;-(CH2) 0-4 S(O)2OR o ;-(CH2) 0-4 OS(O)2R o ;-S(O)2NR o 2;-(CH2) 0-4 S(O)Ro ;-N(R o )S(O)2NR o 2;-N(R o )S(O)2R o ;-N(OR o )R o ;-C(NH)NR o 2;-P(O)2R o ;-P(O)R o 2;-OP(O)R o 2;-OP(O)(OR o )2;SiR o 3;-(C 1-4 linear or branched alkylene)ON(R o )2; or -(C 1-4 Straight or branched alkylene) C(O)ON(R o )2, where each R o may be substituted as defined below and are independently hydrogen, C 1-6 Aliphatic groups, -CH2Ph, -O(CH2) 0- 1Ph, -CH2-(5- to 6-membered heteroaromatic ring) or a 3- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, or notwithstanding the above definitions, two independent occurrences of R o Together with its intervening atoms, it forms a 3- to 12-membered saturated, partially unsaturated or aromatic monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur, which monocyclic or bicyclic ring may be substituted as defined below.

[0363] R o A suitable monovalent substituent on (or by combining two independent occurrences of R o and its intermediate atoms together to form a ring) are independently halogen, -(CH2) 0-2 R l 、-(halogen R l ), -(CH2) 0-2 OH, -(CH2) 0-2 OR l 、-(CH2) 0-2 CH(OR l )2、-O(halogen R l )、-CN、-N3、-(CH2) 0-2 C(O)R l 、-(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR l 、-(CH2) 0-2 SRl 、-(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR l 、-(CH2) 0-2 NR l 2. -NO2, -SiR l 3. -OSiR l 3. -C(O)SR l 、-(C 1-4 linear or branched alkylene)C(O)OR l or -SSR l , where each R l is unsubstituted or, in the case of a "halo" group, substituted only with one or more halogens, and is independently selected from C 1-4 Aliphatic groups, -CH2Ph, -O(CH2) 0-1 Ph or a 3- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. o Suitable divalent substituents on a saturated carbon atom of include =0 and =S.

[0364] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: =0 ("oxo"), =S, =NNR * 2. =NNHC(O)R * 、=NNHC(O)OR * 、=NNHS(O)2R * 、=NR * 、=NOR * 、-O(C(R * 2)) 2- 3O-or-S(C(R * 2)) 2-3 S-, where each independent occurrence of R * is selected from hydrogen, C which may be substituted as defined below 1-6 an aliphatic group or an unsubstituted 5- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur. Suitable divalent substituents bonded to adjacent substitutable carbon atoms of an "optionally substituted" group include: -O(CR * 2) 2-3 O-, where each independent occurrence of R * is selected from hydrogen, C which may be substituted as defined below 1-6 an aliphatic group or an unsubstituted 5-6 membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0365] R* Suitable substituents on the aliphatic group include halogen, -R l 、-(halogen R l ), -OH, -OR l 、-O(halogen R l )、-CN、-C(O)OH、-C(O)OR l 、-NH2、-NHR l 、-NR l 2 or -NO2, where each R l is unsubstituted or, when preceded by "halo", substituted only by one or more halogens, and is independently C 1-4 Aliphatic groups, -CH2Ph, -O(CH2) 0- 1Ph or a 3- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0366] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include Each of these are independently hydrogen, C 1-6 aliphatic, unsubstituted -OPh, or a 3- to 6-membered saturated, partially unsaturated, or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, or, notwithstanding the above definition, two independent occurrences of Together with its intervening atoms, it forms a 3- to 12-membered saturated, partially unsaturated or aromatic monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0367] Suitable substituents on the aliphatic group are independently halogen, -R l 、-(halogen R l ), -OH, -OR l 、-O(halogen R l )、-CN、-C(O)OH、-C(O)OR l 、-NH2、-NHR l 、-NR l 2 or -NO2, where each R l is unsubstituted or, when preceded by "halo", substituted only by one or more halogens, and is independently C 1-4 Aliphatic groups, -CH2Ph, -O(CH2) 0-1 Ph or a 3- to 6-membered saturated, partially unsaturated or aromatic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0368] Subject: As used herein, the term "subject" refers to an organism to which the compositions described herein are to be administered, for example, for experimental, diagnostic, preventive and / or therapeutic purposes. Typical subjects include animals (e.g., mammals, such as mice, rats, rabbits, non-human primates, domestic pets, etc.) and humans. In some embodiments, the subject is a human subject. In some embodiments, the subject suffers from a disease, disorder or illness (e.g., HIV, HIV-related disorders, etc.). In some embodiments, the subject is susceptible to a disease, disorder or illness (e.g., HIV, HIV-related disorders, etc.). In some embodiments, the subject exhibits one or more symptoms or features of a disease, disorder or illness (e.g., HIV, HIV-related disorders, etc.). In some embodiments, the subject exhibits one or more nonspecific symptoms of a disease, disorder or illness (e.g., HIV, HIV-related disorders, etc.). In some embodiments, the subject does not exhibit any symptoms or features of a disease, disorder or illness (e.g., HIV, HIV-related disorders, etc.). In some embodiments, the subject is a person who is susceptible to a disease, condition, or illness (e.g., HIV, HIV-related illnesses, etc.) or has one or more characteristics characteristic of such risk. In some embodiments, the subject is a patient. In some embodiments, the subject is an individual to whom a diagnostic and / or therapeutic approach is administered and / or has been administered.

[0369] Suffering from: An individual who is "suffering from" a disease, disorder, and / or condition (eg, HIV, HIV-related disorders, etc.) has been diagnosed with and / or exhibits one or more symptoms of the disease, disorder, and / or condition.

[0370] Susceptible to: An individual who is "susceptible" to a disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.) is an individual who is at a higher risk of developing the disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.) than members of the general public. In some embodiments, an individual who is susceptible to a disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.) may not have been diagnosed with the disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.). In some embodiments, an individual who is susceptible to a disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.) may have exhibited symptoms of the disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.). In some embodiments, an individual who is susceptible to a disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.) may not have exhibited symptoms of the disease, condition, and / or disorder (e.g., HIV, HIV-related disorders, etc.). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., HIV, HIV-related disorders, etc.) will develop the disease, disorder, and / or condition (e.g., HIV, HIV-related disorders, etc.). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., HIV, HIV-related disorders, etc.) will not develop the disease, disorder, and / or condition (e.g., HIV, HIV-related disorders, etc.).

[0371] Therapy: The term "therapy" refers to the administration or delivery of an agent or intervention that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect (e.g., one that has been shown to be statistically likely to have such an effect when administered to a relevant population). In some embodiments, a therapeutic agent or therapy is any substance that can be used to alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or features of a disease, disorder, and / or condition (e.g., HIV, HIV-related disorders, etc.). In some embodiments, a therapeutic agent or therapy is a medical intervention (e.g., surgery, radiation, phototherapy) that can be performed to alleviate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or features of a disease, disorder, and / or condition.

[0372] Treatment: As used herein, the term "treat / treatment / treating" refers to any method for partially or completely alleviating, improving, alleviating, suppressing, preventing one or more symptoms or features of a disease, disorder and / or illness (e.g., HIV, HIV-related disorders, etc.), delaying its onset, reducing its severity and / or reducing its incidence. Treatment can be administered to a subject who does not show signs of a disease, disorder and / or illness (e.g., HIV, HIV-related disorders, etc.). In some embodiments, treatment can be administered to a subject who only shows early signs of a disease, disorder and / or illness (e.g., HIV, HIV-related disorders, etc.), for example, for the purpose of reducing the risk of developing the pathology associated with the disease, disorder and / or illness. In some embodiments, treatment can be administered to a subject in the late stages of a disease, disorder and / or illness (e.g., HIV, HIV-related disorders, etc.). DETAILED DESCRIPTION

[0373] I. Human immunodeficiency virus (HIV)

[0374] Human immunodeficiency virus (HIV) is a lentivirus in the retroviridae family. Mature HIV particles are typically round in shape and approximately 100 nm in diameter. They consist of a core (from innermost to outermost) containing two identical single-stranded RNA molecules, a capsid, and an envelope ( Figure 1B (Musumeci et al., Molecules 20.9 (2015): 17511-17532, which is incorporated herein by reference). The envelope is composed of a lipid bilayer and Env proteins. These Env proteins exist as trimers of gp120 surface proteins, anchored to the envelope by gp41 transmembrane proteins. The viral capsid is surrounded by the envelope and comprises a symmetrical outer capsid membrane composed of the matrix protein p17. Within the outer capsid membrane is a conical capsid containing the inner capsid protein p24. The inner capsid is attached to the outer capsid membrane at its conical cone. The inner capsid contains the viral RNA (two identical copies) and the viral enzymes: reverse transcriptase, integrase, and protease. Viruses also contain oligopeptides generated by proteolytic processing of Gag and Gag / Pol precursor proteins p55 and p160 during virion maturation (GAC, Transfusion Medicine Hemotherapy, 43:203-222, 2016, which is incorporated herein by reference).

[0375] There are two main types of HIV: HIV-1 and HIV-2. HIV-1 is the most common type of HIV and accounts for 95% of all infections worldwide. HIV-2 is less common and less contagious. HIV-2 is primarily concentrated in West Africa and surrounding countries.

[0376] HIV-1 and HIV-2 share many similarities, including their intracellular replication pathways, modes of transmission, and the clinical effects that lead to acquired immunodeficiency syndrome (AIDS). However, due to its lower transmissibility, HIV-2 is less likely to progress to AIDS. Consequently, individuals infected with HIV-2 typically experience prolonged periods of no observable disease progression, whereas those infected with HIV-1 progress more rapidly and are more susceptible to AIDS.

[0377] However, once progression begins, the pathological course of both viruses is broadly similar. One difference is that HIV-2 has been found to progress at higher CD4 counts. Furthermore, HIV-2 infection is characterized by lower viral loads, exceeding 10,000 copies / mL, compared to the millions of copies / mL seen with HIV-1. In the case of HIV-2 infection, the subject's immune response tends to be more protective, thus slowing disease progression.

[0378] HIV-1 and HIV-2 are further divided into groups and subtypes. HIV-1 is divided into the major or M group, the unusual or O group, and the non-M / O or N group. The most common group is group M, which is primarily responsible for the global HIV epidemic. The other groups are relatively rare and are found in selected geographic settings, including Gabon, Cameroon, and Equatorial Guinea.

[0379] Group M is further genetically divided into distinct subtypes: A, B, C, D, F, G, H, J, and K. Some of these subtypes combine to form hybrid viruses known as "circulating recombinant forms." Globally, subtype B accounts for 12% of HIV infections. Subtype B is the predominant HIV-1 subtype found in the Americas, Australia, and Western Europe. Consequently, most HIV clinical research to date has focused on these subtypes.

[0380] Despite accounting for nearly 50% of all HIV infections, subtype C has been poorly studied. Subtype C is common in southern African countries, where HIV prevalence is high. Cameroon and the Democratic Republic of the Congo, the regions of origin of HIV-1, are known for their high subtype diversity. However, global subtype distribution patterns are currently changing due to population mixing and migration.

[0381] To date, approximately eight HIV-2 subtypes have been identified. The two main HIV-2 subtypes considered prevalent are A and B. HIV-2 group A infections are primarily seen in West Africa, but a few cases have also been reported in Brazil, Europe, the United States, and India. HIV-2 group B infections are only seen in West Africa.

[0382] Because HIV subtypes may have diversity geographically, ideal therapeutic agents can target and neutralize more than one subtype, even more preferably multiple HIV strains. As discussed further below, anti-HIV antibodies that can bind and at least temporarily neutralize HIV virions have been developed. Nevertheless, this type of anti-HIV antibody still has problems, including administering challenges, antibody persistence in vivo, and viral escape. As described herein, polyribonucleotides and compositions of the present disclosure address these challenges.

[0383] A. HIV genome

[0384] HIV contains two identical copies of the single-stranded DNA encoding its genome. When the virus is integrated into the host cell, the viral RNA is reverse transcribed into double-stranded DNA, thereby causing RNA degradation and integration of double-stranded DNA or proviral DNA into the host genome. Both ends of the HIV genome are flanked by LTR (long terminal repeat) regions, including 5' LTRs encoding transcription promoters. The RNA genome is 9749 nucleotides and includes a 5' cap, a 3' poly (A) tail, and several open reading frames (ORFs) (Wain-Hobson et al., Cell 40 (1): 9-17, 1985, which is incorporated herein by reference).

[0385] The HIV genome includes the following genes: gag, pol, vif, vpr, tat, rev, vpu, env, and nef (see Figure 1A The proteins encoded by gag, pol, and env are viral structural proteins. The proteins encoded by tat and rev are essential regulatory proteins. The proteins encoded by nef, vpr, vif, and vpu are auxiliary regulatory proteins. The gag gene encodes the outer core membrane protein (p17), the capsid protein (p24), the nucleoprotein (p7), the Pr55Gag precursor protein, and the p6 protein. Protein p24 forms the conical capsid, and protein p17 forms the inner membrane layer. Protein p6 is involved in the release of viral particles.

[0386] The pol gene encodes the Pr160GagPol precursor protein, the protease p10, the reverse transcriptase (p51), and RNase H (p15), or both together as the p66 protein, and the integrase p32. Pr160GagPol is the precursor of the viral enzymes p10, p51, and p15. Proteolytic cleavage of Gag (Pr55) and Gag-Pol (Pr160GagPol) produces the protease p10. The protein p51 reverse transcriptase is responsible for transcribing HIV RNA into proviral DNA. When proviral DNA is produced, the protein p55 (RNAse H) is used to degrade the viral RNA in the viral RNA / DNA complex. The protein p32 integrase functions to integrate the proviral DNA into the host cell genome.

[0387] The env gene encodes the precursor protein PrGp160 for two envelope glycoproteins, gp120 (surface protein) and gp41 (transmembrane protein). Proteins gp120 and gp41 are produced by protease cleavage of the precursor protein PrGp160. The gp120 protein plays a role in attaching the virus to the target host cell. The gp41 protein anchors gp120 to the viral membrane and facilitates fusion of the viral and target cell membranes.

[0388] The tat gene encodes the Tat protein p14 (transactivator), which activates transcription of viral genes. The rev gene encodes the Rev protein p19 (RNA splicing regulator), which regulates the export of mRNA (unspliced ​​and partially spliced). The nef gene encodes the Nef protein p27 (negative regulator), which plays a role in HIV replication and enhances the infectivity of the virus in host cells. Protein p27 also serves to downregulate CD4 and HLA on target cells. The vif gene encodes the Vif protein p23 (viral infectivity factor), which functions to produce virus in host cells. The vpr gene encodes the Vpr protein p15 (viral protein r). This protein interacts with the p6 protein and promotes the infectivity of the virus in host cells. The vpu gene encodes the Vpu protein p16 (viral unique protein), which allows for the efficient release of viral particles and controls the degradation of CD4 on target cells. Protein p16 also controls intracellular signaling. The gene vpx encodes the Vpx protein p15 (viral protein x), which interacts with the p6 protein and is important in the early stages of viral replication. The gene tev encodes the Tat / Rev protein p26, which is a fusion protein that regulates the Tat and Rev proteins (GAC, Transfusion Medicine Hemotherapy, 43:203-222, 2016, which is incorporated herein by reference).

[0389] B. Life cycle

[0390] The life cycle of HIV involves the entry of HIV virions into target host cells, reverse transcription of the viral genome, integration into the host genome, and protein maturation. To initiate infection, HIV particles come into contact with target host cells. The surface glycoprotein env gp120 of mature HIV particles binds to the CD4 receptor on the target host cell, thereby initiating additional binding of gp120 to co-receptors (i.e., chemokine receptor 5 (CCR5) or chemokine receptor 4 (CXCR4 of fusion proteins)). The binding of gp120 to CD4 and co-receptors triggers a conformational change in gp120, allowing gp41 to be presented on the viral membrane and to fuse with the plasma membrane of the target host cell. The viral capsid then enters the cytoplasm of the host cell. The capsid is absorbed by the endosome, releasing its contents, i.e., viral RNA. After the virus enters and is released into the target host cell, the virus undergoes reverse transcription, wherein the viral RNA is reverse transcribed into single-stranded cDNA. Next, the RNA strand is degraded by RNase H, and the single-stranded cDNA is converted into double-stranded DNA by the DNA-dependent DNA polymerase activity of reverse transcriptase.

[0391] The double-stranded DNA, or proviral DNA, forms a complex with the integrase enzyme and is transported to the host cell's nucleus, where it randomly inserts itself into the host cell genome. Once integrated into the genome, the proviral genome replicates. The proviral genome can replicate along with the host cell genome as part of cell division, or it can replicate using its own mechanisms. For example, the LTR promoter creates an attachment site for the cellular DNA-dependent RNA polymerase and transcription factors to initiate transcription. The Tat protein accelerates transcription of the proviral DNA.

[0392] The process of entering the target host cell, reverse transcription, integration and protein maturation can be completed in less than 24 hours, and progeny virus particles can be detected within 12 hours after infection. The first progeny virus particles after infection may be released from the infected cells about 24 hours after infection. The infected T cells are usually eliminated by the immune system (for example, by cytotoxic T cells) at a rate of 2-4 days. Because the T cells infected by HIV are destroyed and the production of T cells is restricted, T helper cells will decrease. Proteins nef and tat also inhibit the maturation and replacement of helper T cells. Therefore, over time, HIV infection will lead to immunodeficiency (GAC, Transfusion Medicine Hemotherapy, 43:203-222, 2016, which is incorporated herein by reference).

[0393] C. Transmission and Pathology

[0394] HIV enters the body through intact mucous membranes, damaged skin, or through parenteral inoculation. HIV is most commonly transmitted sexually. HIV can be detected throughout the body approximately 10-14 days after infection, and transmission through the blood or transplanted organs is possible approximately 5-6 days after infection. Clinical symptoms usually appear 3-6 weeks after infection and may include fever, swollen lymph nodes, fatigue, rash, gastrointestinal symptoms, acute neuropathy, myalgia, and / or malaise. However, during this acute phase, some individuals are asymptomatic. These symptoms of acute or primary infection can last 2-6 weeks. This initial symptomatic period is then usually followed by an asymptomatic or sporadic symptomatic phase that may last for several years.

[0395] If untreated, HIV infection leads to progressive CD4+ T cell loss, which can cause a range of immune abnormalities and increase the risk of infectious and neoplastic complications. Furthermore, HIV infection can lead to cardiovascular disease, bone disease, kidney and liver dysfunction, and several other common pathologies.

[0396] Although antiviral therapy (ART) has been developed to treat HIV infection, ART can only prevent new cells from being infected, that is, if the cell already contains viral DNA integrated into its genome, ART cannot eliminate the infection. In addition, HIV establishes a latent infection in CD4+T cells, and the infection can be maintained indefinitely, and some have the ability to self-renew. Once HIV is integrated into the cell genome, it may continue to restart replication in the cell. (Deeks et al. Nature reviews 1.1 2015 and GAC, Transfusion Medicine Hemotherapy, 43: 203-222, 2016, which are incorporated herein by reference).

[0397] D. Treatment strategies

[0398] The development of therapeutic agents targeting HIV faces many challenges. One challenging factor is the heterogeneity of the virus. HIV can be divided into at least two major types (HIV-1, which is found worldwide; and HIV-2, which is mainly found in West Africa), and HIV-1 is further divided into three subgroups (M, N, O and P), and M is further divided into subtype AL. Subtypes can also recombine after co-infection, thereby generating other recombinant subtypes.

[0399] Another challenging factor is the high mutation rate of HIV in vivo. A recent study quantified the spontaneous mutation rate of HIV-1 genome-wide in DNA sequences from peripheral blood mononuclear cells and revealed an extremely high mutation rate of 4.1 ± 1.7 × 10 per base per cell. -3, which is the highest mutation rate ever reported for any biological entity (Cuevas et al., PloS Biol 2015, which is incorporated herein by reference). Therefore, the ability to identify and develop therapeutics that target conserved epitopes across multiple groups and subtypes of HIV sequences, which are constantly mutating, is extremely challenging, and the virus has a unique ability to evade the immune system.

[0400] In addition to its high mutation frequency, HIV presents other challenges to the immune system, making its treatment uniquely challenging. Therapeutic targets on HIV include the HIV envelope protein (HIV Env). However, HIV Env is heavily glycosylated, and the Env site is therefore shielded from therapeutic use by the presence of glycans. Furthermore, Env glycans are derived from the host and can be highly heterogeneous.

[0401] Recent therapeutic strategies involve the use of broadly neutralizing antibodies (bNAbs), which are antibodies that can neutralize a variety of global HIV isolates. Such antibodies have been identified from HIV-infected individuals who are considered "elite neutralizers," which account for <10% of HIV patients (Burton and Hangartner, Ann. Rev. Immunol. 2016, which is incorporated herein by reference). Such antibodies provide insights into potential target epitopes and structures for therapeutic agents. Other advances that contribute to the advancement of therapeutic agents include the generation of stable HIV Env spike trimers (Sanders and Moore, Immunol. Rev. 2017, which is incorporated herein by reference) and the characterization of their structures at high resolution (Ward and Wilson, Immunol. Rev. 2017, which is incorporated herein by reference). Examples of Env sites that are potential targets include the apical site, the high mannose patch in the gp120 region, the gp120-gp41 interface region, the gp41 membrane proximal region (MPER), and the CD4 binding site (see Figure 2 , from McCoy and Burton, Immunol Rev. 275.1 11-20 2017, which is incorporated herein by reference). As therapeutic targets for bNAbs, each of these sites presents unique challenges. For example, bNAbs targeting the gp41-gp120 interface must be able to bind to complex heterogeneous glycans. BNAbs targeting the CD4 binding site of the Env protein have been found to exhibit high levels of somatic hypermutation.

[0402] Nonetheless, among these loci, CD4b is of particular interest because CD4 serves as the primary receptor for viral entry. Certain CD4b bNAbs are characterized by the use of the immunoglobulin heavy chain gene segment IGVH1-2*02, high levels of somatic hypermutation, a five-residue complementarity-determining region 3 (LCDR3) of the light chain, and mimicking of the Env-CD4 interaction. Other CD4b bNAbs are characterized by the use of the immunoglobulin heavy chain gene segment IGVH1-46 (e.g., IGVH1-46*01) and may have a longer LCDR3.

[0403] In addition, the present disclosure also provides polyribonucleotides encoding antibody agents (e.g., bNAbs) that target a wider group of HIV variants and are therefore capable of treating more HIV patients. In addition, the present disclosure also provides compositions for delivering polyribonucleotides encoding antibody agents (e.g., bNAbs) that target various HIV sequences.

[0404] 1. Antiviral treatment for HIV

[0405] At present, HIV infection is mainly treated with antiretroviral therapy (ART). ART is a type of drug that can reduce HIV proliferation, increase CD4 cell count and reduce the risk of transmission to infected individuals. The World Health Organization (WHO) recommends starting ART for all adults infected with HIV, regardless of clinical stage or CD4 cell count (Consolidated guidelines on HIV prevention, testing, treatment, service delivery and monitoring: recommendations for a public health approach. Geneva: World Health Organization; 2021, which is incorporated herein by reference). However, ART is not a curative therapy. If the infected individual stops taking ART, viremia (such as viral load) will rebound rapidly. The high mutation rate of HIV also constrains patients to strictly adhere to their therapy to avoid escape mutants and treatment failure. Therefore, ART is intended to be taken daily by infected subjects for life.

[0406] There are several types of FDA-approved ART treatments for HIV, which work through different mechanisms. Effective management of HIV infection typically involves a combination of at least three ARTs to treat the complex pathogenicity of the disease. The most effective ART combination typically varies depending on the infected individual (see, for example, Bhatti et al., Cureus 2016, which is incorporated herein by reference). Cihlar et al., Current opinion in virology, 2016 (incorporated herein by reference in its entirety) reviewed the ART drug classes used to treat HIV.

[0407] Table 1: Exemplary classes of ART for the treatment of HIV, their mechanisms of action, and exemplary compounds within each class

[0408]

[0409]

[0410] 2. HIV antibody agent

[0411] In addition to ART, anti-HIV antibodies have also been developed. Using anti-HIV antibodies to treat HIV generally requires antibodies to have specific characteristics, including safety, favorable pharmacokinetic profiles, efficient neutralizing activity, and broad neutralizing activity to effectively target the diversity present in HIV virions. As with other HIV therapeutics (including, for example, ART), viral escape of anti-HIV antibodies is a major challenge.

[0412] For example, Barouch et al. infected rhesus macaques with SHIV-SF162P3 (Barouch et al., Nature 503:7475224-228, 2013), which is incorporated herein by reference in its entirety. Rhesus macaques were then treated with three monoclonal antibodies (mAbs): the N332 glycan-dependent mAb PGT121 and the CD4 binding site-specific mAbs 3BNC117 and b12. The mAbs were administered as a mixture at 10 mg / kg each on days 0 and 7; as a mixture alone at 10 mg / kg each on day 0; or as a combination of PGT121 and 3BNC117 alone at 10 mg / kg each. Transient viral suppression was not observed until bNAb levels fell below 10 μg / mL. mAbs were also administered alone to rhesus macaques, and PGT121 alone induced rapid virological control, with most animals rebounding after 6-8 weeks. Rhesus monkeys that received the combination of PGT121 and 3BNC117 received a second dose on day 105 after viral levels rebounded. Viral resuppression was observed, although control was not as durable as with the previous administration.

[0413] Shingai et al. described rhesus macaques infected with SHIV AD8EO (Shingai et al., Nature 503:7475 277-280, 2013, which is incorporated herein by reference in its entirety). Rhesus macaques were then treated with 10-1074 and 3BNC117 mAbs, either alone or in combination. When administered alone at 10 mg / kg 12 weeks after vaccination, both antibodies caused rapid viral suppression, but viral levels rebounded rapidly. Combining the two antibodies in chronically infected animals resulted in a longer period of suppression and increased CD4+ T cell levels, but viral levels subsequently rebounded. In other studies, the two antibodies were pretreated in rhesus macaques alone and found to prevent viral infection. Single genome analysis of the virus that rebounded in 10-074-treated rhesus macaques revealed that a mutation eliminated the gp120 N332 glycan, conferring resistance to the mAb. However, SGA analysis of rebound virus in rhesus macaques treated with 10-074 and 3BNC117 revealed that not all macaques harbored virus that had undergone changes conferring mAb resistance.

[0414] Caskey et al. describe the first human dose escalation phase 1 clinical trial of 3BNC117 (CD4 binding site antibody) (Caskey et al., Nature 522.7557:487-491, 2015, which is incorporated herein by reference in its entirety). Both uninfected and HIV-1 infected individuals participated in the trial. It has been found that 3BNC117 at doses of 1, 3, 10 or 30 mg / kg is generally safe and well tolerated; no grade 3, 4 or serious adverse events were observed. It was observed that the antibody clearance rate in HIV-1 infected individuals was faster than that in uninfected control subjects. The effect of treatment on viral load was dose-dependent; 10 and 30 mg / kg doses reduced viral load by up to 2.5 log. It was observed that regardless of the mAb dose, some individuals developed viral resistance, but others did not. The virus was cloned and sequenced, and G459D was a mutation commonly observed in the 10 mg / kg group, with other mutations showing a longer V5 loop (other mutations described). Both mutations can alter sensitivity to anti-CD4b.

[0415] Caskey et al. also evaluated 10-074, a highly effective mAb that targets the V3 loop of the HIV-1 envelope spike (Caskey et al., Nature Medicine 23.2:185-191, 2017), which is incorporated herein by reference in its entirety. An open-label, Phase 1, first-in-human clinical trial was conducted with 14 uninfected individuals and 19 HIV-1 infected individuals. A single intravenous infusion was administered at 3, 10, or 30 mg / kg. The mAb was found to be generally safe and well tolerated; no grade 3, 4, or serious adverse events were observed. Antibody clearance was observed to be faster in HIV-1 infected individuals than in uninfected controls. Treatment suppressed viral load in individuals with 10-074 susceptible strains, followed by rebound. Single genome sequencing (SGS) of the rebound viruses revealed that all patients who responded to treatment displayed PNGS at position N332 and an intact 324 G(D / N)IR 327 Four weeks after infusion, 91% of the envelope sequences contained amino acid mutations, 97% of which eliminated the PNGS at position 332 by mutations to N332 or S334. 3% of the mutant sequences were at 324 G(D / N)IR 327 The D / N position 325 in the motif showed a change. Most of the mutations at the nucleic acid level were transitions, consistent with reverse transcriptase errors. Neutralization assays showed that the HIV-1 mutants resistant to 10-074 were not resistant to 3BNC117, VRC01, or PGDM1400 (mAbs targeting other regions of HIV-1). Sequencing and scintigraphy performed one week after infusion revealed that the resistant variants were either pre-existing or rapidly emerged.

[0416] Bar et al. conducted two open-label laboratory trials to investigate the safety, side effect profile, pharmacokinetic properties, and antiviral activity of VRC01, a bNAb targeting the CD4 binding site of HIV, in patients who were interrupting antiretroviral therapy (ART) (Bar et al., New England Journal of Medicine 375.21:2037-2050, 2016, which is incorporated herein by reference in its entirety). In one trial, 40 mg / kg was infused three times over a 6-week period; in the other trial, 40 mg / kg was infused eight times over a 6-week period. Treatment was well tolerated, with no grade 3 or higher adverse events observed. Neither trial produced a durable suppression of plasma viremia, although a slight increase in rebound time relative to historical controls was found. Regardless of rebound time, in one trial, participants developed an almost universal increase in resistance to VRC01. Viral isolates showed greater resistance to VRC01 neutralization in samples before treatment than after treatment. VRC01 treatment did not affect sensitivity to neutralization by other bNAbs.

[0417] Mendoza et al. conducted a Phase 1b clinical trial evaluating a combination of 3BNC117 and 10-1074, which were infused at a dose of 30 mg / kg at weeks 0, 3, and 6 (Mendoza et al., Nature 561.7724:479-484, 2018, which is incorporated herein by reference in its entirety). These two bNAbs target independent sites on the HIV-1 envelope spike. Infusions were generally found to be safe and well tolerated, with no serious adverse events reported. The median rebound time after combined bNAb treatment was significantly prolonged. It has been found that the two earliest rebound individuals had previously carried strains resistant to one or another bNAb. The rebound viruses clustered within low-diversity lineages, consistent with the amplification (escape) of 1-2 recurrent viruses. As compared with the 3BNC117 mutation, most rebound viruses were found to contain 10-1074 mutations. However, combined bNAb therapy has been shown to be more effective than single bNAb treatment in curbing viral escape.

[0418] Gautam et al. infected SHIV AD8EORhesus macaques treated with 3BNC117-LS and 10-074-LS mAbs were analyzed (Gautam, Rajeev et al., Nature Medicine 24.5:610-616, 2018, which is incorporated herein by reference in its entirety). M428L and N343S (collectively referred to as LS) are mutations in the mAb fragment domain that increase half-life. The LS mutation had no effect on virus neutralization in in vitro analysis. The LS mAb was administered alone at 20 mg / kg and was well tolerated by all monkeys. The 10-1074-LS receptor showed increased protection against viral infection compared to the 3BNC117-LS receptor, but the LS mutations in both antibodies were more effective than WT. The decay rate of 10-1074-LS in serum was slower than that of 3BNC117-LS. It has been determined that mAb concentration / neutralizing activity can predict the probability of infection. Experiments were performed with only antibody pretreatment followed by viral challenge.

[0419] Schommers et al. characterized an anti-HIV antibody (termed "1-18") in in vitro assays and in HIV-1 infected humanized mice. 1-18 was reported to bind to the CD4 binding site of HIV and to have potent and broad activity against HIV strains. Schommers reported that 1-18 has certain features previously found in other anti-HIV antibodies that appear to contribute to its potency and breadth: (1) 1-18 has an aromatic residue that mimics residue Phe43 of CD4 to target the "Phe43 gp120 pocket," a feature previously reported for the anti-HIV antibody N6; (2) 1-18 contacts the adjacent gp120 protomer, as previously observed in the anti-HIV antibody 3BNC117, but with an increased buried surface area (by inserting six of its residues into CDRH1); and (3) the buried surface area on gp120 is larger than that of other anti-HIV antibodies. Furthermore, 1-18 has been reported to contact conserved residues on HIV gp120 that are not contacted by other anti-HIV antibodies. Schommers hypothesized that these contacts might make 1-18 less reliant on classical CD4 binding site contacts, making viral escape more difficult. Despite this, Schommers observed that a small number of HIV strains were resistant to 1-18.

[0420] Taken together, the above data suggest that administration of antibodies can be effective in treating or preventing HIV. However, the difficulty of targeting such mutable viruses is evident from the above studies as well as studies showing that: (1) when a single broadly neutralizing antibody (bNAb) is used for therapy, HIV develops resistance to the therapy within weeks (Bar et al., Effect of HIV Antibody VRC01 on Viral Rebound after Treatment Interruption, N. Engl. J. Med. 375, 2037-2050 (2016); Caskey et al., Viraemia suppressed in HIV-1-infected humans by broadly neutralizing antibody 3BNC117. Nature 522, 487-491 (2015); Caskey et al., Antibody 10-1074 suppresses viremia in HIV-1-infected individuals. Nat. Med. 23, 185-191 (2017); Klein et al., HIV therapy by a combination of broadly neutralizing antibodies in humanized mice, Nature 492, 118-122 (2012); Lynch et al., Virologic effects of broadly neutralizing antibody VRC01 administration during chronic HIV-1 infection, Sci. Transl. Med. 7, 319ra206 (2015); Scheid et al., HIV-1 antibody 3BNC117 suppresses viral rebound in humans during treatment interruption, Nature 535, 556-560 (2016), each of which is incorporated herein by reference in its entirety), and (2) certain antibody combinations improve viral control by preventing the early development of resistance (Bar-On et al., Safety and antiviral activity of combination HIV-1 broadly neutralizing antibodies in viremic individuals, Nat. Med.24, 1701-1707 (2018); Klein et al., 2012; Mendoza et al., Combination therapy with anti-HIV-1 antibodies maintains viral suppression, Nature 561, 479-484 (2018), each of which is incorporated herein by reference in its entirety). Viral rebound was observed with some of these antibodies, suggesting that they may be effective only for a limited time, for example, before HIV escape mutations occur.

[0421] Therefore, it is still necessary to avoid viral escape and keep HIV neutralization effective treatment and preventive therapeutic agents.As discussed herein, the present disclosure provides the technology that can be used for administering the polyribonucleotide of encoding one or more antibody agents (for example, anti-HIV antibody agents) to the subject.Use technology and method as described herein to allow, for example, to produce different antibody agents from polyribonucleotide simultaneously.The form of antibody agent has been designed to minimize or eliminate the risk of immunoglobulin chain mispairing.Can combine multiple antibody agent formula as described herein (for example, including 1-18 antibody agent) and allow to develop a kind of composition (for example, pharmaceutical composition), described composition delivers multiple antibody agents together so that they can be in conjunction with the different epitopes of HIV virus, thereby minimize the virus escape caused by mutation and increase overall efficacy.

[0422] II. Polyribonucleotides for delivery of antibody agents

[0423] In addition, the present disclosure utilizes RNA technology as a way to express antibody agents directly in a subject, which is a novel class of antibody-based therapeutic methods. In some embodiments, the polyribonucleotides as described herein encode immunoglobulin chains of the antibody agent.

[0424] In some embodiments, the antibody agent targets HIV. In some embodiments, the antibody agent targeting HIV specifically binds to a particular epitope of an HIV polypeptide. For example, in some embodiments, the antibody agent specifically binds to an epitope that includes the CD4 binding site or a portion thereof. See Figure 2 , from McCoy and Burton, Immunol Rev. 275.1 11-20, 2017, which is incorporated herein by reference.

[0425] In some embodiments, the antibody agent may have at least about 10 -4 M, at least about 10 -5 M, at least about 10 -6 M, at least about 10 -7 M, at least about 10 -8 M, at least about 10-9 In some embodiments, the HIV antibody agent selectively binds to the target epitope of HIV, such that the binding between the HIV antibody agent and the target epitope is greater than 2 times, greater than 5 times, greater than 10 times, or greater than 100 times compared to the binding of the HIV antibody agent to a non-target epitope. In some embodiments, the HIV antibody agent can have binding affinity for HIV epitopes and variants of the HIV epitope. It will be understood by those skilled in the art that in some cases, binding affinity (e.g., as measured by the dissociation constant) may be affected by non-covalent intermolecular interactions, such as hydrogen bonding, electrostatic interactions, hydrophobic forces, and van der Waals forces between the two molecules. Alternatively or in addition, the binding affinity between the ligand and its target molecule may be affected by the presence of other molecules. Those skilled in the art will be familiar with various techniques for measuring binding affinity and / or dissociation constants in accordance with the present disclosure, including, for example, but not limited to, ELISA, gel shift assays, pull-down assays, equilibrium dialysis, analytical ultracentrifugation, surface plasmon resonance (SPR), bio-layer interferometry, grating coupling interferometry, and spectroscopic analysis.

[0426] In some embodiments, the antibody agent targeting HIV may comprise or be derived from a broadly neutralizing antibody (bNAb). In some embodiments, the antibody agent targeting HIV can be any one of the HIV targeting antibodies described in Barouch et al., Nature 503:7475224-228, 2013, Shingai et al., Nature 503:7475 277-280, 2013, Caskey et al., Nature 522.7557:487-491, 2015, Caskey et al., Nature Medicine 23.2:185-191, 2017, Bar et al., New England Journal of Medicine 375.21:2037-2050, 2016, Mendoza et al., Nature 561.7724:479-484, 2018, Gautam, Rajeev et al., Nature Medicine 24.5:610-616, 2018, the contents of each of which are incorporated herein by reference in their entirety for the purposes described herein.

[0427] In some embodiments, the antibody agent targeting HIV can be, for example, 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, PGDM1400, a fragment thereof, or a combination thereof. Exemplary anti-HIV antibodies that can be used in the compositions described herein include, but are not limited to, 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, PGDM1400, a fragment thereof, or a combination thereof. For example, in some embodiments, polyribonucleotides as described herein can comprise one or more heavy chain complementary determining regions (HCDRs) (e.g., HCDR1, HCDR2, and / or HCDR3) from 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400. In some embodiments, polyribonucleotides as described herein can comprise HCDR1, HCDR2, and HCDR3 from 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400. In some embodiments, polyribonucleotides as described herein can comprise a heavy chain variable domain from 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400. In some embodiments, polyribonucleotides as described herein can comprise one or more light chain complementary determining regions (LCDRs) (e.g., LCDR1, LCDR2, and / or LCDR3) from 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400. In some embodiments, the polyribonucleotides as described herein can comprise LCDR1, LCDR2, and LCDR3 from 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400. In some embodiments, the polyribonucleotides as described herein can comprise a light chain variable domain from 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400.

[0428] In some embodiments, a plurality of polyribonucleotides each encoding an immunoglobulin chain of an antibody agent can be used to deliver (e.g., by administering to a subject) two or more antibody agents (e.g., 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400, or a fragment or variant thereof). In some embodiments, a plurality of polyribonucleotides each encoding an immunoglobulin chain of an antibody agent can be used to deliver (e.g., by administering to a subject) three or more antibody agents (e.g., 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400, or a fragment or variant thereof). In some embodiments, a plurality of polyribonucleotides each encoding an immunoglobulin chain of an antibody agent can be used to deliver (e.g., by administering to a subject) four or more antibody agents (e.g., 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400, or a fragment or variant thereof). In some embodiments, a plurality of polyribonucleotides each encoding an immunoglobulin chain of an antibody agent can be used to deliver (e.g., by administering to a subject) two, three, four, five, or six antibody agents (e.g., 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400, or a fragment or variant thereof).

[0429] In some embodiments, the antibody agent encoded by one or more polyribonucleotides described herein includes all or part of the 1-18 antibody. In some embodiments, the antibody agent encoded by one or more polyribonucleotides provided herein includes all or part of the 1-18 antibody. In some embodiments, the antibody agent comprises a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof. In some embodiments, the antibody agent comprises a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12). In some embodiments, the antibody agent comprises a light chain variable domain comprising: (i) LCDR1 (QGL DSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, the antibody agent comprises a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21). In some embodiments, the antibody agent comprises (a) a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO:6); (ii) HCDR2 (ISPHFARP; SEQ ID NO:9); (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO:12); or (iv) a combination thereof; and (b) a light chain variable domain comprising: (i) LCDR1 (QGL DSSH; SEQ ID NO:15); (ii) LCDR2 (GTS; SEQ ID NO:18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO:21); or (iv) a combination thereof.In some embodiments, the antibody agent comprises (a) a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO:6); (ii) HCDR2 (ISPHFARP; SEQ ID NO:9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO:12); and (b) a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO:15); (ii) LCDR2 (GTS; SEQ ID NO:18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO:21).

[0430] In some embodiments, the polyribonucleotides described herein encode all or part of 1-18 antibodies. In some embodiments, the polyribonucleotides described herein encode immunoglobulin chains comprising heavy chain variable domains, wherein the heavy chain variable domains comprise: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNYH MDV; SEQ ID NO: 12); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode immunoglobulin chains comprising heavy chain variable domains, wherein the heavy chain variable domains comprise: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12). In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21). In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DD PYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof; and the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof.In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPF GDRAPHYNYHMDV; SEQ ID NO: 12); and the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GT S; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21). In some embodiments, the polyribonucleotides described herein encode two immunoglobulin chains: a first immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFG DRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof; and a second immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode two immunoglobulin chains: a first immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HC DR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); and a second immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21).

[0431] In some embodiments, the antibody agent encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 24. In some embodiments, the antibody agent encoded by one or more polyribonucleotides provided herein comprises a light chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 29. In some embodiments, the antibody agent comprises a heavy chain variable domain represented by SEQ ID NO: 24. In some embodiments, the antibody agent comprises a light chain variable domain represented by SEQ ID NO: 29.

[0432] In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 24. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 29. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 24, and wherein the light chain variable domain has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 29.

[0433] In some embodiments, the polyribonucleotide encoding antibody agent as described herein is an immunoglobulin chain, wherein the immunoglobulin chain comprises a heavy chain variable (VH) domain. In some embodiments, the VH domain comprises the VH domain of an 1-18 antibody. In some embodiments, the polyribonucleotide encoding antibody agent is selected from the VH domain of a polyribonucleotide encoding antibody: PGT121, 3BNC117, b12, 10-1074, 10-1074-LS, 10E8, VRC01, VRC07-523, or PGDM1400 (e.g., as described herein).

[0434] In some embodiments, the polyribonucleotide comprises a VH domain encoding sequence comprising (a) a HCDR1 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 7; (b) a HCDR2 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 10; (c) a HCDR3 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 13, or (d) a combination thereof. In some embodiments, the polyribonucleotide comprises a VH domain encoding sequence comprising (a) a HCDR1 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 7; (b) a HCDR2 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 10; and (c) a HCDR3 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 13. In some embodiments, the polyribonucleotide encodes a VH domain and comprises a VH coding sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 25 or 27. In some embodiments, the polyribonucleotide encodes a VH domain and comprises a VH coding sequence according to SEQ ID NO: 25 or 27.

[0435] In some embodiments, polyribonucleotides as described herein include the immunoglobulin chains of antibody agents, wherein the immunoglobulin chains include light chain variable (VL) domains. In some embodiments, the VL domains include the VL domains of 1-18 antibodies. In some embodiments, polyribonucleotide encoding is selected from the VL domains of following antibodies: PGT121, 3BNC117, b12, 10-1074, 10-1074-LS, 10E8, VRC01, VRC07-523 or PGDM1400 (for example, as described herein).

[0436] In some embodiments, the polyribonucleotide comprises one or more coding regions for an immunoglobulin chain encoding an antibody agent, wherein the immunoglobulin chain comprises a light chain variable (VL) domain. In some embodiments, the polyribonucleotide comprises a VL domain coding sequence comprising (a) a LCDR1 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 16; (b) a LCDR2 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 19 (GGCACCAGC); (c) a LCDR3 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 22; or (d) a combination thereof. In some embodiments, the polyribonucleotide comprises a VL domain encoding sequence comprising (a) a LCDR1 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 16; (b) a LCDR2 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 19 (GGCACCAGC); and (c) a LCDR3 encoding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 22. In some embodiments, the polyribonucleotide encodes a VL domain and comprises a VL encoding sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 30. In some embodiments, the polyribonucleotide encodes a VL domain and comprises a VL encoding sequence according to SEQ ID NO: 30.

[0437] In some embodiments, the antibody agent is formed from one, two, three, or four immunoglobulin chains.

[0438] In some embodiments, the polyribonucleotides as described herein encode a single immunoglobulin chain. In some embodiments, the first polyribonucleotide encodes the first immunoglobulin chain of an antibody agent. In some embodiments, the first polyribonucleotide encodes the first immunoglobulin chain of an antibody agent and the second polyribonucleotide encodes the second immunoglobulin chain of an antibody agent. In some embodiments, the first polyribonucleotide encodes the first immunoglobulin chain of an antibody agent, the second polyribonucleotide encodes the second immunoglobulin chain of an antibody agent, and the third polyribonucleotide encodes the third immunoglobulin chain of an antibody agent. In some embodiments, the first polyribonucleotide encodes the first immunoglobulin chain of an antibody agent, the second polyribonucleotide encodes the second immunoglobulin chain of an antibody agent, the third polyribonucleotide encodes the third immunoglobulin chain of an antibody agent, and the fourth polyribonucleotide encodes the fourth immunoglobulin chain of an antibody agent.

[0439] In some embodiments, polyribonucleotide encoding two immunoglobulin chains as described herein.In some embodiments, single polyribonucleotide can include the first coding region of the first immunoglobulin chain of encoding antibody and the second coding region of the second immunoglobulin chain of encoding antibody.In some embodiments, the first coding region and the second coding region are separated by internal ribosome entry side (IRES), internal promoter or peptide sequence (such as " from cleavage " 2A or 2A sample sequence) (see, for example, Szymczak et al. Nat Biotechnol 22:589, May, 2004; ePub April 4, 2004, it is incorporated herein by reference) to produce the first immunoglobulin chain and the second immunoglobulin chain from single polyribonucleotide.

[0440] The antibody agent encoded by one or more polyribonucleotides described herein can be in various forms as described herein. Exemplary types of antibody agents include, but are not limited to, monoclonal antibodies or polyclonal antibodies. In some embodiments, the antibody agent may include one or more humanized, chimeric, etc. sequence elements, as known in the art. In some embodiments, the antibody agent used in accordance with the present disclosure is selected from, but not limited to, the following forms: complete IgG, IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies (e.g., etc.); CrossMab (e.g., CrossMab CH1-CLx ;CrossMab CH1-CLcv ; Bispecific CrossMab with a knob-and-hole CH1-CLx ); antibody fragments, such as Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments and isolated complementarity determining regions (CDRs) or collections thereof; single-chain Fv (scFv); scFv-Fc fusions; polypeptide-Fc fusions; single domain antibodies (e.g., shark single domain antibodies, such as IgNAR or fragments thereof); camelid antibodies; masked antibodies (e.g., ); small modular immunopharmaceuticals ("SMIPsTM"); single-chain or tandem bifunctional antibodies VHH; small antibodies; Ankyrin repeat protein or DART;TCR-like antibody; MicroProteins; as well as In some embodiments, immunoglobulin chains and / or fragments of such antibodies may be used in combination, for example, a scFv-Fc arm in combination with a conventional antibody arm.

[0441] Exemplary forms that can be used in accordance with the present disclosure are described further below.

[0442] A. Conventional antibodies

[0443] In some embodiments, polyribonucleotides as herein described can be used for expressing conventional antibodies. As used herein, "conventional antibodies" refer to antibody agents comprising two heavy chains and two light chains (see, e.g., Figure 5A and Figure 4A ). Each heavy chain includes a heavy chain variable domain operably linked to one or more heavy chain constant domains. In some embodiments, the one or more heavy chain constant domains include a CH1 domain, a hinge domain, a CH2 domain, a CH3 domain, or a combination thereof. In some cases, the one or more heavy chain constant domains include a CH1 domain, a hinge domain, a CH2 domain, a CH3 domain, a CH4 domain, or a combination thereof. Each light chain includes a light chain variable domain operably linked to a light chain constant domain.

[0444] Typically, the heavy chain variable domain and the light chain variable domain can be further subdivided into variable regions, referred to as complementary determining regions (CDRs), interspersed with more conservative regions, referred to as framework regions (FRs). Such heavy chain variable domains and light chain variable domains can each include, for example, three CDRs and four framework regions, arranged in the following order from amino-terminal to carboxyl-terminal: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4, one or more of which can be engineered as described herein. The CDRs in the heavy chain are designated as "HCDR1," "HCDR2," and "HCDR3," respectively, and the CDRs in the light chain are designated as "LCDR1," "LCDR2," and "LCDR3," respectively.

[0445] Conventional antibodies as described herein can comprise any of the five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM. In some embodiments, conventional antibodies comprise IgG or IgA antibodies. In some embodiments, conventional antibodies as described herein comprise a specific isotype selected from the IgA and IgG isotype groups: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. In addition, in some embodiments, conventional antibodies can include any specific heavy chain constant domains corresponding to different classes of immunoglobulins, including α, δ, ε, γ, and μ, respectively. In some embodiments, conventional antibodies are complete IgG1 antibodies or other antibody classes or isotypes as described herein. (See, e.g., Hudson et al., Nat. Med., 9:129-134 (2003); Pluckthun, The Pharmacology of Monoclonal Antibodies, Vol. 113, pp. 269-315 (1994); Hollinger et al., Proc. Natl. Acad. Sci. USA, 90:6444-6448 (1993); WO 93 / 01161; and U.S. Pat. Nos. 5,571,894, 5,869,046, 6,248,516, and 5,587,458, each of which is incorporated herein by reference.) In addition to the various isotypes, allelic variation exists between IgG subclasses, giving rise to allotypic variants or allotypes. The IgG antibody agents described herein may comprise a specific allotype, including but not limited to G1m3, G1m17, G1m17,1 or G1m17,1,2 or G1m3,1 (see Vidarsson et al., Front. Immunol, 5(520):1-17, 2014, which is incorporated herein by reference in its entirety).

[0446] The Fc region of a conventional antibody binds to elements of the complement system and also to receptors on effector cells (including, for example, effector cells that mediate cytotoxicity). As is known in the art, the affinity and / or other binding properties of the Fc region for Fc receptors can be modulated by glycosylation or other modifications. In some embodiments, conventional antibodies produced and / or utilized according to the present invention include glycosylated Fc domains, including Fc domains with such glycosylation that are modified or engineered. In some embodiments, conventional antibodies are naturally produced (e.g., produced by an organism in reaction with an antigen), or produced by recombinant engineering, chemical synthesis, or other artificial systems or methods. In some embodiments, conventional antibodies are polyclonal; in some embodiments, conventional antibodies are monoclonal. In some embodiments, conventional antibodies have constant region sequences that are unique to mouse, rabbit, primate, or human antibodies. In some embodiments, conventional antibody sequence elements are humanized, primatized, chimeric, etc., as known in the art.

[0447] Conventional antibodies as described herein are antibodies having a structure generally similar to that of a native antibody or having heavy chains containing an Fc region as defined herein.

[0448] In some embodiments, conventional antibodies encoded by one or more polyribonucleotides provided herein include all or part of 1-18 antibodies. In some embodiments, conventional antibodies comprise a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof. In some embodiments, conventional antibodies comprise a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12). In some embodiments, a conventional antibody comprises a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, a conventional antibody comprises a light chain variable domain comprising: (i) LCDR1 (QGLDS SH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21). In some embodiments, a conventional antibody comprises (a) a heavy chain variable domain comprising: (i) HC DR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHF ARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof; and (b) a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof.In some embodiments, a conventional antibody comprises (a) a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SE QID NO: 12); and (b) a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SE Q ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21).

[0449] In some embodiments, the polyribonucleotides described herein encode the immunoglobulin chains of conventional antibodies, wherein the immunoglobulin chains comprise a heavy chain variable domain, and wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HC DR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode the immunoglobulin chains of conventional antibodies, wherein the immunoglobulin chains comprise a heavy chain variable domain, and wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12). In some embodiments, the polyribonucleotides described herein encode the immunoglobulin chains of conventional antibodies, wherein the immunoglobulin chains comprise a light chain variable domain, and wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR 2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode the immunoglobulin chains of conventional antibodies, wherein the immunoglobulin chains comprise a light chain variable domain, and wherein the light chain variable domain comprises: (i) LCDR1 (QG LDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21).In some embodiments, the polyribonucleotides described herein encode two immunoglobulin chains of a conventional antibody: a first immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNY HMDV; SEQ ID NO: 12); or (iv) a combination thereof; and a second immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode two immunoglobulin chains of a conventional antibody: a first immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HC DR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); and a second immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21).

[0450] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 24. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides provided herein comprises a light chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 29. In some embodiments, a conventional antibody comprises a heavy chain variable domain represented by SEQ ID NO: 24. In some embodiments, a conventional antibody comprises a light chain variable domain represented by SEQ ID NO: 29.

[0451] In some embodiments, the polyribonucleotides described herein encode the immunoglobulin chains of conventional antibodies, wherein the immunoglobulin chains comprise a heavy chain variable domain, and the heavy chain variable domain has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with the amino acid sequence represented by SEQ ID NO: 24. In some embodiments, the polyribonucleotides described herein encode the immunoglobulin chains of conventional antibodies, wherein the immunoglobulin chains comprise a light chain variable domain, and the light chain variable domain has at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity with the amino acid sequence represented by SEQ ID NO: 29.

[0452] Conventional antibodies encoded by one or more polyribonucleotides as described herein may comprise one or more heavy chain constant domains. In some embodiments, one or more heavy chain constant domains comprise a CH3 domain. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH3 domain comprising a G1m3, G1m17, or Glm17,1 allotype. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH3 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identical to the sequence represented by any one of SEQ ID NOs: 68, 71, 74, 77, 80, 83, 86, or 89. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH3 domain having an amino acid sequence represented by any one of SEQ ID NOs: 68, 71, 74, 77, 80, 83, 86, or 89.

[0453] Conventional antibodies encoded by one or more polyribonucleotides as described herein may include one or more heavy chain constant domains, and the heavy chain constant domains include amino acid modifications (e.g., substitutions or deletions) at one or more amino acid positions. For example, conventional antibodies encoded by one or more polyribonucleotides as described herein may include L / S mutations (FcRn combinations for enhanced) in the CH3 region (see Zalevsky J et al. Nat Biotechnol. 2010, which are incorporated herein by reference). Such mutations are labeled as M428L and N434S according to EU numbering, and are referred to herein as "LS" or "L / S" (see e.g., Figure 5C In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an E294 deletion (for Fc hypersialylation) (see Bas M et al. J Immunol 2019, which is incorporated herein by reference).

[0454] The present disclosure also provides techniques that can be used to express antibody agents, e.g., Figure 3As shown or described below: Stadler et al. (2016) Oncoimmunology 5(3):e1091555; and / or Stadler et al. (2017) Nature Medicine 23(7):815-817. There are challenges in producing multiple antibody agents from a single composition (e.g., a composition comprising polyribonucleotides sufficient to encode multiple antibody agents), particularly because random pairing of different antibody heavy chains and light chains can produce unwanted antibody species. Due to the presence of mispaired byproducts and the significant reduction in yield, complex purification procedures are required in these cases to separate the desired antibody agents (see, for example, Morrison, SL, Nature Biotech. 25, 1233-1234, 2007, which is incorporated herein by reference). In general, if recombinant expression technology is used, the problem of mispaired byproducts still exists. One method for solving the problem of mispaired byproducts is called "knob-in-hole technology" (KIH), which aims to modify the contact interface by introducing mutations in the CH3 domain, thereby forcing two different antibody heavy chains to pair. On one chain, bulky amino acids are replaced by amino acids with short side chains to form a "hole", and amino acids with large side chains are introduced into the other CH3 domain to form a "knob". By co-expressing these two heavy chains with two light chains, it was observed that the yield of heterodimer formation was higher than that of homodimers (see Ridgway, JB et al., Protein Eng. 9, 617-621, 1996; and WO 96 / 027011, which are incorporated herein by reference). In some embodiments, the antibody agents described herein utilize KIH technology, as described, for example, in WO 1998 / 050431, which is incorporated herein by reference in its entirety. As described herein, the antibody agent may comprise certain mutations utilizing KIH technology, including but not limited to CH3 modifications. In some embodiments, the antibody agent comprises a CH3 domain comprising one or more of the following mutations: Y349C, T366S, L368A, and Y407V (according to EU numbering). In some embodiments, the antibody agent comprises a CH3 domain, wherein the CH3 domain comprises each of the following mutations: Y349C, T366S, L368A, and Y407V (according to EU numbering). Such a combination of mutations is referred to herein as "cah". In some embodiments, the antibody agent comprises a CH3 domain, wherein the CH3 domain comprises one or more mutations selected from the group consisting of S354C and T366W (according to EU numbering). In some embodiments, the antibody agent comprises a CH3 domain, wherein the CH3 domain comprises each of the following mutations: S354C and T366W (according to EU numbering). Such a combination of CH3 mutations is referred to herein as "cak".

[0455] Thus, in some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH3 domain comprising one or more of the following mutations: Y349C, T366S, L368A, and Y407V (according to EU numbering). In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH3 domain comprising one or more mutations selected from the group consisting of S354C and T366W (according to EU numbering).

[0456] In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one of the following substitution mutations: M428, N434S, or a combination thereof (e.g., an "L / S" mutation). In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence comprising any one of SEQ ID NOs: 75 and 78. In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one or more of the following substitution mutations: Y349C, T366S, L368A, and Y407V (according to EU numbering). In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NOs: 81 and 84. In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one or both of the following substitution mutations: S354C and T366W (according to EU numbering). In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NOs: 87 and 90.

[0457] In some embodiments, the polyribonucleotide encodes an immunoglobulin chain comprising a VH domain operably linked to one or more constant domains, wherein the one or more constant domains comprise a CH3 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NO: 69. In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence encoding a CH3 domain comprising a G1m3, G1m17, or Glm17,1 allotype. In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence comprising any one of SEQ ID NOs: 69 and 72.

[0458] In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH1 domain. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH1 domain comprising a G1m3 allotype. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH1 domain comprising a G1m17 allotype. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH1 domain comprising amino acids represented by SEQ ID NO: 38 or 41.

[0459] In some embodiments, the polyribonucleotide encodes an immunoglobulin chain comprising a VH domain operably linked to one or more constant domains, wherein the one or more constant domains comprise a CH1 domain. In some embodiments, the polyribonucleotide comprises a CH1 ribonucleic acid sequence according to SEQ ID NO: 39. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising a G1m3 allotype. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising a G1m17 allotype. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence according to SEQ ID NO: 39 or 42.

[0460] In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more mutations. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more added serine residues. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising two additional serine residues added (referred to herein as "SS"). In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 45 or 48. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence represented by SEQ ID NO: 45 or 48. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more charge variation mutations. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more substitution mutations selected from the group consisting of K147E, K213D, or a combination thereof. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 51 or 866. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence according to SEQ ID NO: 51 or 866.

[0461] In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein include a hinge domain. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein include a hinge domain, and the hinge domain includes the amino acid sequence represented by SEQ ID NO: 104 (herein referred to as "hinge" in Table 2 and Table 4).

[0462] In some embodiments, polyribonucleotide encoding hinge domain. In some embodiments, polyribonucleotide encoding SEQ ID NO: hinge ribonucleic acid sequence represented by 105. In some embodiments, polyribonucleotide encoding hinge domain, described hinge domain includes amino acid modification, and described amino acid modification includes the disappearance of one or more amino acid residues. In some embodiments, polyribonucleotide encoding hinge domain, described hinge domain includes amino acid modification, and described amino acid modification includes conventional Ig hinge domain (represented by SEQ ID NO: 104) missing amino acid residue EPKSC. Such modification is referred to as " hinge _del " or " ΔEPKSC " in this article. In some embodiments, polyribonucleotide encoding SEQ ID NO: hinge ribonucleic acid sequence represented by 111. In some embodiments, polyribonucleotide encoding hinge domain, described hinge domain includes amino acid modification, and described amino acid modification includes C220S mutation (according to EU numbering). Such mutant hinge domain is referred to as " hinge _S " or " C / S " in this article. In some embodiments, the polyribonucleotide encodes the hinge ribonucleic acid sequence represented by SEQ ID NO:108.

[0463] In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH2 domain. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to that represented by SEQ ID NO: 53. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH2 domain having an amino acid sequence represented by SEQ ID NO: 53.

[0464] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH2 domain having one or more mutations (e.g., relative to SEQ ID NO: 53). For example, in some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises one or more of the following mutations: G236A, A330L, and I332E (according to EU numbering). In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises the following mutations: G236A, A330L, and I332E (according to EU numbering), referred to herein as "GAALIE". Such mutations in the CH2 domain are associated with increased affinity for Fc receptors FcgRIIA and FcgRIII, thereby enhancing antibody effector function.

[0465] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 56. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO: 56.

[0466] In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise one or more mutations selected from the group consisting of G236A and I332E (according to EU numbering). In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise mutations selected from the group consisting of G236A and I332E (according to EU numbering), referred to herein as "GAIE". In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 59. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise a CH2 domain having an amino acid sequence represented by SEQ ID NO: 59.

[0467] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises the mutation: G236A (according to EU numbering), referred to herein as "GA". In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 62. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO: 62.

[0468] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises the mutation: I332E (according to EU numbering), referred to herein as "IE". In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 65. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO: 65.

[0469] In some embodiments, the polyribonucleotide encodes an immunoglobulin chain comprising a VH domain operably linked to one or more constant domains, wherein the one or more constant domains comprise a CH2 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NO: 54. In some embodiments, the CH2 ribonucleic acid encodes a CH2 domain having one or more amino acid substitution mutations. For example, in some embodiments, the CH2 ribonucleic acid sequence encodes one or more of the following mutations: G236A, A330L, and I332E (according to EU numbering), referred to herein as "GAALIE". Such mutations in the CH2 domain are associated with increased affinity for Fc receptors FcgRIIA and FcgRIII, thereby enhancing antibody effector function. In some embodiments, the CH2 ribonucleic acid sequence comprises or consists of a sequence according to SEQ ID NO: 57. In some embodiments, the CH2 ribonucleic acid sequence encodes one or more of the following mutations: G236A and I332E (according to EU numbering), referred to herein as "GAIE". In some embodiments, the CH2 ribonucleic acid sequence comprises a sequence according to SEQ ID NO: 60. In some embodiments, the CH2 ribonucleic acid sequence encodes the mutation G236A (according to EU numbering), referred to herein as "GA". In some embodiments, the CH2 ribonucleic acid sequence comprises a sequence according to SEQ ID NO: 63. In some embodiments, the CH2 ribonucleic acid sequence encodes the mutation I332E (according to EU numbering), referred to herein as "IE". In some embodiments, the CH2 ribonucleic acid sequence comprises a sequence according to SEQ ID NO: 66. In some embodiments, the CH2 ribonucleic acid sequence encodes a CH2 domain comprising a deletion E294 (according to EU numbering).

[0470] In some embodiments, the conventional antibodies encoded by one or more polyribonucleotides as described herein include a signal peptide comprising a human signal peptide. In some embodiments, the conventional antibodies encoded by one or more polyribonucleotides as described herein include a signal peptide comprising a human signal peptide.

[0471] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a light chain constant domain, wherein the light chain constant domain comprises a kappa light chain constant domain. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a kappa light chain constant domain having an amino acid sequence that is at least 80, 85, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 92. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a kappa light chain constant domain having an amino acid sequence represented by SEQ ID NO: 92. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises a lambda chain variable domain.

[0472] In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise immunoglobulin chains (e.g., immunoglobulin heavy chains) encoded by nucleic acid sequences having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to sequences represented by SEQ ID NOs: 113-160. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise immunoglobulin chains (e.g., immunoglobulin heavy chains) encoded by nucleic acid sequences represented by any one of SEQ ID NOs: 113-160. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 449. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) encoded by a nucleic acid sequence represented by SEQ ID NO: 449.

[0473] In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise immunoglobulin chains (e.g., immunoglobulin heavy chains) encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a sequence represented by any one of SEQ ID NOs: 613, 616, 622 and 625. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides as described herein comprise immunoglobulin chains (e.g., immunoglobulin light chains) encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a sequence represented by NO: 619. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin heavy chain) encoded by a nucleic acid sequence represented by any one of SEQ ID NOs: 613, 616, 622, and 625. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) encoded by a nucleic acid sequence represented by NO: 619.

[0474] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin heavy chain) comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to a sequence represented by any one of SEQ ID NOs: 614, 617, 623 and 626. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin heavy chain) comprising an amino acid sequence represented by any one of SEQ ID NOs: 614, 617, 623 and 626. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 620. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides as described herein comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) comprising an amino acid sequence represented by SEQ ID NO: 620.

[0475] Exemplary immunoglobulin chain (eg, immunoglobulin heavy or light chain) configurations of conventional antibodies as described herein are shown in Table 2 below.

[0476] Table 2: Exemplary Immunoglobulin Chain Configurations

[0477]

[0478]

[0479]

[0480] B.CrossMab CH1-CLx

[0481] The present disclosure also provides technologies that can be used to deliver and express antibody agents as described herein in the form of "CrossMab" (see, e.g., WO 2015 / 101588A1, WO 2009 / 080253A1, and Schaefer, W. et al., PNAS, 108, 11187-1191, 2011, which are incorporated herein by reference in their entireties). In some embodiments, an antibody agent in the form of a CrossMab contains a CL-CH1 crossover in one or both binding arms (referred to herein as a "CrossMab"). CH1-CLxSuch modifications reduce the formation of side products due to mispairing of the light chain of a first antibody that specifically binds to a first antigen with the incorrect heavy chain of a second antibody that specifically binds to a second antigen (when compared to methods without such domain swapping).

[0482] In some embodiments, the antibody agent encoded by one or more polyribonucleotides provided herein comprises a first immunoglobulin chain and a second immunoglobulin chain. In some embodiments, the polyribonucleotide can encode a CrossMab as described herein. CH1-CLx The first immunoglobulin chain and the second immunoglobulin chain of the antibody agent. In some embodiments, the polyribonucleotide encoding the first immunoglobulin chain comprises a ribonucleic acid sequence encoding a VH domain, a CL domain, a hinge domain, a CH2 domain, and a CH3 domain. In some embodiments, the polyribonucleotide encoding the second immunoglobulin chain comprises a ribonucleic acid sequence encoding a light chain variable (VL) domain and a CH1 domain (see, e.g., Figure 4B In some embodiments, the CrossMab CH1-CLx The polyribonucleotide of the antibody agent comprises a ribonucleic acid sequence encoding any one of the immunoglobulin chain configurations in Table 3, corresponding to SEQ ID NO: 161-208. In some embodiments, the polyribonucleotide encoding CrossMab CH1-CLx The polyribonucleotide sequence of the antibody comprises a ribonucleic acid sequence encoding any one of the immunoglobulin chain configurations in Table 3, corresponding to SEQ ID NOs: 450 and 451.

[0483] In some embodiments, CrossMab CH1-CLx The antibody agent can be encoded by two separate polyribonucleotides: a first polyribonucleotide comprising a coding region encoding (in 5' to 3' order): a heavy chain variable domain (VH), a light chain constant region (CL), a hinge region, a CH2 domain, and a CH3 domain (see, e.g., Figure 9A ); and a second polyribonucleotide comprising a coding region encoding (in 5' to 3' order): a light chain variable domain (VL) and a CH1 domain (see e.g. Figure 9B ).

[0484] In some embodiments, a CrossMab encoded by one or more polyribonucleotides provided herein is CH1-CLx The antibody agent includes all or part of the 1-18 antibody. In some embodiments, CrossMab CH1-CLxThe antibody agent comprises a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFG DRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof. In some embodiments, CrossMab CH1-CLx The antibody agent comprises a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12). In some embodiments, CrossMab CH1-CLx The antibody agent comprises a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, CrossMab CH1-CLx The antibody agent comprises a light chain variable domain comprising: (i) LCDR1 (QGLDSS H; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21). In some embodiments, CrossMab CH1-CLx The antibody agent comprises (a) a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPH YNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof; and (b) a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, CrossMab CH1-CLxThe antibody agent comprises (a) a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO:6); (ii) HCDR2 (ISPHFARP; SEQ ID NO:9); and (iii) HCDR3 (ARDPFG DRAPHYNYHMDV; SEQ ID NO:12); and (b) a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO:15); (ii) LCDR2 (GTS; SEQ ID NO:18); and (iii) LCDR3 (QRYGG TPIT; SEQ ID NO:21).

[0485] In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx The immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a heavy chain variable domain, and wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDD DTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx The immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a heavy chain variable domain, and wherein the heavy chain variable domain comprises: (i) HC DR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHF ARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMD V; SEQ ID NO: 12). In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx The immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a light chain variable domain, and wherein the light chain variable domain comprises: (i) LC DR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode CrossMa b CH1-CLxThe immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a light chain variable domain, and wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21). In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx Two immunoglobulin chains of an antibody agent: a first immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFG DRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof; and a second immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21); or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx Two immunoglobulin chains of an antibody agent: a first immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises: (i) HCDR1 (DDP YTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); and a second immunoglobulin chain comprising a light chain variable domain, wherein the light chain variable domain comprises: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18); and (iii) LCDR3 (QRYGGTPIT; SEQ ID NO: 21).

[0486] In some embodiments, a CrossMab encoded by one or more polyribonucleotides provided herein is CH1-CLxThe antibody agent comprises a heavy chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 24. In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The antibody agent comprises a light chain variable domain that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence represented by SEQ ID NO: 29. In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein is CH1-CLx The antibody agent comprises a heavy chain variable domain represented by SEQ ID NO: 24. In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The antibody agent comprises a light chain variable domain represented by SEQ ID NO:29.

[0487] In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx The immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a heavy chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the amino acid sequence represented by SEQ ID NO: 24. In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx An immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a light chain variable domain that is at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence represented by SEQ ID NO: 29. In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx The immunoglobulin chain of the antibody agent, wherein the immunoglobulin chain comprises the heavy chain variable domain represented by SEQ ID NO: 24. In some embodiments, the polyribonucleotides described herein encode CrossMab CH1-CLx The immunoglobulin chain of the antibody agent, wherein the immunoglobulin chain comprises a light chain variable domain represented by SEQ ID NO: 29.

[0488] As described above, a CrossMab encoded by one or more polyribonucleotides as described hereinCH1-CLx The antibody agent may comprise one or more heavy chain constant domains. In some embodiments, the one or more heavy chain constant domains comprise a CH3 domain. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH3 domain comprising a G1m3, G1m17, or G1m17,1 allotype. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH3 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the sequence represented by any one of SEQ ID NOs: 68, 71, 74, 77, 80, 83, 86 or 89. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises a CH3 domain having an amino acid sequence represented by any one of SEQ ID NOs: 68, 71, 74, 77, 80, 83, 86, or 89.

[0489] CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent may comprise one or more heavy chain constant domains comprising an amino acid modification (e.g., substitution or deletion) at one or more amino acid positions. For example, a CrossMab encoded by one or more polyribonucleotides as described herein may be a heavy chain constant domain comprising an amino acid modification (e.g., substitution or deletion) at one or more amino acid positions. CH1-CLx Antibody agents may include L / S mutations within the CH3 region (for enhanced FcRn binding) (see Zalevsky J et al. Nat Biotechnol. 2010, incorporated herein by reference). Such mutations are labeled M428L and N434S according to EU numbering and are referred to herein as "LS" or "L / S" (see, e.g., Figure 5C In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises an E294 deletion (for Fc hypersialylation) (see Bas M et al. J Immunol 2019, which is incorporated herein by reference).

[0490] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLxThe antibody agent comprises a CH3 domain comprising one or more of the following mutations: Y349C, T366S, L368A, and Y407V (according to EU numbering). This combination of mutations is referred to herein as "cah." In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises a CH3 domain comprising one or more mutations selected from the group consisting of S354C and T366W (according to EU numbering). This CH3 mutation combination is referred to herein as "cak".

[0491] In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one of the following substitution mutations: M428, N434S, or a combination thereof (e.g., an "L / S" mutation). In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence comprising any one of SEQ ID NOs: 75 and 78. In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one of the following substitution mutations: Y349C, T366S, L368A, and Y407V (according to EU numbering). In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NOs: 81 and 84. In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one of the following substitution mutations: S354C and T366W (according to EU numbering). In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NOs: 87 and 90.

[0492] In some embodiments, the polyribonucleotide encodes an immunoglobulin chain comprising a VH domain operably linked to one or more constant domains, wherein the one or more constant domains comprise a CH3 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NO: 69. In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence encoding a CH3 domain comprising a G1m3, G1m17, or Glm17,1 allotype. In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence comprising any one of SEQ ID NOs: 69 and 72.

[0493] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a light chain constant domain. In some embodiments, the light chain constant domain comprises a kappa light chain constant domain. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described hereinCH1-CLx The antibody agent comprises a kappa light chain constant domain having an amino acid sequence that is at least 80, 85, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 92. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises a kappa light chain constant domain having an amino acid sequence represented by SEQ ID NO: 92. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises a lambda chain variable domain.

[0494] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a hinge domain. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a hinge domain comprising the amino acid sequence represented by SEQ ID NO: 104 (referred to herein as "hinge" in Tables 2 and 4).

[0495] In some embodiments, polyribonucleotide encoding hinge domain. In some embodiments, polyribonucleotide encoding SEQ ID NO: hinge ribonucleic acid sequence represented by 105. In some embodiments, polyribonucleotide encoding hinge domain, described hinge domain includes amino acid modification, and described amino acid modification includes the disappearance of one or more amino acid residues. In some embodiments, polyribonucleotide encoding hinge domain, described hinge domain includes amino acid modification, and described amino acid modification includes conventional Ig hinge domain (represented by SEQ ID NO: 104) missing amino acid residue EPKSC. Such modification is referred to as " hinge _del " or " ΔEPKSC " in this article. In some embodiments, polyribonucleotide encoding SEQ ID NO: hinge ribonucleic acid sequence represented by 111. In some embodiments, polyribonucleotide encoding hinge domain, described hinge domain includes amino acid modification, and described amino acid modification includes C220S mutation (according to EU numbering). Such mutant hinge domain is referred to as " hinge _S " or " C / S " in this article. In some embodiments, the polyribonucleotide encodes the hinge ribonucleic acid sequence represented by SEQ ID NO:108.

[0496] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLxThe antibody agent comprises a CH2 domain. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to that represented by SEQ ID NO: 53. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO:53.

[0497] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH2 domain having one or more mutations (e.g., relative to SEQ ID NO: 53). For example, in some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises one or more of the following mutations: G236A, A330L, and I332E (according to EU numbering). Such a combination of mutations is referred to herein as "GAALIE." Such mutations in the CH2 domain are associated with increased affinity for the Fc receptors FcgRIIA and FcgRIII, thereby enhancing antibody effector function.

[0498] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 56. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO:56.

[0499] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises one or more mutations selected from the group consisting of G236A and I332E (according to EU numbering). Such a CH2 mutation combination is referred to herein as "GAIE". In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLxThe antibody agent comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 59. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO:59.

[0500] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises the mutation: G236A (according to EU numbering), referred to herein as "GA". In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 62. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO: 62.

[0501] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises the mutation: I332E (according to EU numbering), referred to herein as "IE". In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 65. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH2 domain having an amino acid sequence represented by SEQ ID NO:65.

[0502] In some embodiments, the polyribonucleotide encodes an immunoglobulin chain comprising a VH domain operably linked to one or more constant domains, wherein the one or more constant domains comprise a CH2 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NO: 54. In some embodiments, the CH2 ribonucleic acid encodes a CH2 domain having one or more amino acid substitution mutations. For example, in some embodiments, the CH2 ribonucleic acid sequence encodes one or more of the following mutations: G236A, A330L, and I332E (according to EU numbering), referred to herein as "GAALIE". Such mutations in the CH2 domain are associated with increased affinity for Fc receptors FcgRIIA and FcgRIII, thereby enhancing antibody effector function. In some embodiments, the CH2 ribonucleic acid sequence comprises or consists of a sequence according to SEQ ID NO: 57. In some embodiments, the CH2 ribonucleic acid sequence encodes one or more of the following mutations: G236A and I332E (according to EU numbering), referred to herein as "GAIE". In some embodiments, the CH2 ribonucleic acid sequence comprises a sequence according to SEQ ID NO: 60. In some embodiments, the CH2 ribonucleic acid sequence encodes the mutation G236A (according to EU numbering), referred to herein as "GA". In some embodiments, the CH2 ribonucleic acid sequence comprises a sequence according to SEQ ID NO: 63. In some embodiments, the CH2 ribonucleic acid sequence encodes the mutation I332E (according to EU numbering), referred to herein as "IE". In some embodiments, the CH2 ribonucleic acid sequence comprises a sequence according to SEQ ID NO: 66. In some embodiments, the CH2 ribonucleic acid sequence encodes a CH2 domain comprising a deletion E294 (according to EU numbering), referred to herein as "E294del".

[0503] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH1 domain. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH1 domain comprising a G1m3 allotype. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a CH1 domain comprising the G1m17 allotype. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLxThe antibody agent comprises a CH1 domain comprising the amino acids represented by SEQ ID NO: 38 or 41.

[0504] In some embodiments, the polyribonucleotide encodes an immunoglobulin chain comprising a VH domain operably linked to one or more constant domains, wherein the one or more constant domains comprise a CH1 domain. In some embodiments, the polyribonucleotide comprises a CH1 ribonucleic acid sequence according to SEQ ID NO: 39. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising a G1m3 allotype. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising a G1m17 allotype. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence according to SEQ ID NO: 39 or 42.

[0505] In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more mutations. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more added serine residues. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising two additional serine residues added (referred to herein as "SS"). In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 45 or 48. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence represented by SEQ ID NO: 45 or 48. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more charge variation mutations. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more substitution mutations selected from the group consisting of K147E, K213D, or a combination thereof. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% sequence identity to SEQ ID NO: 51 or 866. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence according to SEQ ID NO: 51 or 866.

[0506] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLxThe antibody agent comprises a signal peptide comprising a husec2 signal peptide. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises a signal peptide comprising SEQ ID NO:1.

[0507] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises an immunoglobulin chain encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence represented by SEQ ID NO: 161-208 (heavy chain) and SEQ ID NO: 450 and 451 (light chain). In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises an immunoglobulin chain encoded by a nucleic acid sequence represented by any one of SEQ ID NOs: 161-208 (heavy chain) and SEQ ID NOs: 450 and 451 (light chain).

[0508] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises an immunoglobulin chain encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence represented by SEQ ID NO: 628 (heavy chain) and / or SEQ ID NO: 631 (light chain). In some embodiments, CrossMab CH1-CLx It comprises the immunoglobulin chain represented by SEQ ID NO: 628 (heavy chain) and / or SEQ ID NO: 631 (light chain).

[0509] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLx The antibody agent comprises an immunoglobulin chain (e.g., an immunoglobulin heavy chain) comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence represented by SEQ ID NO: 629. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLxThe antibody agent comprises an immunoglobulin chain (e.g., an immunoglobulin heavy chain) comprising an amino acid sequence represented by any one of SEQ ID NOs: 629. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1-CLx The antibody agent comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) comprising an amino acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 632. In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein is CH1 -CLx The antibody agent comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) comprising the amino acid sequence represented by SEQ ID NO: 632. CrossMab as described herein CH1-CLx Exemplary heavy and light chain configurations of antibody agents are shown in Table 3 below.

[0510] Table 3: Exemplary CrossMabs CH1-CLx Chain Configuration

[0511]

[0512]

[0513]

[0514]

[0515]

[0516]

[0517] C.CrossMab CH1-CLcv

[0518] In some embodiments, the antibody agent as described herein is in a form in which one or more charge variants (cv) are introduced into a domain (e.g., a constant domain, such as a CH1 domain, a CL domain, or a combination thereof). Such forms are referred to herein as "CrossMab CH1-CLcv ” or “CH1-cv”. In some embodiments, such antibody agents comprise charge variants in both arms of the antibody (see, e.g., Figure 4C). Exemplary charge variants are described, for example, in WO2017055539 A1, which is incorporated herein by reference in its entirety. In some embodiments, in contrast to the format described in WO2017055539 A1, the described CH1-cv antibody agents do not include an exchange of the antibody CH1 and CL domains in either arm of the antibody.

[0519] In some embodiments, charge variants (cv) are introduced into the CH1 and / or CL domains of the antibody agent to prevent mispairing of the immunoglobulin chains of the antibody agent. Such charge variants can include, for example, the introduction of one or more positively charged amino acid residues in the CH1 domain and one or more negatively charged amino acid residues in the CL, or vice versa, at specific positions in the CH1 and CL interface.

[0520] In some embodiments, the polyribonucleotide may encode a CrossMab as described herein. CH1-CLcv The heavy and / or light chains of an antibody agent.

[0521] In some embodiments, CrossMab CH1-CLcv The antibody agent can be encoded by two separate polyribonucleotides: a first polyribonucleotide comprising a coding region encoding (in 5' to 3' order): a heavy chain variable domain (VH), a CH1 domain including one or more charge variants as described herein, a hinge region, a CH2 domain, and a CH3 domain (see, e.g., Figure 10A ); and a second polyribonucleotide comprising a coding region encoding (in 5' to 3' order): a light chain variable domain (VL) and a light chain constant domain (CL) comprising one or more charge variants as described herein (see e.g. Figure 10B ).

[0522] In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a first immunoglobulin chain, wherein the first immunoglobulin chain comprises a CL domain, and the CL domain has an amino acid substituted at position 123 (EU numbering) with an amino acid selected from the group consisting of K, R, and H. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a first immunoglobulin chain, wherein the first immunoglobulin chain comprises a CL domain, and the CL domain has an amino acid substituted at position 124 (EU numbering) with an amino acid selected from the group consisting of K, R, and H. In some embodiments, CrossMab CH1-CLcvThe antibody agent comprises a second immunoglobulin chain, wherein the second immunoglobulin chain comprises a CH1 domain having an amino acid substituted at position 147 (EU numbering) with an amino acid selected from E or D (see WO2017055539A1, which is incorporated herein by reference in its entirety).

[0523] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLcv The antibody agent comprises a CH1 domain comprising one or more charge variant mutations. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a CH1 domain, wherein the CH1 domain comprises one or more substitutions including K147E, K213D, or a combination thereof. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a CH1 domain having at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 50 or 865. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a CH1 domain having an amino acid sequence according to SEQ ID NO: 50 or 865. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a CH1 domain comprising a G1m3 allotype. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a CH1 domain comprising the G1m17 allotype.

[0524] In some embodiments, a CrossMab encoded by one or more polyribonucleotides as described herein CH1-CLcv The antibody agent comprises a heavy chain encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to the sequence represented by SEQ ID NO: 209-256. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a heavy chain encoded by a nucleic acid sequence represented by any one of SEQ ID NOs: 209-256. In some embodiments, CrossMab CH1-CLcvThe antibody agent comprises a light chain encoded by a nucleic acid sequence having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% sequence identity to SEQ ID NO: 452 or 453. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a light chain encoded by the nucleic acid sequence represented by SEQ ID NO: 452 or 453.

[0525] In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKYW; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12); or (iv) a combination thereof. In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a heavy chain variable domain comprising: (i) HCDR1 (DDPYTDDDTFTKY W; SEQ ID NO: 6); (ii) HCDR2 (ISPHFARP; SEQ ID NO: 9); and (iii) HCDR3 (ARDPFGDRAPHYNYHMDV; SEQ ID NO: 12). In some embodiments, CrossMab CH1-CLcv The antibody agent comprises a light chain variable domain comprising: (i) LCDR1 (QGLDSSH; SEQ ID NO: 15); (ii) LCDR2 (GTS; SEQ ID NO: 18...

Claims

1. A composition comprising: A first polyribonucleotide encoding an immunoglobulin chain, wherein the immunoglobulin chain comprises a heavy chain variable (VH) domain, the VH domain comprising: (a) a heavy chain complementarity determining region (HCDR) 1 comprising an amino acid sequence according to SEQ ID NO: 6; (b) HCDR2 comprising the amino acid sequence according to SEQ ID NO: 9; and (c) HCDR3 comprising the amino acid sequence according to SEQ ID NO: 12; and A second polyribonucleotide encoding an immunoglobulin chain, wherein the immunoglobulin chain comprises a light chain variable (VL) domain, the VL domain comprising: (a) a light chain complementarity determining region (LCDR) 1 comprising an amino acid sequence according to SEQ ID NO: 15; (b) LCDR2 comprising the amino acid sequence according to SEQ ID NO: 18 (GTS); and (c) LCDR3 comprising the amino acid sequence according to SEQ ID NO:

21.

2. compositions as claimed in claim 1, wherein said first polyribonucleotide comprises VH structural domain coding sequence, and said VH structural domain coding sequence comprises: (a) a HCDR1 coding sequence comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO: 7; (b) a HCDR2 coding sequence comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO: 10; and (c) a HCDR3 coding sequence comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO: 13; and Wherein the second polyribonucleotide comprises a VL domain encoding sequence, and the VL domain encoding sequence comprises: (a) a LCDR1 coding sequence comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO: 16; (b) a LCDR2 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 19 (GGCACCAGC); and (c) a LCDR3 coding sequence comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

22.

3. A composition as claimed in claim 1 or 2, wherein the VH domain comprises or consists of the amino acid sequence according to SEQ ID NO:

24.

4. The composition of any one of claims 1 to 3, wherein the VH domain encoding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 25 or 27.

5. The composition of any one of claims 1-4, wherein the VL domain comprises or consists of the amino acid sequence according to SEQ ID NO:

29.

6. The composition of any one of claims 1-5, wherein the VL domain encoding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO:

30.

7. The composition of any one of claims 1 to 6, wherein: (i) the immunoglobulin chain encoded by the first polyribonucleotide comprises the VH domain, which is operably connected to the CH1 domain, the hinge domain, the CH2 domain and the CH3 domain in sequence, and / or (ii) the immunoglobulin chain encoded by the second polyribonucleotide comprises the VL domain, and the VL domain is operably linked to the CL constant domain.

8. The composition of any one of claims 1 to 6, wherein: (i) the immunoglobulin chain encoded by the first polyribonucleotide comprises the VH domain, which is operably connected to the CL domain, the hinge domain, the CH2 domain and the CH3 domain in sequence, and / or (ii) the immunoglobulin chain encoded by the second polyribonucleotide comprises the VL domain, and the VL domain is operably linked to a CH1 constant domain.

9. The composition of claim 7 or 8, wherein the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO:

53.

10. The compositions of any one of claims 7-9, wherein the first polyribonucleotide comprises a ribonucleic acid sequence, wherein the ribonucleic acid sequence: (i) encodes the CH2 domain, and (ii) comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

54.

11. The composition of any one of claims 7 to 10, wherein the CH3 domain comprises or consists of an amino acid sequence according to SEQ ID NO: 68 or 71.

12. compositions as described in any one among the claim 7-11, wherein said first polyribonucleotide comprises ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes the CH3 domain, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 69 or 72.

13. A composition as described in any one of claims 7 to 12, wherein the hinge domain comprises or consists of an amino acid sequence according to SEQ ID NO: 104 or 110.

14. compositions as described in any one among claim 7-13, wherein said first polyribonucleotide comprises ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes the hinge domain, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 105 or 111.

15. The composition of any one of claims 7-14, wherein the CL domain comprises or consists of the amino acid sequence according to SEQ ID NO:

92.

16. compositions as described in any one among the claim 7-15, wherein said first polyribonucleotide or said second polyribonucleotide comprise ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes the CL domain, and (ii) comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

93.

17. The composition of any one of claims 7-16, wherein the CH1 domain comprises or consists of an amino acid sequence according to SEQ ID NO: 38 or 41.

18. compositions as described in any one among the claim 7-17, wherein said first polyribonucleotide or said second polyribonucleotide comprise ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes the CH1 domain, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 39 or 42.

19. The composition of any one of claims 7-18, wherein the CH1 domain comprises one or more substitution mutations, the one or more substitution mutations comprising or consisting of K147E, K213D, or a combination thereof, and the substitution mutation positions are according to EU numbering.

20. The composition of any one of claims 7-19, wherein the CL domain comprises one or more substitution mutations comprising or consisting of R108A, T109S, E123K, E123R, Q124K, Q124E, Q124R, or a combination thereof, and the substitution mutation positions are according to EU numbering.

21. compositions as described in any one of claims 7-20, wherein the described immunoglobulin chain encoded by the first polyribonucleotide comprises the CL domain, and the described CL domain comprises one or more substitution mutations, and the described one or more substitution mutations comprise Q124E or are composed of it, and the substitution mutation position is according to EU numbering.

22. A composition as described in claim 20 or 21, wherein the immunoglobulin chain encoded by the first polyribonucleotide comprises a CL domain, and the CL domain comprises or consists of an amino acid sequence according to SEQ ID NO:

95.

23. The compositions of any one of claims 20-22, wherein the first polyribonucleotide comprises a ribonucleic acid sequence, wherein the ribonucleic acid sequence: (i) encodes the CL domain, and (ii) comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

96.

24. The composition of any one of claims 7 to 20, wherein the immunoglobulin chain encoded by the first polyribonucleotide comprises a CH1 domain comprising one or more substitution mutations comprising or consisting of K147E and K213D, the immunoglobulin chain encoded by the second polyribonucleotide comprises a CL domain comprising one or more substitution mutations comprising or consisting of E123K and Q124R or E123R and Q124K, and Substitution mutation positions are according to EU numbering.

25. A composition as described in claim 20 or 24, wherein the immunoglobulin chain encoded by the second polyribonucleotide comprises a CL domain, and the CL domain comprises or consists of an amino acid sequence according to SEQ ID NO: 98 or 101.

26. compositions as described in any one in claim 20,24 and 25, wherein said second polyribonucleotide comprises ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes the CL domain, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 99 or 102.

27. The polyribonucleotide of claim 19, 20, 24-26, wherein the immunoglobulin chain encoded by the first polyribonucleotide comprises a CH1 domain, the CH1 domain comprising or consisting of an amino acid sequence according to SEQ ID NO:50 or 865.

28. The composition of any one of claims 19, 20, 24-27, wherein the first polyribonucleotide comprises a ribonucleic acid sequence, wherein the ribonucleic acid sequence: (i) encodes the CH1 domain, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 51 or 866.

29. compositions as described in any one among the claims 1-28, wherein said first and / or second polyribonucleotide comprise ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes a secretion signal, and (ii) comprising a ribonucleic acid sequence according to SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:884 or SEQ ID NO:

887.

30. A composition comprising polyribonucleotides encoding immunoglobulin chains, wherein the immunoglobulin chains comprise single-chain variable fragments (scFv), and the scFv comprises a VH domain, a linker, and a VL domain; wherein the VH domain comprises: (a) a heavy chain complementarity determining region (HCDR) 1 comprising an amino acid sequence according to SEQ ID NO: 6; (b) HCDR2 comprising the amino acid sequence according to SEQ ID NO: 9; and (c) HCDR3 comprising the amino acid sequence according to SEQ ID NO: 12; and wherein the VL domain comprises: (a) a light chain complementarity determining region (LCDR) 1 comprising an amino acid sequence according to SEQ ID NO: 15; (b) LCDR2 comprising the amino acid sequence according to SEQ ID NO: 18 (GTS); and (c) LCDR3 comprising the amino acid sequence according to SEQ ID NO:

21.

31. The composition of claim 30, wherein the scFv comprises, in order: (i) the VH domain comprising or consisting of the amino acid sequence according to SEQ ID NO: 24, (ii) the linker, and (iii) the VL domain comprising or consisting of the amino acid sequence according to SEQ ID NO:

29.

32. polyribonucleotide as claimed in claim 30, wherein said scFv comprises in sequence: (i) the VL domain comprising or consisting of the amino acid sequence according to SEQ ID NO: 29, (ii) the linker, and (iii) the VH domain comprising or consisting of the amino acid sequence according to SEQ ID NO:

24.

33. The composition of any one of claims 30-32, wherein the scFv is operably linked to a hinge domain, a CH2 domain, and a CH3 domain in sequence.

34. A composition as described in any of claims 30-33, wherein the linker comprises an amino acid sequence according to SEQ ID NO: 32 or 35.

35. compositions as described in any one among the claim 30-34, wherein said polyribonucleotide comprises ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encoding the linker, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 33 or 36.

36. A composition as described in any of claims 33-35, wherein the hinge domain comprises or consists of an amino acid sequence according to SEQ ID NO:

107.

37. compositions as described in any one among the claim 33-36, wherein said polyribonucleotide comprises ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes the hinge domain, and (ii) comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

108.

38. The composition of any one of claims 7-29 and 33-37, wherein the CH2 domain comprises one or more substitution mutations comprising or consisting of G236A, A330L, I332E or a combination thereof, and the substitution mutation positions are according to EU numbering.

39. The composition of any one of claims 7-8, 11-29 and 33-38, wherein the CH2 domain comprises or consists of an amino acid sequence according to SEQ ID NO: 56, 59, 62 or 65.

40. The compositions as described in any one of claims 7-8, 11-29 and 33-39, wherein the polyribonucleotides comprise a ribonucleic acid sequence, wherein the ribonucleic acid sequence: (i) encodes the CH2 domain, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 57, 60, 63 or 66.

41. The composition of any one of claims 7-29 and 33-40, wherein the CH3 domain comprises one or more substitution mutations comprising or consisting of Y349C, S354C, T366S, T366W, L368A, Y407V, M428L, N434S or a combination thereof, and the substitution mutation positions are according to EU numbering.

42. The composition of claims 7-29 and 33-41, wherein the CH3 domain comprises one or more substitution mutations, the one or more substitution mutations comprising or consisting of M428L and N434S, and the substitution mutation positions are according to EU numbering.

43. The composition of any one of claims 7-10, 13-29 and 33-42, wherein the CH3 domain comprises or consists of an amino acid sequence according to SEQ ID NO: 74, 77, 80, 83, 86 or 89.

44. as described in any one of claims 7-10,13-29 and 33-43 compositions, wherein said polyribonucleotide comprises ribonucleic acid sequence, said ribonucleic acid sequence: (i) encodes the CH3 domain, and (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 75, 78, 81, 84, 87 or 90.

45. compositions as described in any one among the claim 30-44, wherein said polyribonucleotide comprises ribonucleic acid sequence, and said ribonucleic acid sequence: (i) encodes a secretion signal, and (ii) comprising a ribonucleic acid sequence according to SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:884 or SEQ ID NO:

887.

46. ​​compositions as described in any one in claim 1-7 and 42, wherein the described immunoglobulin chain encoded by the first polyribonucleotide comprises following immunoglobulin chain, and it comprises according to the aminoacid sequence of SEQ ID NO:617 or is composed of it.

47. compositions as described in claim 1-7,42 and 46, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:616 or consists of it.

48. compositions as described in any one in claim 1-7,19 and 42, wherein the described immunoglobulin chain encoded by the first polyribonucleotide comprises following immunoglobulin chain, and it comprises according to the aminoacid sequence of SEQ ID NO:635 or is composed of it.

49. compositions as described in any one in claim 1-7,19,42 and 48, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQ ID NO:634 or is made up of it.

50. compositions as described in any one in claim 1-7,19,41 and 42, wherein the described immunoglobulin chain encoded by the first polyribonucleotide comprises following immunoglobulin chain, and it comprises according to the aminoacid sequence of SEQ ID NO:644 or consists of it.

51. compositions as described in any one in claim 1-7,19,41,42 and 50, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQ ID NO:643 or is made up of it.

52. compositions as described in any one in claim 1-7,19,41 and 42, wherein the described immunoglobulin chain encoded by the first polyribonucleotide comprises following immunoglobulin chain, it comprises according to the aminoacid sequence of SEQ ID NO:641 or is composed of it.

53. compositions as described in any one in claim 1-7,19,41,42 and 52, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQ ID NO:640 or is made up of it.

54. compositions as described in any one in claim 1-7 and 20, wherein the described immunoglobulin chain encoded by the second polyribonucleotide comprises following immunoglobulin chain, and it comprises according to the aminoacid sequence of SEQ ID NO:638 or is composed of it.

55. compositions as described in any one in claim 1-7,20 and 54, wherein said second polyribonucleotide comprises according to the ribonucleic acid sequence of SEQ ID NO:637 or consists of it.

56. compositions as described in any one among the claims 1-7, wherein the described immunoglobulin chain encoded by the second polyribonucleotide comprises following immunoglobulin chain, and it comprises according to the aminoacid sequence of SEQ ID NO:620 or is composed of it.

57. compositions as described in any one in claim 1-7 and 56, wherein said second polyribonucleotide comprises according to the ribonucleic acid sequence of SEQ ID NO:619 or is made up of it.

58. A composition as described in any of claims 30-37, wherein the immunoglobulin chain comprises an immunoglobulin chain comprising or consisting of an amino acid sequence according to SEQ ID NO: 656, 659, 662 or 665.

59. compositions as described in any one among the claims 30-37 and 58, wherein said polyribonucleotide comprises according to SEQ ID NO:655,658,661 or 664 ribonucleic acid sequence or is made up of it.

60. compositions as described in any one among the claims 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:455 or is composed of it.

61. compositions as described in any one among the claims 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:671 or is made up of it.

62. compositions as described in any one among the claim 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:463 or is made up of it.

63. compositions as described in any one among the claim 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:727 or is made up of it.

64. compositions as described in any one among the claim 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:736 or is made up of it.

65. compositions as described in any one among the claims 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:737 or is made up of it.

66. compositions as described in any one among the claim 1-7, wherein said second polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:458 or is made up of it.

67. compositions as described in any one among the claims 1-7, wherein said second polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:467 or is made up of it.

68. compositions as described in any one among the claims 1-7, wherein said second polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:748 or is made up of it.

69. compositions as described in any one among the claims 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:465 or is made up of it.

70. compositions as described in any one among the claims 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:461 or consists of it.

71. compositions as described in any one among claim 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:739 or is made up of it.

72. compositions as described in any one among claim 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:740 or is made up of it.

73. compositions as described in any one among the claim 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:464 or is made up of it.

74. compositions as described in any one among the claims 1-7, wherein said first polyribonucleotide comprises according to the ribonucleic acid sequence of SEQID NO:466 or is made up of it.

75. compositions as claimed in claim 1, wherein said first polyribonucleotide comprises following or is made up of: (a) A polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 617, or (ii) comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 616, and Wherein the second polyribonucleotide comprises or consists of the following: (b) a polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 620, or (ii) comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

619.

76. compositions as claimed in claim 1, wherein said first polyribonucleotide comprises following or is made up of: (a) A polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 623, or (ii) comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 622, and Wherein the second polyribonucleotide comprises or consists of the following: (b) a polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 620, or (ii) comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

619.

77. compositions as claimed in claim 1, wherein said second polyribonucleotide comprises following or consists of: (a) A polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 638 or 668, or (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 637 or 667, Wherein the first polyribonucleotide comprises or consists of the following: (b) a polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 641, or (ii) comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 640; and Wherein the composition further comprises a third polyribonucleotide, wherein the third polyribonucleotide comprises or consists of: (c) a polyribonucleotide comprising: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 644, or (ii) comprising or consisting of the ribonucleic acid sequence according to SEQ ID NO:

643.

78. compositions as claimed in claim 1, wherein said first polyribonucleotide comprises following or is made up of: (a) A polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 635, or (ii) comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 634, and Wherein the second polyribonucleotide comprises or consists of the following: (b) a polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 638 or 668, or (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 637 or 667.

79. compositions as claimed in claim 1, wherein said first polyribonucleotide comprises following or is made up of: (a) A polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 647, or (ii) comprises or consists of the ribonucleic acid sequence according to SEQ ID NO: 646, and Wherein the second polyribonucleotide comprises or consists of the following: (b) a polyribonucleotide, which: (i) encoding an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 638 or 668, or (ii) comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 637 or 667.

80. The composition of claim 1, wherein (a) the first polyribonucleotide comprises SEQ ID NO: 455 or 671 or consists of it, and (b) the second polyribonucleotide comprises or consists of SEQ ID NO:

458.

81. The composition of claim 1, wherein (a) the first polyribonucleotide comprises SEQ ID NO: 461 or consists of it, and (b) the second polyribonucleotide comprises or consists of SEQ ID NO: 467 or 748.

82. The composition of claim 1, wherein (a) the first polyribonucleotide comprises SEQ ID NO: 727 or consists of it, and (b) the second polyribonucleotide comprises or consists of SEQ ID NO: 467 or 748.

83. The composition of claim 1, wherein (a) the first polyribonucleotide comprises SEQ ID NO: 463 or consists of it, and (b) the second polyribonucleotide comprises SEQ ID NO: 467 or 748 or consists of it, and wherein the composition further comprises (c) a third polyribonucleotide, said third polyribonucleotide comprising or consisting of SEQ ID NO:

465.

84. The composition of claim 1, wherein (a) the first polyribonucleotide comprises or consists of SEQ ID NO: 736; and (b) the second polyribonucleotide comprises SEQ ID NO: 467 or 748 or consists of it, and wherein the composition further comprises (c) a third polyribonucleotide, said third polyribonucleotide comprising or consisting of SEQ ID NO:

739.

85. The composition of claim 1, wherein (a) the first polyribonucleotide comprises SEQ ID NO: 737 or consists of it, and (b) the second polyribonucleotide comprises SEQ ID NO: 467 or 748 or consists of it, and wherein the composition further comprises (c) a third polyribonucleotide, said third polyribonucleotide comprising or consisting of SEQ ID NO:

740.

86. compositions as described in any one in claim 1-85, wherein said compositions comprises three or more different polyribonucleotides, wherein each of the three or more different polyribonucleotides encodes an immunoglobulin chain, wherein each of the three or more different polyribonucleotides encodes a different immunoglobulin chain, and Wherein when the immunoglobulin chain is expressed in a cell by the three or more different polyribonucleotides, the immunoglobulin chain forms: (a) at least two different antibody agents; and (b) at most (2x+n+1) / 2 different antibody agents, wherein x is equal to the number of polyribonucleotides encoding immunoglobulin chains in the three or more polyribonucleotides, and n is equal to the number of polyribonucleotides in the three or more polyribonucleotides minus x.

87. The composition of any one of claims 1-86, wherein the composition further comprises lipid nanoparticles, polyplexes (PLX), lipidated polyplexes (LPLX) or liposomes, Wherein the one or more polyribonucleotides are completely or partially encapsulated in the lipid nanoparticle, polyplex (PLX), lipidated polyplex (LPLX) or liposome.

88. The composition of any one of claims 1-87, wherein the composition further comprises lipid nanoparticles, Wherein the polyribonucleotide is encapsulated in the lipid nanoparticle.

89. The composition of claim 87 or 88, wherein each of the lipid nanoparticles comprises: (a) polymer-conjugated lipids; (b) cationic lipids; and (c) one or more neutral lipids.

90. A pharmaceutical composition comprising the composition of any one of claims 1-89 and at least one pharmaceutically acceptable excipient.

91. A method comprising administering to a subject the pharmaceutical composition of claim 90.

92. The pharmaceutical composition of claim 90, for use in treating HIV, said treatment comprising administering said pharmaceutical composition to a subject.

93. The pharmaceutical composition of claim 90, for use in preventing HIV, said prevention comprising administering said pharmaceutical composition to a subject.

94. The method of claim 91 or the pharmaceutical composition for use of claim 92 or 93, wherein administration of the pharmaceutical composition to the subject results in expression in the subject: (a) the immunoglobulin chains of the antibody agent; (b) the antibody agent; or (c) Both.

95. Use of the composition of any one of claims 1-89 or the pharmaceutical composition of claim 90 for treating HIV in a subject.

96. Use of the composition of any one of claims 1-89 or the pharmaceutical composition of claim 90 for preventing HIV in a subject.

97. A method of producing an antibody agent, the method comprising administering the composition of any one of claims 1-89 or the pharmaceutical composition of claim 90 to a cell, such that the cell expresses and secretes the antibody agent.

98. A method comprising the steps of: Determine one or more characteristics of an antibody agent expressed by the composition of any one of claims 1-89 or the pharmaceutical composition of claim 90 introduced into a cell, wherein the one or more characteristics comprise: (i) the protein expression level of the antibody agent; (ii) the binding specificity of the antibody agent to the CD4 binding site of HIV; (iii) the efficacy of the antibody agent in mediating targeted cell death through antibody-dependent cellular cytotoxicity (ADCC); and (iv) the efficacy of the antibody agent in mediating target cell death through complement-dependent cytotoxicity (CDC).

99. A method comprising the steps of: contacting a cell with a composition according to any one of claims 1 to 89 or a pharmaceutical composition according to claim 90 introduced into the cell; and The antibody agent produced by the cell is detected.

100. A manufacturing method, the method comprising the following steps: (a) determining one or more characteristics of the composition of any one of claims 1 to 89 or the pharmaceutical composition of claim 90, said one or more characteristics comprising or consisting of: (i) the length and / or sequence of the polyribonucleotides; (ii) the integrity of the polyribonucleotides; (iii) the presence and / or location of one or more chemical moieties of the polyribonucleotide; (iv) the extent of expression of the antibody agent when the polyribonucleotide is introduced into a cell; (v) the stability of the polyribonucleotide or its combination; (vi) the level of the antibody agent in a biological sample from an organism into which the polyribonucleotide has been introduced; (vii) the binding specificity of the antibody agent expressed by the polyribonucleotide, optionally binding to the CD4 binding site of HIV; (viii) the efficacy of the antibody agent in mediating target cell death via ADCC; (ix) the efficacy of the antibody agent in mediating target cell death via complement dependent cytotoxicity (CDC); (x) the identity and amount / concentration of lipids within the composition; (xi) the size of the lipid nanoparticles within the composition; (xii) the polydispersity of the lipid nanoparticles within the composition; (xiii) the amount / concentration of the polyribonucleotide in the composition; (xiv) the degree of encapsulation of the polyribonucleotide within the lipid nanoparticles; (xv) levels of double-stranded RNA; and (xvi) combinations thereof; (B) comparing the one or more characteristics of the polyribonucleotide to the one or more characteristics of an appropriate reference standard; and (C)(i) if said comparison indicates that said polyribonucleotide or composition thereof meets or exceeds said reference standard, designating said polyribonucleotide or composition thereof for one or more additional steps in manufacture and / or distribution; or (ii) taking an alternative action if the comparison indicates that the polyribonucleotide or combination thereof does not meet or exceed the reference standard.

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