HIV-targeting RNA compositions

Abstract

The present disclosure provides compositions (e.g., pharmaceutical compositions) and related technology (e.g., components thereof and / or methods related thereto) for delivery of anti-HIV antibody agents. The present disclosure provides, among other things, polyribonucleotides encoding immunoglobulin chains of anti-HIV antibody agents. The present disclosure recognizes that HIV mutates rapidly. The highly mutating nature of HIV virions allows them to escape host immune and / or therapeutic pressures.

Description

[Technical Field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 63 / 477,460, filed December 28, 2022, and U.S. Provisional Application No. 63 / 517,542, filed August 3, 2023, which are incorporated by reference in their entireties. [Background technology]

[0002] Human immunodeficiency virus (HIV) is an infectious virus associated with acquired immune deficiency syndrome (AIDS). According to the World Health Organization, over 37 million people worldwide currently have HIV. In 2020, approximately 680,000 people died from HIV-related causes, and approximately 1.5 million people became infected with HIV. Currently, there is no cure for HIV. Summary of the Invention

[0003] The present disclosure recognizes that HIV mutates rapidly. The rapidly mutating nature of HIV virions allows them to escape host immune and / or therapeutic pressures. To combat viral escape, combination therapies targeting HIV, including anti-HIV antibody agents, have been explored. However, the development of such combination therapies has been hampered by several challenges. First, the development of individual HIV therapies is time-consuming and expensive. For example, the development of anti-HIV antibodies presents challenges due to the demanding and expensive production, including purification and formulation methods associated with protein therapeutics. Second, combination therapies present regulatory challenges. In addition to ensuring that combination therapies are safe and effective, the use of combination therapies is complicated by strict regulations regarding the manufacturing of individual therapies, the combination of multiple therapies, and quality control during storage and administration. Third, the administration of antibodies to subjects can be painful and time-consuming. Typically, antibodies are administered intravenously over a longer period of time. Administration of multiple antibodies can increase the complexity of antibody administration, which can result in increased patient discomfort and additional time requirements. Finally, recombinant antibodies can have short serum half-lives.

[0004] The present disclosure provides insights that address these challenges, enabling not only the safe, reliable, and potent delivery of a single anti-HIV antibody agent to a subject, but also the delivery of multiple anti-HIV therapeutic agents, including multiple anti-HIV antibody agents, to a subject. For example, the present disclosure describes an antibody agent or a portion thereof (e.g., an immunoglobulin chain) delivered to a subject via polyribonucleotides. An antibody agent delivered to a subject as one or more polyribonucleotides encoding the antibody agent is referred to herein as a "RiboMab." After delivery of one or more polyribonucleotides encoding the antibody agent to a subject, the antibody agent, or "RiboMab," is expressed by the subject's body. Furthermore, the term "RibobNAb" refers to a RiboMab that includes all or a portion of a broadly neutralizing antibody (bNAb), e.g., a broadly neutralizing antibody that targets HIV. Using polyribonucleotides as therapeutic agents (as opposed to administering the antibody agent itself) requires a simpler and less expensive manufacturing process. The less complex production of polyribonucleotides encoding antibody agent(s) (e.g., anti-HIV antibody agent(s)) can streamline manufacturing (e.g., by avoiding the need for intensive glycan production and characterization), which can result in reduced regulatory and production challenges associated with the development and use of the antibody agent itself. In addition, polyribonucleotides tend to be effective in producing similar effects as recombinant proteins, yet require much smaller amounts to be administered to a subject. This is because, for example, polyribonucleotides encoding anti-HIV antibody agent(s) can be administered to a subject, and the subject's body will produce the anti-HIV antibody agent(s) itself. Using smaller amounts can provide a more comfortable experience for the patient and increase patient compliance with treatment regimens. The present disclosure also provides techniques that address certain limitations of recombinant antibody technology, including, for example, the short serum half-life of recombinant antibodies, by utilizing RNA technology as a format for directly expressing antibody agents in a patient's cells.

[0005] RiboMab technology also enables the simultaneous administration of two or more antibody agents to a subject. For example, antibodies produced by humans typically contain four polypeptide chains: two "heavy" chains and two "light" chains. Each polypeptide chain (whether heavy or light) contains (1) a "variable" domain, whose sequence and structure vary from antibody to antibody and determine the antigen to which the antibody binds, and (2) "constant" domain(s) whose sequence and structure remain largely invariant across a given class of antibody and therefore have little effect on antigen binding. In humans, specialized white blood cells called "B cells" produce antibodies. Heavy and light chains associate to form antibodies through two primary pairings: (1) pairing of the fragment crystallizable (Fc) domains of two heavy chains with each other, and (2) pairing of each of two light chains with a heavy chain via disulfide bonds. During normal antibody production in humans, a single antibody is produced by a single B cell. In this situation, correct pairing of the heavy and light chains is ensured because only one heavy chain and one light chain are present in each B cell. In contrast, administering to a subject a nucleic acid composition encoding more than one antibody agent requires that the immunoglobulin chains (e.g., heavy and light chains) be properly associated to avoid the formation of unwanted by-products (e.g., unintended antibody agents).

[0006] The present disclosure provides, among other things, polyribonucleotides encoding immunoglobulin chains of antibody agents.

[0007] In one aspect, the disclosure provides a polyribonucleotide encoding an immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a single-chain variable fragment (scFv), the scFv comprising a heavy chain variable (VH) domain, a linker, and a light chain variable (VL) domain, wherein the VH domain comprises HCDR1, HCDR2, and HCDR3, wherein HCDR1, HCDR2, and HCDR3 comprise the amino acid sequences according to SEQ ID NOs: 18, 21, and 24, respectively, wherein the VH domain comprises a first mutation at residue 3 and a second mutation at residue 5 compared to the amino acid sequence according to SEQ ID NO: 1494, and the VL domain comprises LCDR1, LCDR2, and LCDR3, wherein LCDR1, LCDR2, and LCDR3 comprise the amino acid sequences according to SEQ ID NOs: 27, 30, and 33, respectively.

[0008] In some embodiments, the first mutation comprises a substitution mutation that results in the presence of a positively charged amino acid at residue 3. In some embodiments, the positively charged amino acid comprises an amino acid selected from Lys (K), Arg (R), or His (H). In some embodiments, the positively charged amino acid comprises a His (H) residue. In some embodiments, the second mutation comprises a substitution mutation that results in the presence of a polar amino acid at residue 5. In some embodiments, the polar amino acid comprises an amino acid selected from Ser (S), Thr (T), Tyr (Y), Asn (N), or Gln (Q). In some embodiments, the polar amino acid comprises a Thr (T) residue. In some embodiments, the VH domain comprises an amino acid sequence at least 85% identical to an amino acid sequence according to SEQ ID NO: 36. In some embodiments, the VH domain comprises an amino acid sequence at least 90% identical to an amino acid sequence according to SEQ ID NO: 36. In some embodiments, the VH domain comprises an amino acid sequence according to SEQ ID NO: 1494 with substitution mutations Q3H and V5T. In some embodiments, the VH domain comprises an amino acid sequence according to SEQ ID NO: 36.

[0009] In some embodiments, the VL domain comprises an amino acid sequence that is at least 85% identical to the amino acid sequence according to SEQ ID NO: 43. In some embodiments, the VL domain comprises an amino acid sequence that is at least 90% identical to the amino acid sequence according to SEQ ID NO: 43. In some embodiments, the VL domain comprises an amino acid sequence according to SEQ ID NO: 43.

[0010] In some embodiments, the scFv comprises, in order: (i) a VH domain, (ii) a linker, and (iii) a VL domain.

[0011] In some embodiments, the scFv comprises, in order: (i) a VL domain, (ii) a linker, and (iii) a VH domain.

[0012] In some embodiments, the linker comprises an amino acid sequence according to SEQ ID NO: 48. In some embodiments, the linker comprises an amino acid sequence according to SEQ ID NO: 59. In some embodiments, the immunoglobulin chain further comprises a second linker following the scFv domain. In some embodiments, the scFv and second linker comprise or consist of an amino acid sequence according to SEQ ID NO: 64, 67, 70, or 73. In some embodiments, the immunoglobulin chain comprises a hinge domain following the second linker. In some embodiments, the hinge domain comprises or consists of an amino acid sequence according to SEQ ID NO: 167.

[0013] In some embodiments, the immunoglobulin chain comprises one or more constant domains, and the scFv is operably linked to the one or more constant domains. In some embodiments, a hinge domain is between the scFv and the one or more constant domains. In some embodiments, the one or more constant domains comprise a CH3 domain. In some embodiments, the CH3 domain comprises a G1m17,1 or G1m3 allotype. In some embodiments, the CH3 domain comprises one or more substitution mutations, the one or more substitution mutations comprising or consisting of M88L, N94S, or a combination thereof, and the positions of the substitution mutations are relative to the amino acid sequence of SEQ ID NO: 1495. In some embodiments, the CH3 domain comprises substitution mutations at residues 16 and 18 compared to the amino acid sequence of SEQ ID NO: 1495. In some embodiments, the CH3 domain comprises an amino acid sequence of SEQ ID NO: 1495 with substitution mutations D16E and L18M. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence of SEQ ID NO: 116.

[0014] In some embodiments, the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1343. In some embodiments, the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1346. In some embodiments, the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1349. In some embodiments, the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1352.

[0015] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a secretion signal. In some embodiments, the secretion signal comprises a ribonucleic acid sequence that is at least 90% identical to SEQ ID NO: 4 or 8. In some embodiments, the secretion signal comprises a ribonucleic acid sequence comprising SEQ ID NO: 4 or 8.

[0016] In some embodiments, the polyribonucleotide comprises one or more non-coding sequence elements. In some embodiments, the one or more non-coding sequence elements improve RNA stability and / or translation efficiency. 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 any combination thereof. In some embodiments, the polyA tail is or comprises a modified polyA sequence, preferably a stuttered polyA tail. In some embodiments, the polyA tail comprises or consists of a sequence at least 90% identical to SEQ ID NO: 16. In some embodiments, the 3' UTR comprises or consists of a nucleic acid sequence at least 90% identical to SEQ ID NO: 14. In some embodiments, the 5' UTR comprises or consists of a nucleic acid sequence at least 90% identical to SEQ ID NO: 10. In some embodiments, the 5' cap is (m27,3'-O)Gppp(m2'-O)ApG. In some embodiments, the polyribonucleotide comprises one or more modified ribonucleotides. In some embodiments, the one or more modified ribonucleotides comprises pseudouridine.

[0017] In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1000. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1048. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1096. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1144.

[0018] In another embodiment, the present disclosure provides a polyribonucleotide comprising or consisting of a ribonucleic acid sequence according to any one of SEQ ID NOs: 1000, 1048, 1096 or 1044.

[0019] The present disclosure also provides, in one aspect, a composition comprising one or more polyribonucleotides described herein. In some embodiments, the composition further comprises a lipid nanoparticle, a polyplex (PLX), a lipid-linked polyplex (LPLX), or a liposome, wherein one or more polyribonucleotides are fully or partially encapsulated in the lipid nanoparticle, the polyplex (PLX), the lipid-linked polyplex (LPLX), or the liposome. In some embodiments, the composition further comprises a lipid nanoparticle, wherein one or more polyribonucleotides are encapsulated in the lipid nanoparticle. In some embodiments, the lipid nanoparticle is a cationic lipid nanoparticle.

[0020] In another aspect, the present disclosure provides a pharmaceutical composition comprising the composition of any one of claims 53 to 56 and at least one pharmaceutically acceptable excipient. In some embodiments, the pharmaceutical composition is used for the treatment and / or prevention of HIV, comprising administering the pharmaceutical composition to a subject.

[0021] In another aspect, the present disclosure provides a method comprising administering to a subject a pharmaceutical composition described herein.

[0022] In some embodiments, administering the pharmaceutical composition to a subject results in expression of the antibody agent in the subject. In some embodiments, the antibody agent is expressed in the subject at a titer of (a) at least 1 μg / ml in plasma, or (b) at least 1 μg / ml in serum. In some embodiments, the antibody agent is expressed in the subject at a titer of (a) at least 10 μg / ml in plasma, or (b) at least 10 μg / ml in serum. In some embodiments, the antibody agent is detectable in the subject's serum for a period of at least 5 days, at least 10 days, at least 15 days, at least 20 days, at least 25 days, or at least 30 days. In some embodiments, the antibody agent is detectable in the subject's serum for a period of at least 30 days.

[0023] In some embodiments, the antibody agent has the ability to neutralize one or more HIV strains when tested in a TZM-bl cell pseudoviral neutralization assay at antibody agent concentrations of up to 25 μg / ml. In some embodiments, the antibody agent delivered as a polyribonucleotide has greater neutralizing activity against one or more HIV strains compared to an equivalent amount of a parental control antibody delivered as a polyribonucleotide, where the parental control antibody is an IgG antibody comprising the same VH and VL domains as the antibody agent. In some embodiments, the one of more HIV strains comprises one or more HIV strains selected from ZM53M.PB12, Du156.12, Q769.d22, 0330.v4.c3, R2184.c04, 89-F1_2_25, CAP204_2_00_F6_6, Ce1176_A3, 6980.v0.c31, PVO.4, CAP45, CNE8, T250-4, 3103.v3.c10, and C1080_c3.

[0024] In some embodiments, the subject has or is at risk of developing an HIV infection.

[0025] In some embodiments, the method is a method of treating and / or preventing HIV infection.

[0026] In some embodiments, the present disclosure also provides the use of a composition described herein or a pharmaceutical composition described herein for the treatment and / or prevention of HIV in a subject.

[0027] In some embodiments, the disclosure provides a method of producing an antibody agent, the method comprising administering a composition described herein or a pharmaceutical composition described herein to a cell, such that the cell expresses and secretes the antibody agent. In some embodiments, the cell is in a subject, and the antibody agent is produced at a therapeutically relevant plasma concentration or therapeutically relevant serum concentration. In some embodiments, the therapeutically relevant plasma concentration or therapeutically relevant serum concentration is at least 10 μg / ml.

[0028] 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 an amino acid sequence according to SEQ ID NO: 18, an HCDR2 comprising an amino acid sequence according to SEQ ID NO: 21, and an HCDR3 comprising an amino acid sequence according to SEQ ID NO: 24.

[0029] 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 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 an amino acid sequence according to SEQ ID NO: 36.

[0030] In some embodiments, the polyribonucleotide comprises a VH domain-encoding sequence. In some embodiments, the VH domain-encoding sequence comprises (a) an HCDR1-encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 19, (b) an HCDR2-encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 22, and (c) an HCDR3-encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 25.

[0031] In some embodiments, the VH domain coding sequence comprises a ribonucleic acid sequence 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: 37. In some embodiments, the VH domain coding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 37. In some embodiments, the VH domain coding sequence comprises a ribonucleic acid sequence 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: 39. In some embodiments, the VH domain coding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 41. In some embodiments, the VH domain coding sequence comprises a ribonucleic acid sequence 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: 39. In some embodiments, the VH domain coding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO:39.

[0032] In some embodiments, an immunoglobulin chain comprising a VH domain described herein comprises one or more constant domains, hi some embodiments, the VH domain is operably linked to one or more constant domains.

[0033] In some embodiments, an immunoglobulin chain comprising a VH domain described herein comprises one or more constant domains, hi some embodiments, the VH domain is operably linked to one or more constant domains.

[0034] In some embodiments, the one or more constant domains comprises a CH2 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain.

[0035] In some embodiments, the one or more constant domains comprises a CH3 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain.

[0036] In some embodiments, the one or more constant domains comprises a hinge domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain.

[0037] In some embodiments, the one or more constant domains comprises a CH1 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain.

[0038] 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.

[0039] In some embodiments, an immunoglobulin chain comprising a VH domain described herein comprises a CH1 domain, a hinge domain, a CH2 domain, and a CH3 domain.

[0040] In some embodiments, an immunoglobulin chain comprising a VH domain described herein comprises a CL domain, a hinge domain, a CH2 domain, and a CH3 domain.

[0041] The present disclosure further provides a polyribonucleotide encoding an immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a variable light (VL) domain.

[0042] In some embodiments, the VL domain comprises (a) a light chain complementarity determining region (LCDR) 1 comprising an amino acid sequence according to SEQ ID NO: 27, (b) an LCDR2 comprising an amino acid sequence according to SEQ ID NO: 30, and (c) an LCDR3 comprising an amino acid sequence according to SEQ ID NO: 33.

[0043] In some embodiments, the polyribonucleotide comprises a VL domain-encoding sequence. In some embodiments, the VL domain-encoding sequence comprises (a) an LCDR1-encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO:28, an LCDR2-encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO:31, and an LCDR3-encoding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO:34.

[0044] In some embodiments, the VL domain comprises or consists of an amino acid sequence according to SEQ ID NO: 43. In some embodiments, the VL domain coding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 44. In some embodiments, the VL domain coding sequence comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 46.

[0045] In some embodiments, the immunoglobulin chain comprising the VL domain further comprises a constant domain. In some embodiments, the VL domain is operably linked to the constant domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the constant domain.

[0046] 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.

[0047] In some embodiments, the immunoglobulin chain comprising a VL domain further comprises a CH1 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain.

[0048] The present disclosure provides, among other things, polyribonucleotides encoding immunoglobulin chains of an antibody agent, wherein the immunoglobulin chains comprise a heavy chain variable (VH) domain and a light chain variable (VL) domain. In some embodiments, the VH domain comprises an HCDR1 comprising an amino acid sequence according to SEQ ID NO: 18, an HCDR2 comprising an amino acid sequence according to SEQ ID NO: 21, and an HCDR3 comprising an amino acid sequence according to SEQ ID NO: 24. In some embodiments, the VL domain comprises an LCDR1 comprising an amino acid sequence according to SEQ ID NO: 27, an LCDR2 comprising an amino acid sequence according to SEQ ID NO: 30, and an LCDR3 comprising an amino acid sequence according to SEQ ID NO: 33.

[0049] In some embodiments, the polyribonucleotide comprises a VH domain coding sequence and a VL domain coding sequence. In some embodiments, the VH domain coding sequence comprises an HCDR1 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 19, an HCDR2 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 22, and an HCDR3 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 25. In some embodiments, the VL domain coding sequence comprises an LCDR1 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 28, an LCDR2 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 31, and an LCDR3 coding sequence comprising or consisting of a ribonucleic acid sequence according to SEQ ID NO: 34.

[0050] In some embodiments, the immunoglobulin chain comprises a single chain variable fragment (scFv), hi some embodiments, the scFv comprises a VH domain, a linker, and a VL domain.

[0051] In some embodiments, the scFv comprises, in order, a VH domain, a linker, and a VL domain, hi some embodiments, the scFv comprises, in order, a VH domain comprising or consisting of an amino acid sequence according to SEQ ID NO: 36, a linker, and a VL domain comprising or consisting of an amino acid sequence according to SEQ ID NO: 43.

[0052] 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: 43, a linker, and a VH domain comprising or consisting of an amino acid sequence according to SEQ ID NO: 36.

[0053] In some embodiments, the linker comprises an amino acid sequence according to SEQ ID NO: 48. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes the linker and that comprises or consists of a sequence according to SEQ ID NO: 49, 51, 53, 55, or 57.

[0054] In some embodiments, the linker comprises an amino acid sequence according to SEQ ID NO: 59. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence that encodes the linker and that comprises or consists of a sequence according to SEQ ID NO: 60 or 62.

[0055] In some embodiments, the immunoglobulin chains comprising the VH and VL domains described herein comprise one or more constant domains, hi some embodiments, the VH and VL domains are operably linked to one or more constant domains.

[0056] In some embodiments, the immunoglobulin chain comprises one or more constant domains, and a hinge domain is between the scFv and the one or more constant domains.

[0057] In some embodiments, the one or more constant domains comprises a CH2 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain.

[0058] In some embodiments, the one or more constant domains comprises a CH3 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain.

[0059] In some embodiments, the one or more constant domains comprises a hinge domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a hinge domain.

[0060] In some embodiments, the one or more constant domains comprises a CH1 domain. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain.

[0061] According to the present disclosure, the CH2 domain of any of the above embodiments may comprise an amino acid sequence 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 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.

[0062] In some embodiments, the ribonucleic acid sequence encoding the CH2 domain comprises a ribonucleic acid sequence 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 ribonucleic acid sequence encoding the CH2 domain comprises or consists of a sequence according to SEQ ID NO:96.

[0063] 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 or consist of G236A, A330L, I332E, or a combination thereof, where the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise or consist of G236A, where the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise or consist of I332E, where the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise or consist of G236A and I332E, where the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CH2 domain comprise or consist of G236A, A330L, and I332E, where the positions of the substitution mutations are based on EU numbering.

[0064] In some embodiments, the CH2 domain comprises an amino acid sequence 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 amino acid sequence of SEQ ID NO: 56. In some embodiments, the CH2 domain comprises or consists of an amino acid sequence of SEQ ID NO: 99. In some embodiments, the CH2 domain comprises an amino acid sequence 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 amino acid sequence of SEQ ID NO: 102. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence of SEQ ID NO: 102.

[0065] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain having a ribonucleic acid sequence 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: 100. 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: 100. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain having a ribonucleic acid sequence 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: 103. 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: 103.

[0066] In some embodiments, the CH2 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 104. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 104.

[0067] 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: 105. 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: 105.

[0068] In some embodiments, the CH2 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 107. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 107.

[0069] 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: 108. 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: 108.

[0070] In some embodiments, the CH2 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 110. In some embodiments, the CH2 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 110.

[0071] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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: 111. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH2 domain and comprising or consisting of a sequence according to SEQ ID NO: 111.

[0072] According to the present disclosure, the CH3 domain of any of the above embodiments can comprise an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 113. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 113.

[0073] 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: 114. 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: 114.

[0074] In some embodiments, the CH3 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 131. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 131.

[0075] 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: 132. 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: 132.

[0076] 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 the positions of the substitution mutations are based on EU numbering.

[0077] In some embodiments, the CH3 domain comprises or consists of an amino acid sequence according to SEQ ID NO: 116. 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: 117.

[0078] In some embodiments, the CH3 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 134. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 134.

[0079] 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: 135. 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: 135.

[0080] 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, where the positions of the substitution mutations are based on 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, where the positions of the substitution mutations are based on 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, where the positions of the substitution mutations are based on EU numbering.

[0081] In some embodiments, the CH3 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 122. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 122.

[0082] 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: 123. 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: 123.

[0083] In some embodiments, the CH3 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 140. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 140.

[0084] 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: 141. 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: 141.

[0085] In some embodiments, the one or more substitution mutations in the CH3 domain comprise or consist of S354C, T366W, or a combination thereof, and the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CH3 domain comprise or consist of S354C, T366W, M428L, N434S, or a combination thereof, and the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CH3 domain comprise or consist of S354C, T366W, M428L, and N434S, and the positions of the substitution mutations are based on EU numbering.

[0086] In some embodiments, the CH3 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 128. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 128.

[0087] 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: 129. 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: 129.

[0088] In some embodiments, the CH3 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 146. In some embodiments, the CH3 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 146.

[0089] 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: 147. 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: 147.

[0090] According to the present disclosure, the hinge domain of any of the above embodiments may comprise an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 161. In some embodiments, the hinge domain comprises or consists of the amino acid sequence according to SEQ ID NO: 161.

[0091] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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: 162. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the hinge domain and comprising or consisting of a sequence according to SEQ ID NO: 162.

[0092] In some embodiments, the hinge domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 164. In some embodiments, the hinge domain comprises or consists of the amino acid sequence according to SEQ ID NO: 164.

[0093] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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: 165. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the hinge domain and comprising or consisting of a sequence according to SEQ ID NO: 165.

[0094] In some embodiments, the hinge domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 167. In some embodiments, the hinge domain comprises or consists of the amino acid sequence according to SEQ ID NO: 167.

[0095] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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: 168. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the hinge domain and comprising or consisting of a sequence according to SEQ ID NO: 168.

[0096] According to the present disclosure, the CH1 domain of any of the above embodiments may comprise an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 76. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 76.

[0097] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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 the ribonucleic acid sequence according to SEQ ID NO: 77. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 77.

[0098] In some embodiments, the CH1 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 81. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 81.

[0099] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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 the ribonucleic acid sequence according to SEQ ID NO: 82. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 82.

[0100] 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 the positions of the substitution mutations are based on EU numbering.

[0101] In some embodiments, the CH1 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 84. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 84.

[0102] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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 the ribonucleic acid sequence according to SEQ ID NO: 85. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 85.

[0103] In some embodiments, the CH1 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 76. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 76.

[0104] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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 the ribonucleic acid sequence according to SEQ ID NO: 77. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 77.

[0105] In some embodiments, the CH1 domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 81. In some embodiments, the CH1 domain comprises or consists of the amino acid sequence according to SEQ ID NO: 81.

[0106] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the 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 the ribonucleic acid sequence according to SEQ ID NO: 82. In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding the CH1 domain and comprising or consisting of a sequence according to SEQ ID NO: 82.

[0107] According to the present disclosure, the CL domain of any of the above embodiments may comprise an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 149. In some embodiments, the CL domain of any of the above embodiments may comprise or consist of an amino acid sequence according to SEQ ID NO: 149.

[0108] 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 a ribonucleic acid sequence according to SEQ ID NO: 150. 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: 150.

[0109] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a light chain constant domain, and 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 Q124E, and the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CL domain comprise or consist of R108A, T109S, or a combination thereof, and the positions of the substitution mutations are based on EU numbering. In some embodiments, the one or more substitution mutations in the CL domain comprise or consist of R108A, T109S, Q124E, or a combination thereof, and the positions of the substitution mutations are based on EU numbering.

[0110] In some embodiments, the CL domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 152. In some embodiments, the CL domain comprises or consists of the amino acid sequence according to SEQ ID NO: 152.

[0111] 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: 153. 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: 153.

[0112] 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, where the positions of the substitution mutations are based on EU numbering.

[0113] In some embodiments, the CL domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 155. In some embodiments, the CL domain comprises or consists of the amino acid sequence according to SEQ ID NO: 155.

[0114] 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: 156. 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: 156.

[0115] In some embodiments, the one or more substitution mutations in the CL domain comprise or consist of E123R, Q124K, or a combination thereof, where the positions of the substitution mutations are based on EU numbering.

[0116] In some embodiments, the CL domain comprises an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 158. In some embodiments, the CL domain comprises or consists of the amino acid sequence according to SEQ ID NO: 158.

[0117] 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: 159. 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: 159.

[0118] In some embodiments, the polyribonucleotides provided herein encode immunoglobulin chains, (i) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH2 domain, wherein 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; (ii) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH3 domain, wherein the CH3 domain comprises one or more substitution mutations, the one or more substitution mutations comprising or consisting of Y349C, S354C, T366S, T366W, L368A, Y407V, M428L, N434S, or a combination thereof; (iii) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CH1 domain, wherein 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; (iv) the polyribonucleotide comprises a ribonucleic acid sequence encoding a CL domain, wherein 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 (v) a combination thereof; The positions of the substitution mutations are based on EU numbering.

[0119] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1307. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1307.

[0120] In some embodiments, the polyribonucleotide 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 a ribonucleic acid sequence according to SEQ ID NO: 1306. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1306.

[0121] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1310. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1310.

[0122] In some embodiments, the polyribonucleotides 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 a ribonucleic acid sequence according to SEQ ID NO: 1309. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1309.

[0123] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1316. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1316.

[0124] In some embodiments, the polyribonucleotides 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 a ribonucleic acid sequence according to SEQ ID NO: 1315. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1315.

[0125] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1325. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1325.

[0126] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1324. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1324.

[0127] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1319. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1319.

[0128] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1318. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1318.

[0129] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1331. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1331.

[0130] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1330. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1330.

[0131] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1334. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1334.

[0132] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1333. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1333.

[0133] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1337. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1337.

[0134] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1336. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1336.

[0135] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1340. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1340.

[0136] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1339. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1339.

[0137] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1313. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1313.

[0138] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1312. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1312.

[0139] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1328. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1328.

[0140] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1327. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1327.

[0141] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1355. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1355.

[0142] In some embodiments, the polyribonucleotides 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 a ribonucleic acid sequence according to SEQ ID NO: 1354. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1354.

[0143] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1322. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of the amino acid sequence according to SEQ ID NO: 1322.

[0144] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1321. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1321.

[0145] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1343. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of a sequence according to SEQ ID NO: 1343.

[0146] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1342. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1342.

[0147] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1346. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of a sequence according to SEQ ID NO: 1346.

[0148] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1345. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1345.

[0149] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1349. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of a sequence according to SEQ ID NO: 1349.

[0150] In some embodiments, the polyribonucleotides 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 according to SEQ ID NO: 1348. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1348.

[0151] In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1352. In some embodiments, the polyribonucleotides provided herein encode an immunoglobulin chain comprising or consisting of a sequence according to SEQ ID NO: 1362.

[0152] In some embodiments, the polyribonucleotides 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 a ribonucleic acid sequence according to SEQ ID NO: 1351. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1351.

[0153] In some embodiments, the polyribonucleotides provided herein encode immunoglobulin chains comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1358, 1361, 1364, 1367, 1370, 1373, 1376, 1379, 1382, 1385, 1388, 1391, 1394, 1397, 1400, 1403, 1406, 1409, 1412, 1415, 1418, 1421, 1424, 1427, 1430, 1433, 1436, 1439, 1442, 1445, 1448, 1451, 1454, 1457, or 1460. In some embodiments, the polyribonucleotides provided herein encode immunoglobulin chains comprising or consisting of a sequence according to SEQ ID NO: 1358, 1361, 1364, 1367, 1370, 1373, 1376, 1379, 1382, 1385, 1388, 1391, 1394, 1397, 1400, 1403, 1406, 1409, 1412, 1415, 1418, 1421, 1424, 1427, 1430, 1433, 1436, 1439, 1442, 1445, 1448, 1451, 1454, 1457, or 1460.

[0154] In some embodiments, the polyribonucleotides 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 a ribonucleic acid sequence according to SEQ ID NO: 1357, 1360, 1363, 1366, 1369, 1372, 1375, 1378, 1381, 1384, 1387, 1390, 1393, 1396, 1399, 1402, 1405, 1408, 1411, 1414, 1417, 1420, 1423, 1426, 1429, 1432, 1435, 1438, 1441, 1444, 1447, 1450, 1453, 1456 or 1459. In some embodiments, the polyribonucleotides provided herein comprise or consist of a ribonucleic acid sequence according to SEQ ID NO: 1357, 1360, 1363, 1366, 1369, 1372, 1375, 1378, 1381, 1384, 1387, 1390, 1393, 1396, 1399, 1402, 1405, 1408, 1411, 1414, 1417, 1420, 1423, 1426, 1429, 1432, 1435, 1438, 1441, 1444, 1447, 1450, 1453, 1456 or 1459.

[0155] In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence encoding a secretion signal.

[0156] In some embodiments, the secretion signal comprises a ribonucleic acid sequence according to SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6 or SEQ ID NO:8.

[0157] In some embodiments, the polyribonucleotide comprises one or more non-coding sequence elements.

[0158] In some embodiments, the one or more non-coding sequence elements improve RNA stability and / or translation efficiency.

[0159] 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.

[0160] In some embodiments, the polyA tail is or comprises a modified polyA sequence, preferably a stuttered polyA tail.

[0161] In some embodiments, the poly-A tail comprises or consists of a sequence that is at least 90%, at least 95%, or at least 99% identical to SEQ ID NO:416.

[0162] 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:14.

[0163] 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:10.

[0164] In some embodiments, the 5' cap is (m27,3'-O)Gppp(m2'-O)ApG.

[0165] In some embodiments, the polyribonucleotide comprises one or more modified ribonucleotides. In some embodiments, the one or more modified ribonucleotides comprises pseudouridine.

[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 a ribonucleic acid sequence according to SEQ ID NO: 850. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 850.

[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 a ribonucleic acid sequence according to SEQ ID NO: 851. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 851.

[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 a ribonucleic acid sequence according to SEQ ID NO: 853. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 853.

[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 a ribonucleic acid sequence according to SEQ ID NO: 900. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 900.

[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 a ribonucleic acid sequence according to SEQ ID NO: 901. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 901.

[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 a ribonucleic acid sequence according to SEQ ID NO: 905. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 905.

[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 SEQ ID NO: 906. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 906.

[0173] 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 SEQ ID NO: 910. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 910.

[0174] 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 SEQ ID NO: 950. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 950.

[0175] 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 SEQ ID NO: 898. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 898.

[0176] 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 SEQ ID NO: 947. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 947.

[0177] 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 SEQ ID NO: 997. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 997.

[0178] 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 SEQ ID NO: 1000. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1000.

[0179] 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 SEQ ID NO: 1048. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1048.

[0180] 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 SEQ ID NO: 1096. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1096.

[0181] 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: 1144. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1144.

[0182] 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: 850-1190. In some embodiments, the polyribonucleotide comprises or consists of a ribonucleic acid sequence according to any one of SEQ ID NOs: 850-1190.

[0183] In some embodiments, the polyribonucleotide is a non-naturally occurring polyribonucleotide.

[0184] In some embodiments, the polyribonucleotide is an engineered polyribonucleotide.

[0185] In some embodiments, the polyribonucleotide is an isolated polyribonucleotide.

[0186] The present disclosure provides, among other things, compositions comprising one or more polyribonucleotides described herein.

[0187] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1307 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1313.

[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 NOs: 1306, 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 NOs: 1312.

[0189] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1310 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1313.

[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 NOs: 1309, 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 NOs: 1312.

[0191] In some embodiments, the composition comprises or consists of polyribonucleotides encoding immunoglobulin chains comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1316 and polyribonucleotides encoding immunoglobulin chains comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1313.

[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 NOs: 1315, 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 NOs: 1312.

[0193] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1319 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1322.

[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 NOs: 1318, 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 NOs: 1321.

[0195] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1331 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1322.

[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 NOs: 1330, 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 NOs: 1321.

[0197] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1334 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1322.

[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 NOs: 1333, 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 NOs: 1321.

[0199] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1337 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1322.

[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 NOs: 1336, 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 NOs: 1321.

[0201] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1340 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1322.

[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 NOs: 1339, 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 NOs: 1321.

[0203] In some embodiments, the composition comprises or consists of a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1325 and a polyribonucleotide encoding an immunoglobulin chain comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1328.

[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 NOs: 1324, 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 NOs: 1327.

[0205] In some embodiments, the composition comprises or consists of polyribonucleotides encoding immunoglobulin chains comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1325 and polyribonucleotides encoding immunoglobulin chains comprising an amino acid sequence 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 amino acid sequence according to SEQ ID NO: 1355.

[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 NOs: 1324, 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 NOs: 1354.

[0207] 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 liposome.

[0208] In some embodiments, the composition further comprises lipid nanoparticles, wherein the one or more polyribonucleotides are encapsulated within the lipid nanoparticles.

[0209] In some embodiments, the lipid nanoparticles are targeted to hepatocytes.

[0210] In some embodiments, the lipid nanoparticles are targeted to secondary lymphoid organ cells.

[0211] In some embodiments, the lipid nanoparticles are targeted to lung cells.

[0212] In some embodiments, the lipid nanoparticles are cationic lipid nanoparticles.

[0213] In some embodiments, the lipid nanoparticles each comprise a polymer-bound lipid, a cationic lipid, and one or more neutral lipids.

[0214] In some embodiments, the polymer-conjugated lipid comprises a PEG-conjugated lipid.

[0215] In some embodiments, the polymer-conjugated lipid comprises 2-[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide.

[0216] In some embodiments, the one or more neutral lipids comprise 1,2-distearoyl-sn-glycero-3-phosphocholine (DPSC).

[0217] In some embodiments, the one or more neutral lipids comprise cholesterol.

[0218] In some embodiments, the cationic lipid comprises ((3-hydroxypropyl)azanediyl)bis(nonane-9,1-diyl)bis(2-butyloctanoate).

[0219] In some embodiments, the lipid nanoparticles each comprise 2-[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide, DPSC, cholesterol, and ((3-hydroxypropyl)azanediyl)bis(nonane-9,1-diyl)bis(2-butyloctanoate).

[0220] In some embodiments, the lipid nanoparticles comprise polymer-bound lipids at about 1-2.5 mol% of the total lipids, cationic lipids at 35-65 mol% of the total lipids, and one or more neutral lipids present at 35-65 mol% of the total lipids.

[0221] In some embodiments, the lipid nanoparticles have an average diameter of about 50-150 nm.

[0222] The present disclosure also provides pharmaceutical compositions, in some embodiments, comprising a composition provided herein and at least one pharmaceutically acceptable excipient.

[0223] In some embodiments, the pharmaceutical agent comprises a cryoprotectant. In some embodiments, the pharmaceutical agent comprises an aqueous buffer solution.

[0224] The present disclosure provides, among other things, a method.

[0225] In some embodiments, the methods include administering to a subject a pharmaceutical composition provided herein.

[0226] In some embodiments, the pharmaceutical compositions provided herein are for use in the treatment of HIV, comprising administering the pharmaceutical composition to a subject.

[0227] In some embodiments, the pharmaceutical compositions provided herein are for use in preventing HIV, comprising administering the pharmaceutical composition to a subject.

[0228] In some embodiments, the methods or pharmaceutical compositions for uses provided herein comprise administering the pharmaceutical composition to a subject, whereby the subject expresses an immunoglobulin chain of the antibody agent, the antibody agent, or both.

[0229] In some embodiments, the immunoglobulin chains of the antibody agent, the antibody agent, or both, are expressed in the subject at a titer of at least 1 μg / ml in plasma or serum.

[0230] In some embodiments, the antibody agent exhibits a geometric mean IC50 of less than 0.3 μg / ml for five neutralizing strains against the neutralizing strains of the International Reference Panel when tested in a TZM-bl cell pseudovirus neutralization assay at antibody agent concentrations up to 25 μg / ml.

[0231] In some embodiments, the antibody agent has the ability to neutralize one or more HIV strains when tested in a TZM-bl cell pseudoviral neutralization assay at antibody agent concentrations up to 25 μg / ml.

[0232] In some embodiments, the antibody agent has the ability to neutralize one or more HIV strains at a level within three-fold of the level of an equivalent amount of a recombinant reference antibody.

[0233] In some embodiments, the recombinant reference antibody is an unmodified wild-type IgG antibody that comprises the same HCDR1, HCDR2, HCDR2, LCDR1, LCDR2, and LCDR3 as the antibody agent.

[0234] 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 once a week. In some embodiments, one or more doses of the pharmaceutical composition are administered to the subject twice a week.

[0235] 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.

[0236] In some embodiments, the subject has or is at risk of developing an HIV infection.

[0237] In some embodiments, the method is a method of treating an HIV infection.

[0238] In some embodiments, the method is a method of preventing HIV infection.

[0239] Also provided herein are uses of the polyribonucleotides, compositions and pharmaceutical compositions described herein.

[0240] In some embodiments, there is provided a use of a composition or pharmaceutical composition provided herein for the treatment of HIV in a subject.

[0241] In some embodiments, there is provided a use of a composition or pharmaceutical composition provided herein for the prevention of HIV in a subject.

[0242] In some embodiments, the subject has or is at risk of developing an HIV infection.

[0243] Additionally, the present disclosure provides methods for producing an antibody agent. In some embodiments, the methods include administering a composition or pharmaceutical composition provided herein to a cell, such that the cell expresses and secretes the antibody agent.

[0244] In some embodiments, the cells are hepatocytes.

[0245] In some embodiments, the cell is in a subject.

[0246] In some embodiments, the cells are ex vivo cells.

[0247] In some embodiments, the antibody agent is produced at a therapeutically relevant plasma concentration or therapeutically relevant serum concentration, hi some embodiments, the therapeutically relevant plasma concentration or therapeutically relevant serum concentration is at least 1 μg / ml.

[0248] The present disclosure further provides methods for determining one or more characteristics of an antibody agent expressed from a polyribonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, a polyribonucleotide, composition, or pharmaceutical composition provided herein is introduced into a cell. In some embodiments, the one or more characteristics include (i) the protein expression level of the antibody agent; (ii) the binding specificity of the antibody agent to the HIV head-end binding site; (iii) the efficacy of the antibody agent in mediating target cell death by antibody-dependent cellular cytotoxicity (ADCC); and (iv) the efficacy of the antibody agent in mediating target cell death by complement-dependent cytotoxicity (CDC).

[0249] The present disclosure further provides methods comprising contacting a cell with a polyribonucleotide, composition, or pharmaceutical composition provided herein. In some embodiments, the method further comprises detecting an antibody agent produced by the cell.

[0250] In some embodiments, the cells are hepatocytes.

[0251] In some embodiments, the determining step comprises comparing one or more characteristics of the antibody agent to the one or more characteristics of a reference antibody that specifically binds to the head-end binding site of HIV.

[0252] In some embodiments, the determining step comprises assessing a protein expression level of the antibody agent above a threshold level.

[0253] In some embodiments, the threshold level is a level sufficient to induce ADCC.

[0254] In some embodiments, the determining step comprises assessing binding of the antibody agent to a blunt end (eg, V1V2 region) binding site of HIV.

[0255] In some embodiments, the determining step comprises evaluating the antibody agent in a TZM-bl cell pseudoviral neutralization assay at antibody agent concentrations of up to 25 μg / ml.

[0256] In some embodiments, the cell is present in a subject.

[0257] In some embodiments, the cells are ex vivo cells.

[0258] In some embodiments, the one or more characteristics include antibody levels in one or more tissues in the subject.

[0259] The present disclosure also provides methods of manufacturing. In some embodiments, the methods include: (A) determining one or more characteristics of a polyribonucleotide, composition, or pharmaceutical composition provided herein, wherein the one or more characteristics are: (i) the length and / or sequence of the polyribonucleotide; (ii) polyribonucleotide integrity; (iii) the presence and / or location of one or more chemical moieties in the polyribonucleotide; (iv) the degree of expression of the antibody agent when polyribonucleotides are introduced into cells; (v) stability of polyribonucleotides or compositions thereof; (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 an antibody agent expressed from the polyribonucleotide, optionally binding specificity for the CD4 binding site of HIV; (viii) the efficacy of the antibody agent in mediating target cell death by ADCC; (ix) the efficacy of the antibody agent in mediating target cell death by complement-dependent cytotoxicity (CDC); (x) the identity and amount / concentration of lipids in the composition; (xi) particle size of the lipid nanoparticles in the composition; (xii) the polydispersity of the lipid nanoparticles in the composition; (xiii) the amount / concentration of polyribonucleotides in the composition; (xiv) the extent to which polyribonucleotides are encapsulated within lipid nanoparticles; (xv) levels of double-stranded RNA; and (xvi) combinations thereof the determination comprising or consisting of; (B) comparing one or more characteristics of the polyribonucleotide with the one or more characteristics of a suitable reference standard; and (C)(i) designating the polyribonucleotide or composition thereof for one or more further steps of manufacture and / or sale if the comparison indicates that the polyribonucleotide or composition thereof meets or exceeds the reference standard; or (C)(ii) Taking other action if the comparison indicates that the polyribonucleotide or composition thereof does not meet or exceed the reference standard. Includes:

[0260] In some embodiments, polyribonucleotides are evaluated and the one or more further steps of step (C)(i) are or include at least formulation of the polyribonucleotides.

[0261] In some embodiments, the composition or pharmaceutical composition is evaluated and the one or more further steps of step (C)(i) are or include the release and sale of the composition or pharmaceutical composition.

[0262] Thus, the present disclosure provides a technology that allows for the expression of multiple anti-HIV antibodies in a subject, thereby increasing the broadness and potency of anti-HIV antibodies simultaneously present in the subject and reducing the possibility of viral escape.

[0263] The provided technology is described in more detail herein, including exemplary polyribonucleotides, compositions comprising such polyribonucleotides, and methods of making and using such polyribonucleotides. [Brief explanation of the drawings]

[0264] [Figure 1] Schematic diagram of the HIV genome (Panel A) and the structure of the HIV virion particle (Panel B) are shown. Figure adapted from Musumeci et al., 2015 Molecules, which is incorporated herein by reference in its entirety. [Figure 2] 1 shows potential epitope regions on HIV virion particles to which antibody agents (e.g., broadly neutralizing antibodies (bNAbs) or variants thereof) can bind. Diagram adapted from McCoy and Burton, 2017 Immunol Rev., incorporated herein by reference in its entirety. [Figure 3] 1 illustrates an exemplary therapeutic strategy utilizing the RiboMab technology described herein for the delivery and expression of anti-HIV RibobNAbs. [Figure 4] 1 shows exemplary formats of the PGDM1400 RibobNab described herein, which may include IgG (Panel A), CrossMabCH1-CLx (Panel B), CrossMabCH1-CLv (Panel D), or various orientations / linkers of scFv-Fc RibobNAb (Panels C and E). [Figure 5]Figure 1 shows exemplary Fc-modified forms of the PGDM1400 RibobNab described herein. Exemplary RiboMab formats can include an unmodified Fc domain (Panel A) or modified forms, including GAALIE / GAIE / GA / IE (Panel B), L / S (Panel D), and / or knob-into-hole RiboMab (Panel C), as shown in Panels B-D. [Figure 6] Schematic representation of an exemplary polyribonucleotide encoding the heavy chain (Panel A) and light chain (Panel B) of an exemplary PGDM1400 IgG1 antibody agent. [Figure 7] Figure 1 shows the % broadness and efficacy of various broadly neutralizing antibodies, including PGDM1400 against HIV, when tested against a panel of 109 pseudoviruses. [Figure 8] Schematic diagrams of exemplary polyribonucleotides encoding scFv-Fc PGDM1400 antibody agents PGDM1400 scFV-Fc VH-LL4 / 5-VL (panel A) and PGDM1400 scFV-Fc VL-LL4 / 5-VH (panel B) are shown. [Figure 9] Schematic diagrams of exemplary polyribonucleotides encoding the heavy chain (Panel A) and light chain (Panel B) of an exemplary PGDM1400 CrossMabCH1-CLx antibody agent are shown. [Figure 10] Schematic diagrams of exemplary polyribonucleotides encoding the heavy chain (Panel A) and light chain (Panel B) of an exemplary PGDM1400 CrossMabCH1-CLcv antibody agent are shown. [Figure 11] 1 shows exemplary concentrations of PGDM1400 and PGDM1400 L / S RibobNAb compared to a control RiboMab as determined by Gyros ELISA according to Example 5. [Figure 12] 1 shows an exemplary Western blot analysis of PGDM1400 and PGDM1400 L / S RibobNAb compared to a control RiboMab under non-reducing conditions according to Example 5. [Figure 13]Exemplary concentrations of scFv-Fc PGDM1400 L / S RibobNAbs (e.g., VH-LL4-VL, VL-LL5-VH, VH-LL5-VL, and VL-LL4-VH) are shown compared to IgG control RiboMab and parental IgG as determined by Gyros ELISA according to Example 6. [Figure 14] An exemplary Western blot analysis of scFv-Fc PGDM1400 L / S RibobNAbs (e.g., VH-LL4-VL, VL-LL5-VH, VH-LL5-VL, and VL-LL4-VH) under non-reducing conditions from Example 6 is shown compared to control RiboMab and parental IgG. [Figure 15] Exemplary concentrations of CrossMab PGDM1400 L / S RibobNAbs (e.g., CrossMabCH1-CLcv, and CrossMabCH1-CLx) as determined by Gyros ELISA according to Example 7 are shown in comparison to control RiboMabs and parental Abs. [Figure 16] 1 shows an exemplary Western blot analysis of CrossMab PGDM1400 L / S RibobNAbs (e.g., CrossMabCH1-CLcv, and CrossMabCH1-CLx) according to Example 7 compared to control RiboMabs and parental Abs. [Figure 17] Exemplary pharmacokinetic (PK) profiles of PGDM1400 IgG and PGDM1400 IgG L / S RibobNAb compared to control RiboMab are shown in NSG mice (Panel A) and NSG Tg32 mice (Panel B) as determined by Gyros ELISA according to Example 8. Quantitation of PGDM1400 RibobNAb was performed using serum obtained from NSG and hFcRn NSG Tg32 mice. In vivo concentrations in μg / mL are shown on a logarithmic scale on the y-axis. The x-axis indicates the time of each blood sample in days. [Figure 18]Figure 1 shows exemplary pharmacokinetic (PK) profiles of PGDM1400 IgG, IgG L / S, and scFv-Fc L / S RibobNAb. Quantitation of PGDM1400 RibobNAb was performed using serum obtained from hFcRn NSG Tg32 mice. In vivo concentrations, expressed in μg / mL, are shown on a logarithmic scale on the y-axis. The x-axis indicates the time of each blood sample in days. Results for PGDM1400 VL-LL5-VH L / S scFv-Fc as determined by Gyros ELISA according to Example 12 are shown relative to PGDM1400 IgG and PGDM1400 L / S IgG RibobNAb. Two different doses of PGDM1400 VL-LL5-VH L / S scFv-Fc (30 μg and 19.56 μg) and PGDM1400 L / S IgG (30 μg and 10 μg) were analyzed. [Figure 19] Exemplary results from a pseudovirus virus neutralization test (pVNT) are shown in which TZM.b1 cells were exposed to PGDM1400 scFv-Fc L / S (VH-LL4-VL, VL-LL5-VH, VH-LL5-VL, and VL-LL4-VH configurations) and the pseudoviruses ZM53M.PB12, Du156.12, Q769.d22, 0330.v4.c3, R2184.c04, and 89-F1_2_25. Murine leukemia virus (MuLV) pseudovirus was used as a negative control. Results are expressed as IC50 and IC80 values, or antibody / IgG concentrations resulting in a 50% and 80% reduction in relative luminescence units (RLU) compared to untreated virus control wells. [Figure 20]Exemplary results from pVNT were shown in which TZM.b1 cells were exposed to PGDM1400 scFv-Fc L / S (VH-LL4-VL, VH-LL5-VL, VL-LL4-VH, and VL-LL5-VH configurations) and the pseudoviruses CAP304_2_00_F6_6, Ce1176_A3, 6980.v0.c31, 1012_11_TC21_3257, and PVO.4. Murine leukemia virus (MuLV) pseudovirus was used as a negative control. Results are expressed as IC50 and IC80 values, or antibody / IgG concentrations resulting in a 50% and 80% reduction in relative luminescence units (RLU) compared to untreated virus control wells. [Figure 21] Exemplary results are shown for pVNT in which TZM.b1 cells were exposed to PGDM1400 CrossMabCH1-CLx L / S and PGDM1400 CrossMabCH1-CHcv L / S RibobNAbs and the pseudoviruses ZM53M.PB12, Du156.12, Q769.d22, 0330.v4.c3, R2184.c04, and 89-F1_2_25. Murine leukemia virus (MuLV) pseudovirus was used as a negative control. Results are expressed as IC50 and IC80 values, or antibody / IgG concentrations resulting in a 50% and 80% reduction in relative luminescence units (RLU) compared to untreated virus control wells. [Figure 22] Exemplary results from pVNT were shown in which TZM.b1 cells were exposed to PGDM1400 scFv-Fc(VL-LL5-VH)L / S RibobNAb and the pseudoviruses CAP45, X2088_c9, CNE8, T250-4, QH0692.42, and 3103.v3.c10 (Panel A), and C1080_c3, T278-50, ZM109F.PB4, and Du156.12 (Panel B). Murine leukemia virus (MuLV) pseudovirus was used as a negative control. Results are expressed as IC50 and IC80 values, or antibody / IgG concentrations resulting in a 50% and 80% reduction in relative luminescence units (RLU) compared to untreated virus control wells. DETAILED DESCRIPTION OF THE INVENTION

[0265] definition The compounds of the present disclosure include those outlined above and are further exemplified by the classes, subclasses, and species disclosed herein. As used herein, the following definitions shall apply unless otherwise specified. For purposes of this disclosure, chemical elements are identified according to the Periodic Table of Elements, CAS version, Handbook of Chemistry and Physics, 75th Ed. Furthermore, 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 Ed., Ed.: Smith, M.B. and March, J., John Wiley & Sons, New York: 2001, the contents of which are incorporated herein by reference in their entireties.

[0266] Unless otherwise specified, structures depicted herein are intended to include all stereoisomeric (e.g., enantiomeric or diastereomeric) forms of the structure, as well as all geometric or conformational isomeric forms of the structure. For example, R and S configurations of each stereocenter are contemplated as part of this disclosure. Accordingly, single stereochemical isomers as well as enantiomeric, diastereomeric, and geometric (or conformational) mixtures of the provided compounds are within the scope of this disclosure. For example, in some instances, the provided compounds depict one or more stereoisomers of the compound, which, unless otherwise specified, represent each stereoisomer individually and / or in mixtures. Unless otherwise specified, all tautomeric forms of the provided compounds are within the scope of this disclosure.

[0267] Unless otherwise stated, structures depicted herein are meant to include compounds that differ only in the presence of one or more isotopically enriched atoms. For example, compounds having the present structures including the replacement of a hydrogen by deuterium or tritium, or the replacement of a carbon by a C- or C-enriched carbon are within the scope of this disclosure.

[0268] About: The term "about," when used herein with respect to a value, refers to a value that is similar in relationship to the referenced value. Generally, a person of ordinary skill in the art familiar with the context will recognize the appropriate 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.

[0269] Drug: As used herein, the term "drug" may refer to a physical entity. In some embodiments, a drug may be characterized by a particular feature and / or effect. For example, as used herein, the term "therapeutic agent" refers to a physical entity that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect. In some embodiments, a drug may be any chemical class of compound, molecule, or entity, including, for example, a small molecule, polypeptide, nucleic acid, sugar, lipid, metal, or combination or complex thereof.

[0270] 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 units of unsaturation, having a single point of attachment to the rest of the molecule, or a monocyclic or bicyclic hydrocarbon that is fully saturated or contains one or more units of unsaturation, but is not aromatic (also referred to herein as "cycloaliphatic"). Unless otherwise specified, aliphatic groups contain 1-12 aliphatic carbon atoms. In some embodiments, aliphatic groups contain 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 still other embodiments, aliphatic groups contain 1-3 aliphatic carbon atoms (e.g., C 1-3 ), and in yet other embodiments, aliphatic groups contain 1-2 aliphatic carbon atoms (e.g., C 1-2 Suitable aliphatic groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, or alkynyl groups, and hybrids thereof. Preferred aliphatic groups include C 1-6 It is alkyl.

[0271] Alkyl: The term “alkyl” used alone or as part of a larger moiety (unless otherwise specified) refers to an alkyl group having 1 to 12, 1 to 10, 1 to 8, 1 to 6, 1 to 4, 1 to 3, or 1 to 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 ) refers to a saturated, optionally substituted, straight or branched chain hydrocarbon group having a carbon atom. Exemplary alkyl groups include methyl, ethyl, propyl, butyl, pentyl, hexyl, and heptyl.

[0272] Alkylene: The term "alkylene" refers to a divalent alkyl group. In some embodiments, an "alkylene" is a divalent straight or branched alkyl group. In some embodiments, an "alkylene chain" is a polymethylene group, i.e., -(CH) n -, where n is a positive integer, e.g., 1 to 6, 1 to 4, 1 to 3, 1 to 2, or 2 to 3. An optionally substituted alkylene chain is a polymethylene group in which one or more methylene hydrogen atoms are optionally replaced with a substituent. Suitable substituents may include those described below for substituted aliphatic groups and also include those described herein. It will be recognized that two substituents on an alkylene group may be joined together to form a ring system. In certain embodiments, two substituents may be joined to form a 3- to 7-membered ring. The substituents may be on the same or different atoms. The suffix "-ene" or "-enyl," when added to certain groups herein, is intended to refer to a difunctional portion of said group. For example, "-ene" or "-enyl," when added to "cyclopropyl," results in "cyclopropylene" or "cyclopropylenyl," and includes difunctional cyclopropyl groups, e.g., [ka] is intended to refer to.

[0273] Alkenyl: The term “alkenyl,” used alone or as part of a larger moiety, refers to an alkyl group having at least one double bond and (unless otherwise specified) 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (e.g., C 2-12 , C 2-10 , C 2-8 , C 2-6 , C 2-4 or C 2-3The term "cycloalkenyl" refers to an optionally substituted straight-chain, branched-chain, or cyclic hydrocarbon group having at least one carbon-carbon double bond and having from about 3 to about 10 carbon atoms. Exemplary alkenyl groups include ethenyl, propenyl, butenyl, pentenyl, hexenyl, and heptenyl. The term "cycloalkenyl" refers to an optionally substituted non-aromatic monocyclic or multicyclic 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.

[0274] Alkynyl: The term “alkynyl,” used alone or as part of a larger moiety, refers to an alkynyl group having at least one triple bond and (unless otherwise specified) 2 to 12, 2 to 10, 2 to 8, 2 to 6, 2 to 4, or 2 to 3 carbon atoms (e.g., C 2-12 , C 2-10 , C 2-8 , C 2-6 , C 2-4 or C 2-3 ) refers to an optionally substituted straight or branched chain hydrocarbon group having an alkynyl group. Exemplary alkynyl groups include ethynyl, propynyl, butynyl, pentynyl, hexynyl, and heptynyl.

[0275] Amino acid: As used herein, the term "amino acid" in its broadest sense refers to a compound and / or substance that can be, is, or has been incorporated into a polypeptide chain, for example, by the formation of one or more peptide bonds. In some embodiments, an amino acid has the general structure HN-C(H)(R)-COOH. In some embodiments, an amino acid is a naturally occurring amino acid. In some embodiments, an amino acid is a non-natural amino acid, in some embodiments, an amino acid is a D-amino acid, and in some embodiments, an amino acid is an L-amino acid. A "standard amino acid" refers to any of the 20 standard L-amino acids commonly found in naturally occurring peptides. A "non-standard amino acid" refers to any amino acid other than the standard amino acids, whether prepared synthetically or obtained from a natural source. In some embodiments, amino acids, including the carboxy- and / or amino-terminal amino acids in a polypeptide, may contain structural modifications compared to the general structures above. For example, in some embodiments, an amino acid may be modified relative to the general structure by methylation, amidation, acetylation, pegylation, glycosylation, phosphorylation, and / or substitution (e.g., of an amino group, a carboxylic acid group, one or more protons, and / or a hydroxyl group). In some embodiments, such modifications may, for example, alter the circulating half-life of a polypeptide containing the modified amino acid compared to one containing the otherwise identical unmodified amino acid. In some embodiments, such modifications do not significantly alter the relevant activity of a polypeptide containing the modified amino acid compared to one containing the otherwise identical unmodified amino acid. As will be clear from the context, the term "amino acid" may, in some embodiments, be used to refer to a free amino acid, and in some embodiments, to an amino acid residue of a polypeptide.

[0276] Antibody Agent: As used herein, the term "antibody agent" refers to a polypeptide or polypeptide complex that contains sufficient immunoglobulin structural elements to confer specific binding to a particular antigen. Exemplary antibody agents include, but are not limited to, monoclonal or polyclonal antibodies. In some embodiments, an antibody agent may contain one or more constant region sequences characteristic of murine, rabbit, primate, or human antibodies. In some embodiments, an antibody agent may contain one or more sequence elements that have been humanized, primatized, chimerized, etc., as known in the art. In some embodiments, the term "antibody agent" is used to refer to one or more constructs or formats known or developed in the art to utilize the structural and functional characteristics of antibodies in alternative presentations. For example, in some embodiments, antibody agents utilized in accordance with the present disclosure include, but are not limited to, unmodified IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies (e.g., Zybodies®); CrossMabs (e.g., CrossMabs®); CH1-CL , CrossMab CH1-CLcv Bispecific CrossMAb with Knobs-in-Hole CH1-CLantibody fragments, e.g., Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments, and isolated complementarity determining regions (CDRs) or sets thereof; single chain Fv (scFv); scFv-Fc fusions; polypeptide-Fc fusions; single domain antibodies (e.g., shark single domain antibodies, e.g., IgNAR or fragments thereof); camelid antibodies; masked antibodies (e.g., Probody®), small modular immunopharmaceuticals ("SMIP™"); single chain or tandem diabodies (TandAb®) ); VHH; Anticalin®; Nanobody® minibody; BiTE®; Ankyrin repeat protein or DARPIN®; Avimer®; DART; TCR-like antibody; Adnectin®; Affilin®; Transbody®; Affibody®; TrimerX®; Microprotein; Fynomer®, Centilin®; and KALBITOR®. In some embodiments, chains and / or fragments of such antibodies and fragments may be used in combination, e.g., scFv-Fc arms may be used in combination with traditional antibody arms. In some embodiments, the antibody agent is a broadly neutralizing antibody agent (e.g., a broadly neutralizing antibody (bNAb)). A "broadly neutralizing antibody agent" is an antibody agent that has the ability to neutralize two or more genetic variants (e.g., strains) of a virus (e.g., HIV). In some embodiments, the antibody may lack covalent modifications (e.g., glycan linkages) that it would have if produced naturally. In some embodiments, antibodies may contain covalent modifications (e.g., attachment of glycans, payloads (e.g., detectable moieties, therapeutic moieties, catalytic moieties, etc.) or other pendant groups (e.g., polyethylene glycol, etc.)).In many embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence comprises one or more structural elements recognized by those skilled in the art as complementarity-determining regions (CDRs), and in some embodiments, an antibody agent is or comprises a polypeptide whose amino acid sequence comprises 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 that found in a reference antibody. In some embodiments, the included CDR is substantially identical to the reference CDR in that it either has an identical sequence or contains one to five amino acid substitutions compared to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that it exhibits at least 85%, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that it exhibits at least 95%, 96%, 97%, 98%, 99%, or 100% sequence identity with the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that at least one amino acid in the included CDR has been deleted, added, or substituted compared to the reference CDR, but the amino acid sequence of the included CDR is otherwise identical to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that 1 to 5 amino acids in the included CDR have been deleted, added, or substituted compared to the reference CDR, but the amino acid sequence of the included CDR is otherwise identical to the reference CDR. In some embodiments, the included CDR is substantially identical to the reference CDR in that at least one amino acid in the included CDR has been substituted compared to the reference CDR, but the amino acid sequence of the included CDR is otherwise identical to the reference CDR.In some embodiments, the included CDRs are substantially identical to the reference CDRs in that one to five amino acids in the included CDRs have been deleted, added, or substituted relative to the reference CDR, but the amino acid sequence of the included CDR is otherwise identical to the reference CDR. 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.

[0277] Aryl: The term "aryl" refers to monocyclic and bicyclic ring systems having a total of 6 to 14 ring members (e.g., C6-C14), wherein at least one ring in the system is aromatic and each ring in the system contains 3 to 7 ring members. In some embodiments, an "aryl" group contains a total of 6 to 12 ring members (e.g., C6-C12). The term "aryl" may be used interchangeably with the term "aryl ring." In certain embodiments, "aryl" refers to an aromatic ring system, including, but not limited to, phenyl, biphenyl, naphthyl, anthracyl, and the like, which may bear one or more substituents. Unless otherwise specified, an "aryl" group 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). Examples of fused aryl rings include: [ka] Examples include:

[0278] Associated: As used herein, two events or entities are "associated" with one another when the presence, level, degree, type, and / or form of one correlates with that of the other. For example, a particular entity (e.g., a polypeptide, genetic signature, metabolite, microorganism, etc.) is considered to be associated with a particular disease, disorder, or condition when its presence, level, and / or form correlates (e.g., across a relevant population) with the onset, susceptibility, severity, stage, etc. of the disease, disorder, or condition. In some embodiments, two or more entities are physically "associated" with one another when they directly or indirectly interact to bring them into and / or maintain them in physical proximity to one another. In some embodiments, two or more entities that are physically associated with one another are covalently linked to one another; in some embodiments, two or more entities that are physically associated with one another are not covalently linked to one another but are non-covalently associated, for example, by hydrogen bonding, van der Waals interactions, hydrophobic interactions, magnetism, and combinations thereof.

[0279] Co-administration: As used herein, the term "co-administration" refers to the use of a composition described herein (e.g., a pharmaceutical composition) with one or more additional therapeutic agents. In some embodiments, the one or more additional therapeutic agents comprise at least one polyribonucleotide encoding another antibody agent (e.g., an anti-HIV antigen antibody agent). The combination of a composition described herein (e.g., a pharmaceutical composition) with an additional therapeutic agent can be performed concurrently or separately (e.g., sequentially in any order). In some embodiments, a composition described herein (e.g., a pharmaceutical composition) and an additional therapeutic agent can be included together in a single pharmaceutically acceptable excipient, or they can be in separate excipients and delivered to a target cell or administered to a subject at different times. Each of these situations is contemplated as being within the meaning of "co-administration" or "combined use," provided that the composition described herein (e.g., a pharmaceutical composition) and the additional therapeutic agent are delivered or administered sufficiently closely in time that there is at least some temporal overlap in the biological effect(s) produced by each on the target cell or the subject being treated.

[0280] Combination therapy: As used herein, the term "combination therapy" refers to a situation in which a subject is exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents (e.g., two or more antibody agents)) simultaneously. In some embodiments, the two or more regimens may be administered simultaneously; in some embodiments, such regimens may be administered sequentially (e.g., all "doses" of a first regimen are administered prior to the administration of any doses of a second regimen); and in some embodiments, such agents are administered in overlapping dosing regimens. In some embodiments, administering combination therapy may involve administering one or more agent(s) or modality(s) to a subject receiving other agent(s) or modality(s) being administered in combination. For clarity, combination therapy does not require that the individual agents be administered together in a single composition (or, in some cases, necessarily simultaneously), although in some embodiments, two or more agents, or active portions thereof, may be administered together in a mixed composition. In some embodiments, the combination therapy includes polyribonucleotides encoding two or more antibody agents (eg, anti-HIV antibody agents).

[0281] Equivalent: As used herein, the term "equivalent" means that two or more agents, entities, circumstances, sets of conditions, etc., while not identical to one another, are sufficiently similar to permit them to be considered equivalent, such that one of skill in the art would recognize that reasonable conclusions can be drawn based on the observed differences or similarities. In some embodiments, equivalent sets of conditions, circumstances, individuals, or populations are characterized by multiple substantially identical characteristics and one or a few different characteristics. One of skill in the art will understand from the context what level of identity is required under any given circumstances for two or more such agents, entities, circumstances, sets of conditions, etc. to be considered equivalent. For example, one of skill in the art will understand that sets of circumstances, individuals, or populations are equivalent to one another when they are characterized by a sufficient number and type of substantially identical characteristics to warrant a reasonable conclusion that differences in results obtained or phenomena observed under or with different sets of circumstances, individuals, or populations are due to or indicate changes in the different characteristics.

[0282] Corresponding to: As used herein, the term "corresponding to" refers to a relationship between two or more entities. For example, the term "corresponding to" may be used to describe the position / identity of a structural element in a compound or composition as compared to another compound or composition (e.g., 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) may be identified as "corresponding to" a residue in an appropriate reference polymer. For example, those of skill in the art will recognize that, for simplicity, residues in a polypeptide are often designated using a canonical numbering system with reference to a reference related polypeptide, such that an amino acid "corresponding to," for example, a residue at position 190 corresponds to the residue found at 190 in the reference polypeptide, but not necessarily the actual 190th amino acid in a particular amino acid chain, and those of skill in the art will readily recognize how to identify a "corresponding" amino acid. For example, those of skill in the art will be aware of various sequence alignment strategies, e.g., software programs such as BLAST, CS-BLAST, CUSASW++, DIAMOND, FASTA, GGSEARCH / GLSEARCH, Genoogle, HMMER, HHpred / HHsearch, IDF, Infernal, KLAST, USARCH, parasail, PSI-BLAST, PSI-Search, ScalaBLAST, Sequilab, SAM, SSEARCH, SWAPHI, SWAPHI-LS, SWIMM, or SWIPE, that can be utilized in accordance with the present disclosure to identify "corresponding" residues in, for example, polypeptides and / or nucleic acids. Those of skill in the art will also recognize that the term "corresponding to" can in some cases be used to describe an event or entity that bears a meaningful similarity to another event or entity (e.g., an appropriate reference event or entity).By way of example only, a gene or protein of one organism may, in some embodiments, be said to "correspond to" a gene or protein from another organism to indicate that it plays a similar role or performs a similar function, and / or that it exhibits a particular degree of sequence identity or homology or has certain characteristic sequence elements in common.

[0283] Alicyclic: As used herein, "alicyclic" refers to a monocyclic C ring that has a single point of attachment or more than one point of attachment to the rest of the molecule, is fully saturated, or contains one or more units of unsaturation, but is not aromatic. 3-8 Hydrocarbon or bicyclic C 6-10 Refers to hydrocarbons.

[0284] Cycloalkyl: As used herein, the term "cycloalkyl" refers to an optionally substituted monocyclic or multicyclic saturated ring system of about 3 to about 10 carbon atoms. Exemplary monocyclic cycloalkyl rings include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and cycloheptyl.

[0285] Derived: With respect to an amino acid sequence (peptide or polypeptide) "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, an amino acid sequence derived from a particular amino acid sequence has an amino acid sequence that is identical, essentially identical, or homologous to the particular sequence or a fragment thereof. An amino acid sequence derived from a particular amino acid sequence may be a variant of the particular sequence or a fragment thereof. For example, an antibody agent utilized 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.

[0286] Detecting: The term "detecting" is used broadly herein to include any suitable means for determining the presence or absence of an entity of interest, or any form of measurement of an entity of interest in a sample. Thus, "detecting" can include determining, measuring, assessing, or testing the presence, absence, level, amount, and / or location of an entity of interest. Quantitative and qualitative determinations, measurements, or assessments are included, including semi-quantitative ones. Such determinations, measurements, or assessments can be relative, for example, when detecting an entity of interest relative to a control reference, or can be absolute. Thus, the term "quantifying" when used in the context of quantifying an entity of interest can refer to absolute or relative quantification. Absolute quantification can be achieved by correlating the detected level of the entity of interest with a known control standard (e.g., by constructing a standard curve). Alternatively, relative quantification can be achieved by comparing the detected levels or amounts 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.

[0287] Dosing regimen: Those skilled in the art will appreciate that the term "dosing regimen" (or "therapeutic regimen") can be used to refer to a set of unit doses (typically more than one) administered separately to a subject, typically spaced apart. In some embodiments, a given therapeutic agent has a recommended dosing regimen that can include one or more doses.

[0288] Encode: As used herein, the term "encode" or "code" refers to the sequence information of a first molecule that guides the production of a second molecule having a defined sequence of nucleotides (e.g., polyribonucleotides) or a defined sequence of amino acids. For example, a DNA molecule can encode an RNA molecule (e.g., by a transcription process involving a DNA-dependent RNA polymerase enzyme). An RNA molecule can encode a polypeptide (e.g., by a translation process). Thus, a gene, cDNA, or RNA molecule encodes a polypeptide when the polypeptide is produced in a cell or other biological system by transcription and translation of the RNA corresponding to that gene. In some embodiments, a coding region of a polyribonucleotide that encodes a target antigen refers to the coding strand, which has a nucleotide sequence identical to the polyribonucleotide sequence of such target antigen. In some embodiments, a coding region of a polyribonucleotide that encodes a target antigen refers to the non-coding strand of such target antigen, which can be used as a template for transcription of a gene or cDNA.

[0289] Engineered: In general, the term "engineered" refers to aspects that have been manipulated by human beings. For example, a polynucleotide is considered "engineered" when two or more sequences that are not naturally linked together in that order are manipulated by human beings to be directly linked to each other in the engineered polynucleotide, and / or when particular residues in the polynucleotide are linked by human acts to entities or moieties that do not occur in nature and / or are not naturally linked.

[0290] Epitope: As used herein, the term "epitope" refers to a portion that is specifically recognized by a binding component of an immunoglobulin (e.g., an antibody or receptor). For example, an epitope can be recognized by a T cell, a B cell, or an antibody. In some embodiments, an epitope is composed of multiple chemical atoms or groups on an antigen. In some embodiments, such chemical atoms or groups are surface-exposed when the antigen adopts a suitable conformation. In some embodiments, such chemical atoms or groups are physically near each other in space when the antigen adopts such a conformation. In some embodiments, at least some such chemical atoms or groups are physically separated from each other when the antigen adopts another conformation (e.g., linearized). Thus, in some embodiments, an epitope of an antigen can comprise 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 can 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.

[0291] 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 transcription product, e.g., 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.

[0292] Heteroaliphatic: The term "heteroaliphatic" or "heteroaliphatic group," as used herein, means an optionally substituted hydrocarbon moiety having, in addition to carbon atoms, 1 to 5 heteroatoms, which may be linear (i.e., unbranched), branched, or cyclic ("heterocyclic"), and which may be fully saturated or contain one or more units of unsaturation, 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 a basic nitrogen. The term "nitrogen" also includes substituted nitrogen. Unless otherwise specified, heteroaliphatic groups contain 1 to 10 carbon atoms, of which 1 to 3 carbon atoms are optionally and independently replaced by heteroatoms selected from oxygen, nitrogen, and sulfur. In some embodiments, heteroaliphatic groups contain 1 to 4 carbon atoms, of which 1 to 2 carbon atoms are optionally and independently replaced by heteroatoms selected from oxygen, nitrogen, and sulfur. In yet other embodiments, heteroaliphatic groups contain 1-3 carbon atoms, one of which is optionally and independently replaced by a heteroatom selected from oxygen, nitrogen, and sulfur. Suitable heteroaliphatic groups include, but are not limited to, linear or branched heteroalkyl, heteroalkenyl, and heteroalkynyl groups. For example, heteroaliphatic groups containing 1-10 atoms include the following exemplary groups: -O-CH, -CH-O-CH, -O-CH-CH-O-CH-CH-O-CH, etc.

[0293] Heteroaryl: The terms "heteroaryl" and "heteroara-," used alone or as part of a larger moiety, such as "heteroaralkyl" or "heteroaralkoxy," refer to a monocyclic or bicyclic ring group (e.g., a 5- or 6-membered monocyclic heteroaryl or a 9- or 10-membered bicyclic heteroaryl) having 5 to 10 ring atoms and having 6, 10, or 14 pi electrons shared in the cyclic arrangement and having 1 to 5 heteroatoms in addition to the carbon atoms. Heteroaryl groups include, but are not limited to, thienyl, furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl, oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl, thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl, purinyl, naphthyridinyl, pteridinyl, imidazo[1,2-a]pyrimidinyl, imidazo[1,2-a]pyridyl, imidazo[4,5-b]pyridyl, imidazo[4,5-c]pyridyl, pyrrolopyridyl, pyrrolopyrazinyl, thienopyrimidinyl, triazolopyridyl, and benzisoxazolyl. The terms "heteroaryl" and "heteroara-," as used herein, also include groups in which a heteroaromatic ring is fused to one or more aryl, alicyclic, or heterocyclyl rings, and the radical or point of attachment is on the heteroaromatic ring (i.e., a bicyclic heteroaryl ring having 1 to 3 heteroatoms). Non-limiting examples include indolyl, isoindolyl, benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl, benzotriazolyl, benzothiazolyl, benzothiadiazolyl, benzoxazolyl, quinolyl, isoquinolyl, 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 "heteroaryl ring," "heteroaryl group," or "heteroaromatic," all of which terms include rings that are optionally substituted.

[0294] 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.

[0295] 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 either saturated or partially unsaturated and has, in addition to carbon atoms, one or more, e.g., 1 to 4, heteroatoms as defined above. The term "nitrogen," when used in reference to a ring atom of a heterocycle, includes substituted nitrogen. By way of example, in a saturated or partially unsaturated ring having 0 to 3 heteroatoms selected from oxygen, sulfur, or nitrogen, the nitrogen can be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl), or NR (as in N-substituted pyrrolidinyl). +Heterocyclic rings can be attached to their pendant groups at any heteroatom or carbon atom that results in a stable structure, and any of the ring atoms can be optionally substituted. Examples of such saturated or partially unsaturated heterocyclic radicals include, but are not limited to, azetidinyl, oxetanyl, tetrahydrofuranyl, tetrahydrothienyl, pyrrolidinyl, piperidinyl, decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and thiamorpholinyl. Heterocyclyl groups can be monocyclic, bicyclic, tricyclic, or polycyclic, preferably monocyclic, bicyclic, or tricyclic, and more preferably monocyclic or bicyclic. Bicyclic heterocyclic rings also include groups in which a heterocyclic ring is fused with one or more aryl rings. Exemplary bicyclic heterocyclic groups include indolinyl, isoindolinyl, benzodioxolyl, 1,3-dihydroisobenzofuranyl, 2,3-dihydrobenzofuranyl, and tetrahydroquinolinyl. The bicyclic heterocyclic ring may also be a spirocyclic ring system (e.g., a 7- to 11-membered spirocyclic fused heterocyclic ring having, in addition to carbon atoms, one or more heteroatoms (e.g., 1, 2, 3, or 4 heteroatoms) as defined above). The bicyclic heterocyclic ring may also be a bridged ring system (e.g., a 7- to 11-membered bridged heterocyclic ring having 1, 2, or 3 bridging atoms).

[0296] Homology: As used herein, the terms "homology" or "homologues" refer to the overall relatedness between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or 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 one another 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, polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or polypeptide molecules are considered to be "homologous" to one another if their sequences are at least 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% similar (e.g., contain residues with related chemical properties at corresponding positions). For example, as is well known to those of skill in the art, certain amino acids are typically classified as being similar to one another as being "hydrophobic" or "hydrophilic" amino acids and / or as having "polar" or "nonpolar" side chains. Substitution of one amino acid for another of the same type can often be considered a "homologous" substitution.

[0297] 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 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 one another if their sequences are at least 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99% identical. Calculation of the percent identity of two nucleic acid or polypeptide sequences can be performed, for example, by aligning the two sequences for 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 disregarded for comparison purposes). In certain embodiments, the length of the sequences aligned for comparison purposes 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. Nucleotides at corresponding positions are then compared. If a 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 that position. The percent identity between two sequences is a function of the number of identical positions shared between the sequences, taking into account the number of gaps and the length of each gap that need to be introduced to optimally align the two sequences. Sequence comparison and determination of the percent identity between two sequences can be accomplished using a mathematical algorithm. For example, percent identity between two nucleotide sequences can be determined using the algorithm of Meyers and Miller (1989), which is incorporated into the ALIGN program (version 2.0).In some exemplary embodiments, nucleic acid sequence comparisons performed using the ALIGN program use a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Alternatively, the percent identity between two nucleotide sequences can be determined using the GAP program included in the GCG software package using the NWSgapdna.CMP matrix.

[0298] Increased, induced, or reduced: As used herein, these terms, or grammatically equivalent comparative terms, refer to a value relative to an equivalent reference measurement. For example, in some embodiments, an assessment value obtained using a provided composition (e.g., a pharmaceutical composition) may be "increased" compared to an assessment value obtained using an equivalent reference composition. Alternatively, or in addition, in some embodiments, an assessment value obtained in a subject may be "increased" compared to an assessment value obtained in the same subject under different conditions (e.g., before or after an event, or in the presence or absence of an event such as administration of a composition (e.g., a pharmaceutical composition) described herein), or in a different, comparable subject (e.g., a comparable subject that differs from the subject of interest in that it has previously been exposed to a condition, e.g., in the absence of administration of a composition (e.g., a pharmaceutical composition) described herein). In some embodiments, comparative terms refer to a statistically significant difference (e.g., having sufficient dominance and / or magnitude to achieve statistical significance). Those skilled in the art will know, or can readily determine, the degree and / or magnitude of difference required or sufficient to achieve such statistical significance in a given context. In some embodiments, the term "reduced" or equivalent terms refers to a reduction in the level of an evaluated value by at least 5%, at least 10%, at least 20%, at least 50%, at least 75%, or more relative to a comparable reference. In some embodiments, the term "reduced" or equivalent terms refers to complete or substantially complete inhibition, i.e., a reduction to zero or substantially zero. In some embodiments, the term "increased" or "induced" refers to a reduction in the level of an evaluated 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 relative to an equivalent reference.

[0299] In order: As used herein with respect to a polynucleotide or polyribonucleotide, "in order" refers to the order of features along the polynucleotide or polyribonucleotide from 5' to 3'. As used herein with respect to a polypeptide, "in order" refers to the order of features along the polypeptide, moving from the most N-terminal feature to the most C-terminal feature. "In order" does not mean that there cannot be additional features between the listed features. For example, if features A, B, and C of a polynucleotide are described herein as being "in order: feature A, feature B, and feature C," this description does not exclude, for example, feature D being located between feature A and feature B.

[0300] Ionizable: The term "ionizable" refers to a compound or group or atom that is charged at a particular pH. In the context of an ionizable amino lipid, such lipid or its functional group or atom is positively charged at a particular pH. In some embodiments, the ionizable amino lipid is positively charged at an acidic pH. In some embodiments, the ionizable amino lipid is primarily neutral at physiological pH values, e.g., in some embodiments, about 7.0 to 7.4, but becomes positively charged at lower pH values. In some embodiments, the ionizable amino lipid may have a pKa in the range of about 5 to about 7.

[0301] Isolated: The term "isolated" means changed or removed from the natural state. For example, a nucleic acid or peptide that is naturally present in a living animal is not "isolated," but the same nucleic acid or peptide partially or completely separated from the coexisting materials in its natural state is "isolated." An isolated nucleic acid or protein can exist in a substantially purified form, or can exist in a non-native environment, such as, for example, a host cell.

[0302] Lipid: As used herein, the terms "lipid" and "lipid-like substance" are broadly defined as molecules that contain one or more hydrophobic moieties or groups and, optionally, one or more hydrophilic moieties or groups. Molecules that contain hydrophobic and hydrophilic moieties are also typically referred to as amphiphiles.

[0303] RNA lipid nanoparticles: As used herein, the term "RNA lipid nanoparticles" refers to nanoparticles comprising at least one lipid and RNA molecule(s), such as one or more polyribonucleotides 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 described herein may have an average particle 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 may have a particle size (e.g., Z-average) 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 nm to about 90 nm, about 80 nm to about 90 nm, or about 70 nm to about 80 nm. In some embodiments, the average particle size of the lipid nanoparticles is determined by measuring the average particle diameter. In some embodiments, the RNA-lipid nanoparticles may be prepared by mixing lipids with the RNA molecules described herein.

[0304] 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, e.g., a receptor-binding protein, thereby inhibiting parasitic infection of a cell. In some embodiments, the term "neutralization" refers to an event in which the ability to infect a cell is eliminated or significantly reduced by the binding agent.

[0305] Nucleic Acid / Polynucleotide: As used herein, the term "nucleic acid" refers to a polymer of at least 10 or more nucleotides. In some embodiments, a nucleic acid is or comprises DNA. In some embodiments, a nucleic acid is or comprises RNA. In some embodiments, a nucleic acid is or comprises peptide nucleic acid (PNA). In some embodiments, a nucleic acid is or comprises single-stranded nucleic acid. In some embodiments, a nucleic acid is or comprises double-stranded nucleic acid. In some embodiments, a nucleic acid comprises both single-stranded and double-stranded portions. In some embodiments, a nucleic acid comprises a backbone comprising one or more phosphodiester bonds. In some embodiments, a nucleic acid comprises a backbone comprising both phosphodiester and non-phosphodiester bonds. For example, in some embodiments, a nucleic acid can comprise a backbone with one or more phosphorothioate or 5'-N-phosphoramidite linkages, and / or one or more peptide bonds, e.g., as in "peptide nucleic acids." In some embodiments, a nucleic acid comprises one or more or all naturally occurring residues (e.g., adenine, cytosine, deoxyadenosine, deoxycytidine, deoxyguanosine, deoxythymidine, guanine, thymine, uracil). In some embodiments, a nucleic acid comprises one or more or all non-naturally occurring 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, intercalated 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) compared to that of a natural residue. In some embodiments, the nucleic acid has a nucleotide sequence that encodes a functional gene product, e.g., an RNA or a polypeptide. In some embodiments, the nucleic acid has a nucleotide sequence that includes one or more introns. In some embodiments, the nucleic acid can be prepared by isolation from a natural source, enzymatic synthesis (e.g., by polymerization based on a complementary template, e.g., 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, 110, 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, 8000, 9000, 1000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1800, 1900, 2000, 2250, 2500, 275, 3000, 3250, 3500, 375, 4000, 4250, 4500, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, and 500, 5000, 5500, 6000, 6500, 7000, 7500, 8000, 8500, 9000, 9500, 10,000, 10,500, 11,000, 11,500, 12,000, 12,500, 13,000, 13,500, 14,000, 14,500, 15,000, 15,500, 16,000, 16,500, 17,000, 17,500, 18,000, 18,500, 19,000, 19,500, or 20,000 or more residues or nucleotides in length.

[0306] Pharmaceutically effective amount: The term "pharmaceutically effective amount" or "therapeutically effective amount" refers to an amount that alone or together with further doses produces a desired response or a desired effect. In the case of treating a particular disease (e.g., HIV), the desired response, in some embodiments, relates to inhibiting the course of the disease (e.g., HIV). In some embodiments, such inhibition can include slowing the progression of the disease (e.g., HIV) and / or arresting or reversing the progression of the disease (e.g., HIV). In some embodiments, the desired response in treating a disease (e.g., HIV) can be or can include delaying or preventing the onset of the disease (e.g., HIV) or condition (e.g., an HIV-associated condition). An effective amount of a composition (e.g., a pharmaceutical composition) described herein will depend, for example, on the disease (e.g., HIV) or condition (e.g., an HIV-associated condition) being treated, the severity of such disease (e.g., HIV) or condition (e.g., an HIV-associated condition), the patient's personal parameters, such as age, physiological condition, size, and weight, the duration of treatment, the type of concomitant therapy (if any), the particular route of administration, and similar factors. Thus, the dose of a composition (e.g., a pharmaceutical composition) described herein may depend on various such parameters. If the patient does not respond adequately to the initial dose, a higher dose (or an effectively higher dose achieved by an alternative, more localized route of administration) may be used.

[0307] Polypeptide: As used herein, the term "polypeptide" refers to a polymeric chain of amino acids. In some embodiments, a polypeptide has a naturally occurring amino acid sequence. In some embodiments, a polypeptide has a non-naturally occurring amino acid sequence. In some embodiments, a polypeptide has an amino acid sequence that is engineered, in that it has been designed and / or produced by human activity. In some embodiments, a polypeptide can comprise or consist of natural amino acids, unnatural amino acids, or both. In some embodiments, a polypeptide can comprise or consist of only natural amino acids or only unnatural amino acids. In some embodiments, a polypeptide can comprise D-amino acids, L-amino acids, or both. In some embodiments, a polypeptide can comprise only D-amino acids. In some embodiments, a polypeptide can comprise only L-amino acids. In some embodiments, a polypeptide can comprise one or more pendant groups or other modifications, e.g., modifying or attached to one or more amino acid side chains, at the N-terminus of the polypeptide, the C-terminus of the polypeptide, or any combination thereof. In some embodiments, such pendant groups or modifications include acetylation, amidation, lipidation, methylation, pegylation, etc. (including combinations thereof). In some embodiments, a polypeptide may be cyclic and / or include a cyclic portion. In some embodiments, a polypeptide is not cyclic and / or does not include a cyclic portion. In some embodiments, a polypeptide is linear. In some embodiments, a polypeptide may be or include a stapled polypeptide. In some embodiments, the term "polypeptide" may be attached to the name of a reference polypeptide, activity, or structure, and in such cases, it is used to refer to polypeptides that share a related activity or structure and therefore may be considered members of the same class or family of polypeptides. For each such class, exemplary polypeptides within the class whose amino acid sequence and / or function are known are provided herein and / or will be known to those of skill in the art.In some embodiments, such exemplary polypeptides are reference polypeptides of a class or family of polypeptides. In some embodiments, members of a class or family of polypeptides exhibit significant sequence homology or identity with, share common sequence motifs (e.g., characteristic sequence elements) with, and / or share common activity (in some embodiments, at a similar level or within a specified range) with the reference polypeptide of the class (in some embodiments, all polypeptides within the class). For example, in some embodiments, a member polypeptide exhibits a degree of overall sequence homology or identity with a reference polypeptide of at least about 30-40%, often greater than about 50%, 60%, 70%, 80%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or more, and / or comprises at least one region (e.g., a conserved region, which in some embodiments may be or may include a distinctive sequence element) that exhibits very high, often greater than 90%, or even greater than 95%, 96%, 97%, 98%, or 99% sequence identity. Such conserved regions typically encompass at least 3-4 amino acids, and often up to 35 or more, and in some embodiments, the 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 consecutive amino acids. In some embodiments, the related polypeptide may comprise or consist of a fragment of a parent polypeptide.

[0308] Prevent: As used herein, the term "prevent" or "prevention," when used in reference to the occurrence of a disease, disorder, and / or condition, means reducing the risk of developing the disease, disorder, and / or condition and / or delaying the onset of one or more features or symptoms of the disease, disorder, or condition. Prevention may be considered complete when the onset of the disease, disorder, or condition has been delayed by a predetermined period of time.

[0309] Reference: As used herein, the term "reference" refers to a standard or control against 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, the reference or control is tested and / or measured substantially simultaneously with the testing or measurement of the agent of interest. In some embodiments, the reference or control is a historical reference or control, optionally incorporated into a tangible medium. As will be understood by those of skill in the art, a reference or control is typically measured or characterized under conditions or circumstances comparable to those being evaluated. Those of skill in the art will be familiar with when sufficient similarity exists to justify reliance on and / or equivalence to a particular reference or control being considered.

[0310] Ribonucleic Acid (RNA) or Polyribonucleotide: As used herein, the terms "ribonucleic acid," "RNA," or "polyribonucleotide" refer to a polymer of ribonucleotides. In some embodiments, the RNA is single-stranded. In some embodiments, the RNA is double-stranded. In some embodiments, the RNA contains both single-stranded and double-stranded portions. In some embodiments, the RNA may contain a backbone structure described in the definition of "nucleic acid / polynucleotide" above. The RNA may be a regulatory RNA (e.g., siRNA, microRNA, etc.) or a messenger RNA (mRNA). In some embodiments, the RNA is mRNA. In some embodiments, the RNA is mRNA, the RNA typically contains a poly(A) region at its 3' end. In some embodiments, the RNA is mRNA, the RNA typically contains an art-recognized cap structure at its 5' end for ribosome recognition and binding of the mRNA to initiate translation. In some embodiments, the RNA is synthetic RNA. Synthetic RNA includes RNA synthesized in vitro (e.g., by enzymatic synthesis and / or chemical synthesis).

[0311] Ribonucleotide: As used herein, the term "ribonucleotide" encompasses unmodified ribonucleotides and modified ribonucleotides. For example, unmodified ribonucleotides include the purine bases adenine (A) and guanine (G) and the pyrimidine bases cytosine (C) and uracil (U). Modified ribonucleotides can include one or more modifications, including, but not limited to, (a) terminal modifications, such as 5'-terminal modifications (e.g., phosphorylation, dephosphorylation, conjugation, reverse linkage, etc.), 3'-terminal modifications (e.g., conjugation, reverse linkage, etc.), (b) base modifications, such as replacement with a modified base, a stabilizing base, a destabilizing base, or a base that base-pairs with an expanded repertoire partner, or a conjugated base), (c) sugar modifications or sugar replacements (e.g., at the 2' or 4' position), and (d) internucleoside linkage modifications, such as modification or replacement of a phosphodiester bond. The term "ribonucleotide" also encompasses ribonucleotide triphosphates, including modified and unmodified ribonucleotide triphosphates.

[0312] Risk: As will be understood from the context, "risk" of a disease, disorder, and / or condition refers to the likelihood that a particular individual will develop the disease, disorder, and / or condition. In some embodiments, risk is expressed as a percentage. In some embodiments, risk is expressed as risk relative to the risk associated with a reference sample or group of reference samples. In some embodiments, the reference sample or group of reference samples has a known risk of the disease, disorder, condition, and / or event. In some embodiments, the reference sample or group of reference samples is from an individual that is comparable to the particular individual. In some embodiments, risk may reflect one or more genetic attributes that may, for example, predispose an individual to developing (or not developing) a particular disease, disorder, and / or condition. In some embodiments, risk may reflect one or more epigenetic events or attributes, and / or one or more lifestyle or environmental events or attributes.

[0313] Selective or specific: The terms "selective" or "specific," as used herein with respect to an active agent, will be understood by those skilled in the art to mean that the agent discriminates among potential target entities, conditions, or cells. For example, in some embodiments, an agent is said to bind "specifically" to its target if it preferentially binds to that target in the presence of one or more competing targets. In many embodiments, the specific interaction is dependent on the presence of a particular structural feature of the targeting entity (e.g., an epitope, cleft, binding site). It should be understood that specificity need not be absolute. In some embodiments, specificity can be assessed relative to the specificity of the target-binding moiety for one or more other potential target entities (e.g., competitors). In some embodiments, specificity is assessed relative to the specificity of a specific-binding reference moiety. In some embodiments, specificity is assessed relative to the specificity of a non-specific-binding reference moiety.

[0314] Substituted or Optionally Substituted: As described herein, compounds of the invention may contain "optionally substituted" moieties. Whether preceded by the term "optionally" or not, the term "substituted" generally means that one or more hydrogens of the specified moiety are replaced with a suitable substituent. "Substituted" applies to one or more hydrogens either stated or implied by the structure (e.g., [ka] At least [ka] refers to, [ka] At least [ka] (refers to "optionally substituted"). Unless otherwise specified, an "optionally substituted" group may have a suitable substituent at each substitutable position of the group, and if more than one position in any given structure may be substituted with more than one substituent selected from a specified group, the substituents at each position may be either the same or different. Combinations of substituents envisioned by the present invention are preferably those that result in the formation of stable or chemically feasible compounds. The term "stable," as used herein, refers to a compound that remains substantially unchanged when exposed to conditions that permit its production, detection, and, in certain embodiments, its recovery, purification, and use for one or more of the purposes provided herein. Groups described as "substituted" preferably have 1 to 4 substituents, more preferably 1 or 2 substituents. Groups described as "optionally substituted" may be unsubstituted or "substituted" as described above.

[0315] Suitable monovalent substituents on a substitutable carbon atom of an "optionally substituted" group are independently halogen; -(CH) 0-4 R°;-(CH2) 0-4 OR°;-O(CH2) 0-4 R o ,-O-(CH2) 0-4 C(O)OR°;-(CH2) 0-4 CH(OR°)2;-(CH2) 0-4 SR°; optionally substituted with R° -(CH2) 0-4 Ph; optionally substituted with R° -(CH2) 0-4 O(CH2) 0-1 Ph; optionally substituted with R° -CH=CHPh; optionally substituted with R° -(CH2) 0-4 O(CH2) 0-1 -pyridyl; -NO2; -CN; -N3; ​​-(CH2) 0-4 N(R°)2;-(CH2) 0-4 N(R°)C(O)R°;-N(R°)C(S)R°;-(CH2) 0-4 N(R°)C(O)NR°2;-N(R°)C(S)NR°2;-(CH2)0-4 N(R°)C(O)OR°;-N(R°)N(R°)C(O)R°;-N(R°)N(R°)C(O)NR°2;-N(R°)N(R°)C(O)OR°;-(CH2) 0-4 C(O)R°;C(S)R°;-(CH2) 0-4 C(O)OR°;-(CH2) 0-4 C(O)SR°;-(CH2) 0-4 C(O)OSiR°3;-(CH2) 0-4 OC(O)R°;-OC(O)(CH2) 0-4 SR°;-(CH2) 0-4 SC(O)R°;-(CH2) 0-4 C(O)NR°2;-C(S)NR°2;-C(S)SR°;-SC(S)SR°,-(CH2) 0-4 OC(O)NR°2;-C(O)N(OR°)R°;-C(O)C(O)R°;-C(O)CH2C(O)R°;-C(NOR°)R°;-(CH2) 0-4 SSR°;-(CH2) 0-4 S(O)2R°;-(CH2) 0-4 S(O)2OR°;-(CH2) 0-4 OS(O)2R°;-S(O)2NR°2;-(CH2) 0-4 S(O)R°;-N(R°)S(O)2NR°2;-N(R°)S(O)2R°;-N(OR°)R°;-C(NH)NR°2;-P(O)2R°;-P(O)R°2;-OP(O)R°2;-OP(O)(OR°)2;SiR°3;-(C 1-4 Linear or branched alkylene)ON(R°)2; or -(C 1-4 linear or branched alkylene)C(O)ON(R°)2, where each R° may be optionally substituted as defined below and independently represents hydrogen, C 1-6 Aliphatic, -CH2Ph, -O(CH2) 0-1Ph, -CH2- (a 5- or 6-membered heteroaryl ring), or a 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or, notwithstanding the above, two independently occurring R° together with the intervening atom(s) form a 3- to 12-membered saturated, partially unsaturated, or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur, which may be substituted as defined below.

[0316] Suitable monovalent substituents on R° (or the ring formed by two independent occurrences of R° taken together with the atom(s) intervening therebetween) are independently halogen, —(CH) 0-2 R ● ,-(Halo R ● ), -(CH2) 0-2 OH, -(CH2) 0-2 OR ● , -(CH2) 0-2 CH(OR ● )2, -O(HaloR ●· ), -CN, -N3, -(CH2) 0-2 C(O)R ● , -(CH2) 0-2 C(O)OH, -(CH2) 0-2 C(O)OR ● , -(CH2) 0-2 SR ● , -(CH2) 0-2 SH, -(CH2) 0-2 NH2, -(CH2) 0-2 NHR ● , -(CH2) 0-2 NR ● 2, -NO2, -SiR ● 3. -OSiR ● 3. -C(O)SR ● 、 -(C 1-4 Linear or branched alkylene)C(O)OR ● , or -SSR ● where each R ●is unsubstituted or, if preceded by "halo", substituted only with one or more halogens, and independently, C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 or a 3-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Suitable divalent substituents on a saturated carbon atom of R° include ═O and ═S.

[0317] Suitable divalent substituents on a saturated carbon atom of an "optionally substituted" group include the following: ═O ("oxo"), ═S, ═NNR * 2, =NNHC(O)R * , =NNHC(O)OR * , =NNHS(O)2R * , =NR * , =NOR * , -O(C(R * 2)) 2-3 O-, or -S(C(R * 2)) 2-3 S-, and each R occurring independently in the formula * is hydrogen, optionally substituted as defined below 1-6 A 5- to 6-membered, saturated, partially unsaturated, or aryl ring is an aliphatic or unsubstituted group having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur. Preferred divalent substituents attached to vicinal substitutable carbon atoms of an "optionally substituted" group include -O(CR * 2) 2-3 O-, and each R occurring independently in the formula * is hydrogen, optionally substituted as defined below 1-6 It is selected from aliphatic or unsubstituted 5-6 membered saturated, partially unsaturated or aryl rings having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0318] R * Suitable substituents on the aliphatic group include halogen, -R ● ,-(Halo R ● ), -OH, -OR● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● , -NR ● 2, or -NO2, wherein each R ● is unsubstituted or, if preceded by "halo", substituted only with one or more halogens, and independently, C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 3-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0319] Suitable substituents on a substitutable nitrogen of an "optionally substituted" group include -R † , -NR † 2. -C(O)R † , -C(O)OR † , -C(O)C(O)R † , -C(O)CHC(O)R † , -S(O)2R † , -S(O)NR † 2. -C(S)NR † 2. -C(NH)NR † 2, or -N(R † )S(O)2R † wherein each R † are independently hydrogen, optionally substituted as defined below, C 1-6 an aliphatic, unsubstituted -OPh, or an unsubstituted 3- to 6-membered saturated, partially unsaturated, or aryl ring having 0 to 4 heteroatoms independently selected from nitrogen, oxygen, or sulfur; or, notwithstanding the above, two independently occurring R † together with the intervening atom(s) form an unsubstituted 3-12 membered saturated, partially unsaturated or aryl monocyclic or bicyclic ring having 0-4 heteroatoms independently selected from nitrogen, oxygen or sulfur.

[0320] R †Suitable substituents on the aliphatic group are independently halogen, —R ● ,-(Halo R ● ), -OH, -OR ● , -O(HaloR ● ), -CN, -C(O)OH, -C(O)OR ● , -NH2, -NHR ● ,-NR ● 2, or -NO2, where each R ● is unsubstituted or, if preceded by "halo", substituted only with one or more halogens, and independently, C 1-4 Aliphatic, -CH2Ph, -O(CH2) 0-1 Ph, or a 3-6 membered saturated, partially unsaturated, or aryl ring having 0-4 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

[0321] Subject: As used herein, the term "subject" refers to an organism to which a composition described herein is administered, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. Typical subjects include animals (e.g., mammals, e.g., 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 condition (e.g., HIV, an HIV-related condition, etc.). In some embodiments, the subject is predisposed to a disease, disorder, or condition (e.g., HIV, an HIV-related condition, etc.). In some embodiments, the subject exhibits one or more symptoms or characteristics of a disease, disorder, or condition (e.g., HIV, an HIV-related condition, etc.). In some embodiments, the subject exhibits one or more non-specific symptoms of a disease, disorder, or condition (e.g., HIV, an HIV-related condition, etc.). In some embodiments, the subject does not exhibit any symptoms or characteristics of a disease, disorder, or condition (e.g., HIV, an HIV-related condition, etc.). In some embodiments, the subject has one or more features characteristic of susceptibility to or risk for a disease, disorder, or condition (e.g., HIV, an HIV-related condition, etc.). In some embodiments, the subject is a patient. In some embodiments, the subject is an individual for whom and / or to whom a diagnosis and / or therapy is being administered.

[0322] Suffering from: An individual who is "suffering from" a disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.) has been diagnosed with and / or exhibits the disease, disorder, and / or condition.

[0323] Susceptible: An individual who is "susceptible" to a disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.) is an individual who is at a higher risk of developing the disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.) compared to the general population. In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.) may not have been diagnosed with the disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.) may exhibit symptoms of the disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.) may not exhibit symptoms of the disease, disorder, and / or condition (e.g., HIV, HIV-related conditions, etc.). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., HIV, an HIV-related condition, etc.) will develop the disease, disorder, and / or condition (e.g., HIV, an HIV-related condition, etc.). In some embodiments, an individual who is susceptible to a disease, disorder, and / or condition (e.g., HIV, an HIV-related condition, etc.) will not develop the disease, disorder, and / or condition (e.g., HIV, an HIV-related condition, etc.).

[0324] 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., demonstrated to have a statistically likely likelihood of having 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, reduce, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition (e.g., HIV, HIV-related illnesses, etc.). In some embodiments, a therapeutic agent or therapy is a medical intervention (e.g., surgery, radiation, phototherapy) that can be performed to alleviate, alleviate, inhibit, present, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition.

[0325] Treat: As used herein, the terms "treat," "treatment," or "treating" refer to any method that may be used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or characteristics of a disease, disorder, and / or condition (e.g., HIV, HIV-related illnesses, etc.). Treatment may be administered to a subject who does not exhibit symptoms of a disease, disorder, and / or condition (e.g., HIV, HIV-related illnesses, etc.). In some embodiments, treatment may be administered to a subject who exhibits only early signs of a disease, disorder, and / or condition (e.g., HIV, HIV-related illnesses, etc.), e.g., to reduce the risk of developing symptoms associated with the disease, disorder, and / or condition. In some embodiments, treatment may be administered to a subject at a later stage of a disease, disorder, and / or condition (e.g., HIV, HIV-related illnesses, etc.).

[0326] Detailed Description of Specific Embodiments I. Human immunodeficiency virus (HIV) Human immunodeficiency virus (HIV) is a lentivirus within the retroviridae family. Mature HIV particles are roughly circular and approximately 100 nm in diameter. They consist of a core composed of two identical single-stranded RNA molecules, a capsid, and an envelope (Figure 1, Panel B) (Musumeci et al., Molecules 20.9(2015):17511-17532, incorporated herein by reference). The envelope consists of a lipid bilayer and Env proteins. These Env proteins exist as trimers of the gp120 surface protein anchored to the envelope membrane via the gp41 transmembrane protein. The viral capsid, surrounded by the envelope, contains a symmetric outer capsid membrane composed of the matrix protein p17. Inside the outer capsid membrane is a conical capsid composed of the inner capsid protein p24. The inner capsid is attached to the outer capsid membrane at its tapered cone. The inner capsid contains viral RNA (two identical copies) and the viral enzymes reverse transcriptase, integrase, and protease. Also contained within the viral particle are oligopeptides generated by proteolytic processing of Gag and Gag / Pol precursor proteins p55 and p160 during viral particle maturation (GAC, Transfusion Medicine Hemotherapy, 43:203-222, 2016, incorporated herein by reference).

[0327] There are two main types of HIV: HIV-1 and HIV-2. HIV-1 is the most common type of HIV, accounting for 95% of all infections worldwide. HIV-2 is relatively rare and less contagious. HIV-2 is primarily concentrated in West Africa and surrounding countries.

[0328] HIV-1 and HIV-2 share many similarities, including their intracellular replication pathways, modes of transmission, and the clinical consequences that lead to acquired immune deficiency syndrome (AIDS). However, HIV-2 is less likely to progress to AIDS due to its lower transmissibility. Thus, individuals infected with HIV-2 generally do not experience long-term disease progression, while patients infected with HIV-1 tend to progress more rapidly to AIDS.

[0329] However, once progression begins, the pathological processes of both viruses are similar. One difference is that HIV-2 has been found to progress at higher CD4 counts. Furthermore, HIV-2 infection is characterized by a lower viral load of over 10,000 copies / mL compared to millions of copies / mL for HIV-1. In the case of HIV-2 infection, the subject's immune response tends to be more protective, thus slowing the progression of the disease.

[0330] HIV-1 and HIV-2 are in turn further classified into groups and subtypes. HIV-1 is classified into major or M group, aberrant or O group, and non-M / O or N group. The most common group is group M, which is the primary cause of the global HIV epidemic. Other groups are relatively rare and are found in limited geographies, including Gabon, Cameroon, and Equatorial Guinea.

[0331] Group M is further subdivided into genetically distinct subtypes: A, B, C, D, F, G, H, J, and K. Some of these subtypes combine to form hybrid viruses called "recombinant epidemic strains." Subtype B accounts for 12% of HIV infections worldwide. Subtype B is the predominant HIV-1 subtype found in the Americas, Australasia, and Western Europe. Consequently, most clinical research on HIV to date has focused on these populations.

[0332] Subtype C accounts for nearly 50% of all HIV-positive individuals, yet fewer studies have focused on this subtype. Subtype C is common in southern African countries, where HIV incidence is extremely high. A wide variety of HIV-1 subtypes exist in Cameroon and the Democratic Republic of the Congo, the HIV-1 epicenters. However, global subtype distribution patterns are currently changing due to population mixing and migration.

[0333] To date, approximately eight HIV-2 subtypes have been identified. The two major subtypes of HIV-2 that are considered endemic are A and B. HIV-2 group A infections are primarily found in West Africa, with a few cases reported in Brazil, Europe, the United States, and India. HIV-2 group B infections are found only in West Africa.

[0334] Because HIV subtypes can vary geographically, an ideal therapeutic agent would be one that can target and neutralize more than one subtype of HIV, and even more preferably multiple strains. As discussed further below, anti-HIV antibodies have been developed that have the ability to bind to and at least temporarily neutralize HIV virions. Nevertheless, problems with such anti-HIV antibodies remain, including challenges with administration, antibody persistence in vivo, and viral escape. The polyribonucleotides and compositions of the present disclosure, as described herein, address these challenges.

[0335] A. The HIV genome HIV contains two identical copies of single-stranded DNA encoding its genome. When the virus integrates into a host cell, reverse transcription of viral RNA into double-stranded DNA occurs, which leads to RNA degradation and integration of the double-stranded DNA or proviral DNA into the host genome. The HIV genome is flanked at both ends by LTR (long terminal repeat) regions, including the 5' LTR, which encodes a transcription promoter. The RNA genome is 9749 nucleotides long 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).

[0336] The HIV genome contains the following genes: gag, pol, vif, vpr, tat, rev, vpu, env, and nef (see Figure 1, panel A). The proteins encoded by gag, pol, and env are viral structural proteins. The proteins encoded by tat and ref are essential regulatory proteins. The proteins encoded by nef, vpr, vif, and vpu are auxiliary regulatory proteins. The gag gene encodes the P555Gag precursor protein, which is composed of the outer core membrane (p17), capsid protein (p24), nucleoprotein (p7), Pr55Gag, and p6 proteins. Protein p24 forms the cone-shaped capsid, and protein p17 forms the inner membrane layer. Protein p6 is involved in the release of virus particles.

[0337] The pol gene encodes the Pr160GagPol precursor protein, the protease enzymes p10, 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) yields the protease p10. The p51 reverse transcriptase protein is responsible for the transcription of HIV RNA into proviral DNA. The p55 (RNase H) protein functions to degrade viral RNA in the viral RNA / DNA complex to generate proviral DNA. The p32 integrase protein functions in integrating the proviral DNA into the host cell genome.

[0338] The env gene encodes the precursor protein PrGp160 of two envelope glycoproteins, gp120 (surface protein) and gp41 (transmembrane protein). Proteins gp120 and gp41 are generated by protease cleavage of the precursor protein PrGp160. Protein gp120 functions when the virus binds to target host cells. Protein gp41 functions to anchor gp120 to the viral membrane and fuse the viral membrane with the target cell membrane.

[0339] The tat gene encodes the Tat protein p14 (transactivator protein), which activates viral gene transcription. The rev gene encodes the Rev protein p19 (RNA splicing regulator), which regulates the export of mRNA (both unspliced ​​and partially spliced). The nef gene encodes the Nef protein p27 (negative regulator), which functions during HIV replication to enhance viral infectivity in host cells. The p27 protein also functions to downregulate CD4 and HLA on target cells. The vif gene encodes the Vif protein p23 (viral infectivity factor), which functions during viral production in host cells. The vpr gene encodes the Vpr protein p15 (viral protein r). This protein interacts with the p6 protein to promote viral infectivity in host cells. The gene vpu encodes the Vpu protein p16 (viral protein unique), which enables efficient release of viral particles and controls CD4 degradation on target cells. The protein p16 also controls intracellular signaling. The gene vpx encodes the Vpx protein p15 (viral protein x), which functions to interact 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).

[0340] B. Life cycle The HIV life cycle 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 contact target host cells. The surface glycoprotein env gp120 of mature HIV particles binds to the CD4 receptor on the target host cell, which initiates further binding of gp120 to co-receptors, i.e., chemokine receptor 5 (CCR5) or chemokine receptor 4 (CXCR4 for fusin). Binding of gp120 to CD4 and the co-receptor induces a conformational change in gp120 so that gp41 is displayed on the viral membrane and can fuse with the plasma membrane of the target host cell. The viral capsid then enters the host cell cytoplasm. The capsid is internalized by an endosome and releases its contents, i.e., viral RNA. Entry and release of the virus into the target host cell causes the virus to undergo reverse transcription, which reverse-transcribes viral RNA into single-stranded cDNA. The RNA strand is then degraded by RNase H, and the single-stranded cDNA is converted into double-stranded DNA by the DNA-dependent DNA polymerase activity of the enzyme reverse transcriptase.

[0341] Double-stranded DNA, or proviral DNA, forms a complex with integrase, is transported into the host cell nucleus, and randomly inserts itself into the host cell genome. Once integrated into the genome, the proviral genome is replicated. 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 binding sites for cellular DNA-dependent RNA polymerase and transcription factors to initiate transcription. Transcription of the proviral DNA is accelerated by the Tat protein.

[0342] The processes of entry into target host cells, reverse transcription, integration, and protein maturation can be completed in less than 24 hours, and progeny viral particles have been detected within 12 hours of infection. The first progeny viral particles can be released from infected cells approximately 24 hours after infection. Infected T cells are typically eliminated by the immune system (e.g., by cytotoxic T cells) at a rate of 2–4 days. While HIV-infected T cells are destroyed, T helper cells decline due to limited T cell production. The proteins nef and tat also inhibit the maturation and replacement of helper T cells. Consequently, HIV infection eventually leads to immunodeficiency (GAC, Transfusion Medicine Hemotherapy, 43:203–222, 2016, incorporated herein by reference).

[0343] C. Transmission and Pathology HIV enters the body through intact mucous membranes, broken skin, or parenteral inoculation. It is most commonly transmitted sexually. Once infected, HIV can be detected systemically within approximately 10–14 days, and transmission via blood or transplanted organs is possible approximately 5–6 days after infection. Clinical symptoms typically appear 3–6 weeks after infection and may include fever, enlarged lymph nodes, fatigue, rash, gastrointestinal symptoms, acute neuropathy, muscle pain, and / or malaise. However, some individuals are asymptomatic during this acute phase. These symptoms of acute or primary infection may persist for 2–6 weeks. This initial symptomatic phase is then typically followed by an asymptomatic or occasionally symptomatic period, which may last for several years.

[0344] If left untreated, HIV infection causes a progressive loss of CD4+ T cells, which can lead to a series of immunological abnormalities and an increased risk of infectious and oncological complications. In addition, HIV infection is associated with cardiovascular disease, bone disease, renal and liver dysfunction, and several other common morbidities.

[0345] Antiviral therapy (ART) has been developed to treat HIV infection, but ART can only prevent new cells from becoming infected; that is, ART cannot eliminate infection if the cell already contains viral DNA integrated into its genome. Furthermore, in CD4+ T cells, HIV establishes a latent infection that can persist indefinitely, some of which have the ability to self-renew. Once integrated into a cell's genome, HIV can continue to replicate. (Deeks et al., Nature reviews 1.1 2015, and GAC, Transfusion Medicine Hemotherapy, 43:203-222, 2016, which are incorporated herein by reference.)

[0346] D. Treatment strategy The development of therapeutics 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, found worldwide, and HIV-2, found primarily in West Africa). However, HIV-1 is further subdivided into three subgroups (M, N, O, and P), and M is further subdivided into subtypes A to L. Subtypes can also recombine during co-infection, resulting in other, further recombinant subtypes.

[0347] Another challenging factor is the high mutation rate of HIV in vivo. A recent study quantified the spontaneous mutation rate of the entire HIV-1 genome in DNA sequences from peripheral blood mononuclear cells and found a rate of (4.1 ± 1.7) × 10 per base per cell. -3 revealed an extremely high mutation rate of 100 ng / ml, the highest reported mutation rate 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 epitopes that are conserved across multiple groups and subtypes of the ever-mutating HIV sequence is extremely rare, and the virus has a unique ability to evade the immune system.

[0348] Besides its high mutation rate, HIV poses other challenges to the immune system that make treatment uniquely difficult. Although therapeutic targets on HIV include the HIV envelope protein (HIV Env), HIV Env is highly glycosylated, and therefore Env sites are shielded from therapeutic agents by existing glycans. In addition, Env glycans are derived from the host and can be highly heterogeneous.

[0349] Recent therapeutic strategies involve the use of broadly neutralizing antibodies (bNAbs), antibodies capable of neutralizing diverse international HIV isolates. Such antibodies have been identified in HIV-infected individuals considered "elite neutralizers," who comprise less than 10% of HIV patients (Burton and Hangartner, Ann. Rev. Immunol. 2016, incorporated herein by reference). Such antibodies provide insight into potential target epitopes and the structure of therapeutics. Other advances that have aided in the advancement of therapeutics include the generation of stable HIV Env spike trimers (Sanders and Moore, Immunol. Rev. 2017, incorporated herein by reference) and the high-resolution characterization of their structure (Ward and Wilson, Immunol. Rev. 2017, incorporated herein by reference). Examples of potential Env target sites include the proximal end, the high-mannose patch of the gp120 region, the gp120-gp41 interface region, the gp41 juxtamembrane region (MPER), and the CD4-binding site (see Figure 2, originally published by McCoy and Burton, Immunol Rev. 275.1 11-20 2017, incorporated herein by reference). Each of these sites faces unique challenges as a therapeutic target for bNAbs. For example, bnAbs targeting the gp41-gp120 interface must be able to bind complex and heterogeneous glycans. BnAbs targeting the CD4-binding site of the Env protein have been shown to exhibit high levels of somatic hypermutation.

[0350] Nevertheless, among these sites, the proximal site is of particular interest because it is a conserved region identified in the HIV envelope trimer (Env).

[0351] As discussed herein, the HIV-1 envelope (Env) glycoprotein trimer, a trimer of gp120-gp41 heterodimers, is the only viral protein on the surface of the HIV-1 virion and is therefore an important target for bnAbs. The proximal end of the Env trimer contains the variable 1 and 2 (V1V2) loops and the variable 3 (V3) loop. The V1V2 loop contains four β-strands. In addition, the V1V2 epitope is particularly interesting as a target because it is a region of gp120 that undergoes a large conformational change upon CD4 binding. For example, the interaction of CD4 with gp120 stabilizes an open Env conformation capable of interacting with chemokine receptors, triggering further conformational changes that ultimately result in the insertion of the gp41 fusion protein into the target cell membrane. In the closed state, the V1V2 regions from the three gp120 fragments shield V3 and the coreceptor-binding site. For this reason, certain tip-directed bNAbs (e.g., those targeting V1V2 epitopes) are characterized by their ability to prevent the Env trimer from opening and exposing the V3 loop and coreceptor binding site, thereby blocking the conformational change that leads to fusion of the viral membrane with the host cell membrane (Wang, Haoqing, et al., "Asymmetric recognition of HIV-1 Envelope trimer by V1V2 loop-targeting antibodies." Elife 6(2017):e27389, which is incorporated herein by reference).

[0352] The present disclosure provides, among other things, polyribonucleotides that encode antibody agents, e.g., bNABs, that target a broader set of HIV variants and therefore can treat a greater number of HIV patients. Additionally, the present disclosure provides compositions for the delivery of antibody agents, e.g., bNABs, that target various HIV sequences.

[0353] 1. Antiviral treatment for HIV HIV infection is currently primarily treated with antiretroviral therapy (ART). ART is a class of medications that can reduce HIV replication, increase CD4 cell counts, and reduce the risk of transmission in infected individuals. The World Health Organization (WHO) recommends initiating ART in all HIV-infected adults, 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, incorporated herein by reference). However, ART is not a curative therapy, and viremia (e.g., viral load) will soon recur if an infected individual stops taking ART. The high mutation rate of HIV also constrains patients to strictly adhere to treatment to avoid the emergence of escape mutants and treatment failure. Thus, ART is intended to be taken daily throughout the infected subject's life.

[0354] There are several classes of FDA-approved ARTs for treating HIV that act through different mechanisms. Effective management of HIV infection often requires a combination of at least three ARTs to address the complex pathogenesis of the disease. The most effective combination of ARTs often varies among infected individuals (see, e.g., Bhatti et al., Cureus 2016, which is incorporated herein by reference). Cihlar et al., Current opinion in virology, 2016, which is incorporated herein by reference in its entirety, provides an overview of classes of ART drugs for the treatment of HIV. [Table 1]

[0355] 2. HIV antibody drugs In addition to ART, anti-HIV antibodies have been developed. To be used to treat HIV, the antibodies generally must possess certain properties, including safety, a favorable pharmacokinetic profile, highly potent neutralizing activity, and broad neutralizing activity to effectively target the diversity present in the HIV virion. As with other HIV treatments (including, for example, ART), viral escape from anti-HIV antibodies presents a significant challenge.

[0356] For example, see Barouch et al., "Infected rhesus macaques with SHIV-SF162P3" (Barouch, et al., Nature 503:7475 224-228, 2013), which is incorporated herein by reference in its entirety. Rhesus macaques were 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 10 mg / kg cocktail on days 0 and 7, as a 10 mg / kg cocktail on day 0 only, or as a combination of 10 mg / kg PGT121 and 3BNC117 alone. Transient viral suppression was observed until bNAb levels fell below 10 μg / mL. The mAb was also administered alone to monkeys, and PGT121 alone resulted in rapid virologic suppression, which rebounded in most animals after 6–8 weeks. Monkeys receiving the PGT121 / 3BNC117 combination were given a second dose on day 105, after viral levels had rebounded. Virologic suppression was observed again, but suppression was less durable than with the previous dose.

[0357] Shingai, et al. AD8EO(Shingai, et al., Nature 503:7475 277-280, 2013, incorporated herein by reference in its entirety). Here, rhesus macaques were treated with 10-1074 and 3BNC117 mAbs, either alone or in combination. When administered alone at 10 mg / kg 12 weeks postinoculation, both antibodies induced rapid viral suppression, but viral levels quickly rebounded. Administration of both antibodies in combination to chronically infected animals resulted in a longer period of suppression and improved CD4+ T cell levels, but viral levels subsequently rebounded. In other studies, either antibody was pretreated alone in monkeys and found to prevent viral disease. Single-genome analysis of rebound virus in 10-1074-treated monkeys revealed a mutation that eliminated the N332 glycan of gp120, conferring resistance to the mAb. On the other hand, SGA analysis of rebound virus in monkeys treated with both 10-074 and 3BNC117 revealed that not all monkeys contained virus with changes that conferred mAb resistance.

[0358] Caskey et al. described a first-in-human, dose-escalation, Phase 1 clinical trial of 3BNC117 (a CD4 binding site antibody) (Caskey et al., Nature 522.7557:487-491, 2015, which is incorporated herein by reference in its entirety). The trial enrolled uninfected and HIV-1-infected individuals. 3BNC117 at doses of 1, 3, 10, or 30 mg / kg was generally safe and well-tolerated, with no grade 3, 4, or serious adverse events. HIV-1-infected individuals exhibited a faster rate of antibody clearance than uninfected control subjects. The effect of treatment on viral load was dose-dependent, with doses of 10 and 30 mg / kg resulting in a maximum 2.5-log reduction in viral load. Viral resistance was observed in some individuals, but not others, regardless of mAb dose. Viruses were cloned and sequenced, and G459D was the common mutation in the 10 mg / kg group, while the other groups showed a longer V5 loop (other mutations noted). Both mutations may alter sensitivity to anti-CD4b.

[0359] Caskey et al. also evaluated 10-074, a highly potent mAb targeting 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 in 14 uninfected and 19 HIV-1-infected individuals. A single intravenous infusion was administered at 3, 10, or 30 mg / kg. The mAb was generally found to be safe and well-tolerated, with no grade 3, 4, or serious adverse events. HIV-1-infected individuals exhibited a faster rate of antibody clearance than uninfected controls. Treatment suppressed viral load in individuals with 10-074-susceptible strains, followed by rebound. Single genome sequencing (SGS) of the rebound virus revealed that all patients who responded to treatment had PNGS at position N332 and no change. 324 G(D / N)IR 327 Four weeks after injection, 91% of the envelope sequences contained amino acid mutations, 97% of which eliminated PNGS at position 332 by mutating either N332 or S334. 3% of the mutated sequences 324 G(D / N)IR 327 The mutations at D / N325 in the motif were observed. The majority of mutations at the nucleic acid level were rearrangements, consistent with reverse transcriptase errors. Neutralization assays demonstrated that HIV-1 mutants resistant to 10-074 were not resistant to 3BNC117, VRC01, or PGDM1400 (mAbs targeting other regions of HIV-1). SGS performed 1 week after injection revealed that resistance variants were already present or rapidly generated.

[0360] Bar et al. conducted two open-laboratory studies to evaluate the safety, side effect profile, pharmacokinetic properties, and antiviral activity of VRC01 (a bNAb targeting the CD4 binding site of HIV) in patients undergoing antiretroviral therapy (ART) interruption (Bar et al., New England Journal of Medicine 375.21:2037-2050, 2016, incorporated herein by reference in its entirety). One study administered three 40 mg / kg infusions over a 6-week period, while the other administered eight 40 mg / kg infusions over a 6-month period. Treatment was well tolerated, with no grade 3 or higher adverse events. Neither study achieved sustained suppression of plasma viremia, yet slightly prolonged time to rebound compared with historical controls. Regardless of time to rebound, resistance to VRC01 developed in nearly all participants in one study. Viral isolates were more resistant to VRC01 neutralization in pre-treatment versus post-treatment samples. Treatment with VRC01 did not affect susceptibility to neutralization by other bNAbs.

[0361] Mendoza et al. conducted a phase 1b clinical trial evaluating the combination of 3BNC117 and 10-1074, infused at 30 mg / kg doses at weeks 0, 3, and 6 (Mendoza et al., Nature 561.7724:479-484, 2018, 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 reported serious adverse events. The median time to rebound was significantly prolonged with combination bNAb treatment. The two individuals with the fastest rebound were found to have pre-existing strains resistant to one or the other bNAb. Rebound virus was concentrated within a low-diversity lineage, consistent with the proliferation (escape) of one or two relapse viruses. Most of the rebound viruses were found to contain the 10-1074 mutation as opposed to the 3BNC117 mutation. Nevertheless, combination bNAb therapy proved more effective than single bNAb treatment in abrogating viral escape.

[0362] Gautam, et al. AD8EORhesus macaques infected with 3BNC117-LS and treated with 3BNC117-LS and 10-074-LS mAbs were evaluated (Gautam, Rajeev, et al., Nature Medicine 24.5:610-616, 2018, incorporated herein by reference in its entirety). M428L and N343S (collectively referred to as LS) are mutations in the fragment domain of the mAb that extend its half-life. The LS mutations had no effect on virus neutralization in in vitro assays. The LS mAb was administered alone at 20 mg / kg and was well tolerated in all monkeys. Compared with 3BNC117-LS recipients, 10-1074-LS recipients showed enhanced protection against viral challenge, although the LS mutations were more effective than the wild-type in both antibodies. 10-1074-LS decayed in serum at a slower rate than 3BNC117-LS. The concentration / neutralizing activity of mAbs was determined to be predictive of the probability of infection. Only experiments in which antibody pretreatment was followed by virus challenge were performed.

[0363] Schommers et al. characterized an anti-HIV antibody called "1-18" in in vitro assays and in HIV-1-infected humanized mice. It was reported that 1-18 binds to the CD4 binding site of HIV and has strong potency and broad spectrum 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 broad spectrum: (1) 1-18 contains an aromatic residue that mimics CD4 residue Phe43 to target the "Phe43 gp120 pocket," a feature previously reported for anti-HIV antibody N6; (2) 1-18 contacts the adjacent gp120 promoter with an increased buried surface area (due to the insertion of six residues in CDRH1), as previously observed for anti-HIV antibody 3BNC117; and (3) it has a larger buried surface area on gp120 compared to other anti-HIV antibodies. In addition, 1-18 was reported to contact conserved residues on HIV gp120 that other anti-HIV antibodies did not. Schommers hypothesized that these contacts could make 1-18 less reliant on contacts with the classical CD4 binding site, thereby making viral escape more difficult. Nevertheless, Schommers acknowledged that a small number of HIV strains are resistant to 1-18.

[0364] Sok et al. reported that the PGDM1400 antibody showed exceptional potency (median IC50 of 0.003 μg / ml) against a panel of 106 viruses compared to other bNAbs (e.g., PGT121, PGT128, and PGT151). In addition, the PGDM1400 antibody has been shown to have a high neutralization spectrum (83% neutralization rate) (see Sok, Devin, et al., "Recombinant HIV envelope trimer selects for quaternary-dependent antibodies targeting the trimer apex," Proceedings of the National Academy of Sciences 111.49 (2014): 17624-17629; van der Velden, Yme U., et al. "Diverse HIV-1 escape pathways from broadly neutralizing antibody PGDM1400 in humanized mice," Mabs. 12.1 (2020) e1845908, which are incorporated herein by reference in their entireties).

[0365] Taken together, the above data suggest that administration of antibodies may be effective in treating or preventing HIV. However, the difficulty of targeting such mutable viruses is highlighted by the above studies, as well as by the fact that (1) the use of a single broadly neutralizing antibody (bNAb) in therapy leads to the development of HIV resistance to therapy within a few 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) that specific antibody combinations resulted in improved viral control by preventing the early development of resistance (Bar-On et al.This is evident from studies showing the 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. The viral rebound observed with some of these antibodies suggests that they may only be effective for a limited period of time, e.g., before HIV escape mutations begin to develop.

[0366] Thus, there remains a need for therapeutic and prophylactic therapies that can avoid viral escape and remain effective in neutralizing HIV. As described herein, the present disclosure provides technologies useful for administering to a subject one or more antibody agents, e.g., polyribonucleotides encoding anti-HIV antibody agents. Using the technologies and procedures described herein, it is possible, for example, to simultaneously produce different antibody agents from polyribonucleotides. The antibody agent formats are designed to minimize or eliminate the risk of immunoglobulin chain mispairing. The ability to combine multiple antibody agent formulations described herein (e.g., those containing the PGDM1400 antibody agent) allows for the development of compositions (e.g., pharmaceutical compositions) in which multiple antibody agents are delivered together so that they can bind to different epitopes on the HIV virus, thereby minimizing mutation-induced viral escape and improving overall efficacy.

[0367] II. Polyribonucleotides for delivery of antibody agents The present disclosure utilizes, among other things, RNA technology as a modality for direct expression in a subject of antibody agents, a novel class of antibody-based therapeutics. In some embodiments, the polyribonucleotides described herein encode immunoglobulin chains of the antibody agents.

[0368] In some embodiments, the antibody agent targets HIV. In some embodiments, the antibody agent that targets HIV specifically binds to a particular epitope of an HIV polypeptide. For example, in some embodiments, the antibody agent specifically binds to an epitope encompassing the Env trimer head or a portion thereof. See Figure 2, originally from McCoy and Burton, Immunol Rev. 275.1 11-20, 2017, incorporated herein by reference.

[0369] In some embodiments, the antibody agent has a binding affinity (e.g., as measured by a dissociation constant) for an HIV epitope (e.g., an epitope at the Env trimer tip) of 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 -9M or less. In some embodiments, the HIV antibody agent selectively binds to a target epitope of HIV such that the binding between the HIV antibody agent and the target epitope is more than 2-fold, more than 5-fold, more than 10-fold, or more than 100-fold greater than the binding of the HIV antibody agent to a non-target epitope. In some embodiments, the HIV antibody agent may have binding affinity for an HIV epitope and also for variants of the HIV epitope. Those skilled in the art will appreciate that, in some cases, binding affinity (e.g., as measured by a dissociation constant) can be affected by non-covalent intermolecular interactions between two molecules, such as hydrogen bonding, electrostatic interactions, hydrophobic forces, and van der Waals forces. Alternatively, or in addition, the binding affinity between a ligand and its target molecule can be affected by the presence of other molecules. Those of skill in the art will be familiar with a variety of techniques for measuring binding affinity and / or dissociation constants in accordance with the present disclosure, including, but not limited to, ELISA, gel shift assays, pull-down assays, equilibrium dialysis, analytical ultracentrifugation, surface plasmon resonance (SPR), biolayer interferometry, grating-binding interferometry, and spectroscopic assays.

[0370] In some embodiments, antibody agents targeting HIV may comprise or be derived from broadly neutralizing antibodies (bNAbs). In some embodiments, the antibody agent targeting HIV is selected from the group consisting of Sok, et al., PNAS 111.49:17624-17629, 2014; van der Velden, Yme U., et al., Mabs. 12.1:e1845908, 2020; Barouch, et al., Nature 503:7475 224-228, 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 by reference in their entirety for the purposes described herein.

[0371] In some embodiments, an antibody agent targeting HIV can be, for example, 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, PGDM1400, fragments thereof, or combinations 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, fragments thereof, or combinations thereof. For example, in some embodiments, a polyribonucleotide described herein can comprise one or more heavy chain complementarity 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, a polyribonucleotide 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 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. For example, in some embodiments, polyribonucleotides described herein can comprise one or more light chain complementarity 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, a polyribonucleotide 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 described herein may comprise a light chain variable domain from 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S, or PGDM1400.

[0372] In some embodiments, two or more antibody agents (e.g., 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S or PGDM1400, or fragments or variants thereof) can be delivered (e.g., by administration to a subject) using multiple polyribonucleotides, each encoding an immunoglobulin chain of an antibody agent. In some embodiments, three or more antibody agents (e.g., 1-18, PGT121, 3BNC117, b12, 10-1074, 10E8, 10E8v4, VRC01, VRC07-523-L / S or PGDM1400, or fragments or variants thereof) can be delivered (e.g., by administration to a subject) using multiple polyribonucleotides, each encoding an immunoglobulin chain of an antibody agent. In some embodiments, multiple polyribonucleotides, each encoding an immunoglobulin chain of an antibody agent, can be used to deliver (e.g., by administration 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 fragments or variants thereof). In some embodiments, multiple polyribonucleotides, each encoding an immunoglobulin chain of an antibody agent, can be used to deliver (e.g., by administration 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 fragments or variants thereof). In some embodiments, the antibody agents encoded by one or more polyribonucleotides described herein comprise all or a portion of a PGDM1400 antibody.

[0373] An antibody agent described herein (e.g., a PGDM1400 antibody agent) may be characterized by the amino acid sequence of one or more domains in its antibody structure. For example, an antibody agent may comprise at least one heavy (H) chain and at least one light (L) chain interconnected, for example, by a disulfide bond. Each H chain comprises a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. Each light chain comprises a light chain variable region (abbreviated herein as VL) and a light chain constant region. The variable regions of each light / heavy chain (VL / VH) pair form an antigen-binding domain.

[0374] Within each light or heavy chain variable domain are three short segments called complementarity-determining regions ("CDRs"). The six CDRs in an antibody variable domain (three in the light chain variable domain and three in the heavy chain variable domain) fold together in three-dimensional space to form the actual antibody binding site. The terms "LCDR1," "LCDR2," and "LCDR3" provided herein refer to complementarity-determining regions (CDRs) 1, 2, and 3 of the variable light (L) chain of an antibody agent. In some embodiments, a light chain variable domain provided herein comprises, from N-terminus to C-terminus, LCDR1, LCDR2, and LCDR3. Similarly, the terms "HCDR1," "HCDR2," and "HCDR3" provided herein refer to complementarity-determining regions (CDRs) 1, 2, and 3 of the variable heavy (H) chain of an antibody agent. In certain embodiments, a heavy chain variable domain provided herein comprises, from N-terminus to C-terminus, HCDR1, HCDR2, and HCDR3.

[0375] Also present within the variable region are regions called framework regions ("FR"), although they are not contained within the CDRs. Thus, the complementarity-determining regions (CDRs) are distributed between the framework regions. Each VH and VL therefore contains three CDRs and four FRs, which are arranged from the amino terminus to the carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. In general, framework regions are more conserved than variable regions across naturally occurring antibodies.

[0376] PGDM1400 antibodies, including their nucleotide and amino acid sequences, are disclosed in WO2021 / 087015 and U.S. Pat. No. 10,093,720, which are incorporated by reference in their entireties.

[0377] In some embodiments, a PGDM1400 antibody agent comprises HCDR1, HCDR2, and HCDR3 of a heavy chain variable domain having the amino acid sequence of SEQ ID NO: 36. In some embodiments, a PGDM1400 antibody agent comprises LCDR1, LCDR2, and LCDR3 of a light chain variable domain having the amino acid sequence of SEQ ID NO: 43.

[0378] The locations of the CDRs and framework regions within the VH and VL domains of an antibody agent described herein (e.g., a PGDM1400 antibody agent) can be determined using various numbering systems known in the art, such as Kabat, Chothia, AbM, and IMGT (e.g., Johnson et al., Nucleic Acids Res., 29:205-206 (2001); Chothia and Lesk, J. Mol. Biol., 196:901-917 (1987); Chothia et al., Nature, 342:877-883 (1989); Chothia et al., J. Mol. Biol., 227:799-817 (1992); Al-Lazikani et al., Nature, 342:877-883 (1989); Chothia et al., J. Mol. Biol., 227:799-817 (1992); Al-Lazikani et al., Nature, 342:877-883 (1989); al., J. Mol. Biol., 273:927-748 (1997) ImMunoGenTics (IMGT) numbering; Lefranc, M.-P., The Immunologist, 7, 132-136 (1999); Lefranc, MP et al., Dev. Comp. Immunol., 27, 55-77 (2003), each of which is incorporated herein by reference.) Thus, CDRs within a PGDM1400 antibody agent within the same VH or VL domain may be determined by different numbering systems.

[0379] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of a heavy chain variable domain represented by the amino acid sequence of SEQ ID NO: 36, according to the Kabat numbering system. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of a heavy chain variable domain represented by the amino acid sequence of SEQ ID NO: 36, according to the Chothia numbering system. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of a heavy chain variable domain represented by the amino acid sequence of SEQ ID NO: 36, according to the IMGT numbering system. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences of a heavy chain variable domain represented by the amino acid sequence of SEQ ID NO: 36, according to the AbM numbering system.

[0380] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of a light chain variable domain represented by the amino acid sequence of SEQ ID NO: 43, according to the Kabat numbering. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of a light chain variable domain represented by the amino acid sequence of SEQ ID NO: 43, according to the Chothia numbering. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of a light chain variable domain represented by the amino acid sequence of SEQ ID NO: 43, according to the IMGT numbering. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences of a light chain variable domain represented by the amino acid sequence of SEQ ID NO: 43, according to the AbM numbering.

[0381] In some embodiments, HCDR1 of an antibody agent (eg, a PGDM1400 antibody agent) comprises the amino acid sequence set forth in SEQ ID NO: 18, 1484, or 1490.

[0382] In some embodiments, HCDR2 of an antibody agent (eg, a PGDM1400 antibody agent) comprises the amino acid sequence set forth in SEQ ID NO: 21, 1485, 1491, or 1492.

[0383] In some embodiments, the HCDR3 of an antibody agent (eg, a PGDM1400 antibody agent) comprises the amino acid sequence set forth in SEQ ID NO: 24, 1486, or 1493.

[0384] In some embodiments, LCDR1 of an antibody agent (eg, a PGDM1400 antibody agent) comprises the amino acid sequence set forth in SEQ ID NO:27 or 1487.

[0385] In some embodiments, LCDR2 of an antibody agent (eg, a PGDM1400 antibody agent) comprises the amino acid sequence set forth in SEQ ID NO:30 or 1488.

[0386] In some embodiments, the LCDR3 of an antibody agent (eg, a PGDM1400 antibody agent) comprises the amino acid sequence set forth in SEQ ID NO: 33 or 1489.

[0387] In some embodiments, an antibody agent (eg, a PGDM1400 antibody agent) comprises one or more CDRs, or any set of CDRs, shown in Table 2 below. [Table 2-1] [Table 2-2]

[0388] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 1484, 1492, and 1486, respectively. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 1490, 1491, and 1486, respectively. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 18, 21, and 1493, respectively.

[0389] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth, respectively, in SEQ ID NOs: 18, 21, and 24. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth, respectively, in SEQ ID NOs: 1484, 1485, and 1486.

[0390] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 1487, 1488, and 1489, respectively. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 27, 30, and 1489, respectively.

[0391] In some embodiments, an antibody agent (eg, a PGDM1400 antibody agent) comprises the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 27, 30, and 33, respectively.

[0392] In some embodiments, the antibody agent (e.g., a PGDM1400 antibody agent) comprises (i) the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 18, 21, and 24, respectively; and (ii) the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 27, 30, and 33, respectively.

[0393] In some embodiments, the antibody agent (e.g., a PGDM1400 antibody agent) comprises (i) the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 1484, 1485, and 1486, respectively; and (ii) the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 1487, 1488, and 1489, respectively.

[0394] In some embodiments, the antibody agent (e.g., a PGDM1400 antibody agent) comprises (i) the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 1490, 1491, and 1486, respectively; and (ii) the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 1487, 1488, and 1489, respectively.

[0395] In some embodiments, the antibody agent (e.g., a PGDM1400 antibody agent) comprises (i) the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 1484, 1492, and 1486, respectively; and (ii) the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 1487, 1488, and 1489, respectively.

[0396] In some embodiments, the antibody agent (e.g., a PGDM1400 antibody agent) comprises (i) the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 1490, 1491, and 1486, respectively; and (ii) the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 1487, 1488, and 1489, respectively.

[0397] In some embodiments, the antibody agent (e.g., a PGDM1400 antibody agent) comprises (i) the HCDR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 18, 21, and 1493, respectively; and (ii) the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 27, 30, and 1489, respectively.

[0398] In some embodiments, a PGDM1400 antibody agent comprises the FR1, FR2, FR3, and / or FR4 domains of a heavy chain variable domain represented by the amino acid sequence of SEQ ID NO: 36. In some embodiments, a PGDM1400 antibody agent comprises the FR1, FR2, FR3, and / or FR4 domains of a light chain variable domain represented by the amino acid sequence of SEQ ID NO: 43.

[0399] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a heavy chain FR1 comprising the amino acid sequence set forth in SEQ ID NO: 1496. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a heavy chain FR2 comprising the amino acid sequence set forth in SEQ ID NO: 1497. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a heavy chain FR3 comprising the amino acid sequence set forth in SEQ ID NO: 1498. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a heavy chain FR4 comprising the amino acid sequence set forth in SEQ ID NO: 1499.

[0400] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a light chain FR1 comprising the amino acid sequence set forth in SEQ ID NO: 1500. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a light chain FR2 comprising the amino acid sequence set forth in SEQ ID NO: 1501. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a light chain FR3 comprising the amino acid sequence set forth in SEQ ID NO: 1502. In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) comprises a light chain FR4 comprising the amino acid sequence set forth in SEQ ID NO: 1503.

[0401] In some embodiments, the FR1, FR2, FR3 and / or FR4 domains of the heavy chain of an antibody agent (e.g., a PGDM1400 antibody agent) comprise the amino acid sequences set forth in SEQ ID NOs: 1496, 1497, 1498 and 1499, respectively. In some embodiments, the FR1, FR2, FR3 and / or FR4 domains of the light chain of an antibody agent (e.g., a PGDM1400 antibody agent) comprise the amino acid sequences set forth in SEQ ID NOs: 1500, 1501, 1502 and 1503, respectively.

[0402] In some embodiments, an antibody agent (eg, a PGDM1400 antibody agent) comprises the HCDR1, FR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 18, 1496, 21, and 24, respectively.

[0403] In some embodiments, the antibody agent (e.g., a PGDM1400 antibody agent) comprises: (i) a VH domain comprising the HCDR1, FR1, HCDR2, and HCDR3 amino acid sequences set forth in SEQ ID NOs: 18, 1496, 21, and 24, respectively; and (ii) a VL domain comprising the LCDR1, LCDR2, and LCDR3 amino acid sequences set forth in SEQ ID NOs: 27, 30, and 33, respectively.

[0404] In some embodiments, an antibody agent encoded by one or more polyribonucleotides provided herein comprises all or a portion of a PGDM1400 antibody. In some embodiments, the antibody agent comprises a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof. In some embodiments, the antibody agent comprises a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24). In some embodiments, the antibody agent comprises a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. In some embodiments, the antibody agent comprises a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33). In some embodiments, the antibody agent comprises: (a) a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof; and (b) a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. In some embodiments, the antibody agent comprises: (a) a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24); and (b) a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33).

[0405] In some embodiments, the polyribonucleotides described herein encode all or a portion of a PGDM1400 antibody. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising a heavy chain variable domain, wherein the heavy chain variable domain comprises (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (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 comprises (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), 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 (SEQ ID NO:27), (ii) LCDR2 (LAS, SEQ ID NO:30), and (iii) LCDR3 (SEQ ID NO:33). 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 (SEQ ID NO:18), (ii) HCDR2 (SEQ ID NO:21), (iii) HCDR3 (SEQ ID NO:24), or (iv) a combination thereof; and the light chain variable domain comprises (i) LCDR1 (SEQ ID NO:27), (ii) LCDR2 (LAS, SEQ ID NO:30), (iii) LCDR3 (SEQ ID NO:33), or (iv) a combination thereof.In some embodiments, the polyribonucleotides described herein encode immunoglobulin chains comprising a heavy chain variable domain and a light chain variable domain, wherein the heavy chain variable domain comprises (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24); and the light chain variable domain comprises (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33). In some embodiments, the polyribonucleotides described herein encode two immunoglobulin chains: a first immunoglobulin chain comprising a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof; and a second immunoglobulin chain comprising a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), 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 comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24); and a second immunoglobulin chain comprising a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33).

[0406] In some embodiments, an 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 set forth in SEQ ID NO: 36. In some embodiments, an 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% identity to the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, an antibody agent comprises a heavy chain variable domain set forth in SEQ ID NO: 36. In some embodiments, an antibody agent comprises a light chain variable domain set forth in SEQ ID NO: 43.

[0407] In some embodiments, an antibody agent (e.g., a PGDM1400 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 set forth in SEQ ID NO: 1494. In some embodiments, an antibody agent (e.g., a PGDM1400 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% identity to the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, an antibody agent comprises a heavy chain variable domain set forth in SEQ ID NO: 1494. In some embodiments, an antibody agent comprises a light chain variable domain set forth in SEQ ID NO: 43.

[0408] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain comprising one or more mutations compared to the amino acid sequence set forth in SEQ ID NO: 1494. In some embodiments, there is a substitution mutation at residues 3 and / or 5 compared to SEQ ID NO: 1494. In some embodiments, the substitution mutation at residue 3 compared to SEQ ID NO: 1494 results in the presence of a positively charged amino acid at residue 3. Positively charged amino acids include Lys (K), Arg (R), and His (H). In some embodiments, the substitution mutation at residue 3 compared to SEQ ID NO: 1494 includes Q3H. In some embodiments, the substitution mutation at residue 5 compared to SEQ ID NO: 1494 results in the presence of a polar amino acid at residue 5. Polar amino acids include Ser (S), Thr (T), Tyr (Y), Asn (N), and Gln (Q). In some embodiments, the substitution mutation at residue 5 compared to SEQ ID NO: 1494 includes V5T.

[0409] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain comprising SEQ ID NO: 1494 with substitution mutations Q3H and V5T.

[0410] In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising 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 set forth in SEQ ID NO: 36. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain comprising 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 set forth in SEQ ID NO: 43. In some embodiments, the polyribonucleotides described herein encode immunoglobulin chains 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 set forth in SEQ ID NO: 36, and 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 set forth in SEQ ID NO: 43.

[0411] In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a heavy chain variable (VH) domain. In some embodiments, the VH domain comprises the VH domain of the PGDM1400 antibody. In some embodiments, the polyribonucleotides encode the VH domain of an antibody (e.g., as described herein) selected from PGT121, 3BNC117, b12, 10-1074, 10-1074-LS, 10E8, VRC01, VRC07-523, or 1-18.

[0412] In some embodiments, the polyribonucleotide comprises a VH domain coding sequence comprising: (a) an HCDR1 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 19, (b) an HCDR2 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 22, (c) an HCDR3 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 25, or (d) a combination thereof. In some embodiments, the polyribonucleotide comprises a VH domain coding sequence comprising: (a) an HCDR1 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 19, (b) an HCDR2 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 22, and (c) an HCDR3 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO: 25. 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: 37, 39 or 41. In some embodiments, the polyribonucleotide encodes a VH domain and comprises a VH coding sequence according to SEQ ID NO: 37, 39 or 41.

[0413] In some embodiments, the polyribonucleotide described herein comprises an immunoglobulin chain of an antibody agent, wherein the immunoglobulin chain comprises a variable light (VL) domain. In some embodiments, the VL domain comprises the VL domain of the PGDM1400 antibody. In some embodiments, the polyribonucleotide encodes the VL domain of an antibody (e.g., one described herein) selected from PGT121, 3BNC117, b12, 10-1074, 10-1074-LS, 10E8, VRC01, VRC07-523, or 1-18.

[0414] In some embodiments, the polyribonucleotide comprises one or more coding regions encoding an immunoglobulin chain of an antibody agent, the immunoglobulin chain comprising 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:28, (b) a LCDR2 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO:31, (c) a LCDR3 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO:34, or (d) a combination thereof. 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:28, (b) a LCDR2 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO:31, and (c) a LCDR3 coding sequence comprising a ribonucleic acid sequence according to SEQ ID NO:34. In some embodiments, the polyribonucleotide encodes a VL domain and comprises a VL coding sequence that is at least 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to SEQ ID NO: 44 or 46. In some embodiments, the polyribonucleotide encodes a VL domain and comprises a VL coding sequence according to SEQ ID NO: 44 or 46.

[0415] In some embodiments, the antibody agent is formed by one, two, three, or four immunoglobulin chains.

[0416] In some embodiments, the polyribonucleotides described herein encode a single immunoglobulin chain. In some embodiments, a first polyribonucleotide encodes a first immunoglobulin chain of an antibody agent. In some embodiments, a first polyribonucleotide encodes a first immunoglobulin chain of an antibody agent, and a second polyribonucleotide encodes a second immunoglobulin chain of an antibody agent. In some embodiments, a first polyribonucleotide encodes a first immunoglobulin chain of an antibody agent, a second polyribonucleotide encodes a second immunoglobulin chain of an antibody agent, and a third polyribonucleotide encodes a third immunoglobulin chain of an antibody agent. In some embodiments, a first polyribonucleotide encodes a first immunoglobulin chain of an antibody agent, a second polyribonucleotide encodes a second immunoglobulin chain of an antibody agent, a third polyribonucleotide encodes a third immunoglobulin chain of an antibody agent, and a fourth polyribonucleotide encodes a fourth immunoglobulin chain of an antibody agent.

[0417] In some embodiments, the polyribonucleotides described herein encode two immunoglobulin chains. In some embodiments, a single polyribonucleotide can include a first coding region encoding a first immunoglobulin chain of an antibody and a second coding region encoding a second immunoglobulin chain of the antibody. In some embodiments, the first and second coding regions are separated by an internal ribosome entry site (IRES), an internal promoter, or a peptide sequence, such as a "self-cleaving" 2A or 2A-like sequence (see, e.g., Szymczak et al. Nat Biotechnol 22:589, May 2004; ePub April 4 2004, which is incorporated herein by reference), to generate the first and second immunoglobulin chains from a single polyribonucleotide.

[0418] Antibody agents encoded by one or more polyribonucleotides described herein can be in a variety of formats as described herein. Exemplary types of antibody agents include, but are not limited to, monoclonal or polyclonal antibodies. In some embodiments, antibody agents can include one or more sequence elements that are humanized, chimerized, etc., as known in the art. In some embodiments, antibody agents utilized in accordance with the present disclosure include, but are not limited to, unmodified IgG, IgA, IgG, IgE, or IgM antibodies; bispecific or multispecific antibodies (e.g., Zybodies®); CrossMabs (e.g., CrossMabs®); CH1-CLx , CrossMab CH1-CLcv Bispecific CrossMAb with Knobs-in-Hole CH1-CLx antibody fragments, e.g., Fab fragments, Fab' fragments, F(ab')2 fragments, Fd' fragments, Fd fragments, and isolated complementarity determining regions (CDRs) or sets thereof; single chain Fv (scFv); scFv-Fc fusions; polypeptide-Fc fusions; single domain antibodies (e.g., shark single domain antibodies, e.g., IgNAR or fragments thereof); camelid antibodies; masked antibodies (e.g., Probody®), small modular immunopharmaceuticals ("SMIP™"); single chain or tandem diabodies (TandAb®) ); VHH; Anticalins®; Nanobodies® minibodies; BiTEs®; Ankyrin repeat proteins or DARPINs®; Avimers®; DARTs; TCR-like antibodies; Adnectins®; Affilins®; Transbodies®; Affibodies®; TrimerX®; Microproteins; Fynomer®, Sentilin®; and KALBITOR®. In some embodiments, immunoglobulin chains and / or fragments of such antibodies may be used in combination, for example, combining scFv-Fc arms with conventional antibody arms.

[0419] Exemplary formats that may be used in accordance with the present disclosure are further described below.

[0420] A. Conventional Antibodies In some embodiments, the polyribonucleotides described herein can be used to express conventional antibodies. As used herein, "conventional antibody" refers to an antibody agent comprising two heavy chains and two light chains (see, e.g., Figure 5, panel A). Each heavy chain comprises 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 comprise 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 comprise a CH1 domain, a hinge domain, a CH2 domain, a CH3 domain, a CH4 domain, or a combination thereof. Each light chain comprises a light chain variable domain operably linked to a light chain constant domain.

[0421] Typically, heavy and light chain variable domains can be further subdivided into regions of variability called complementarity-determining regions (CDRs), separated by more conserved regions called framework regions (FRs). Such heavy and light chain variable domains can each comprise three CDRs and four framework regions, arranged, for example, from amino to carboxyl terminus in the following order: 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 referred to as "HCDR1," "HCDR2," and "HCDR3," respectively, and the CDRs in the light chain are referred to as "LCDR1," "LCDR2," and "LCDR3," respectively.

[0422] The conventional antibodies described herein can comprise any one of the five major classes of antibodies: IgA, IgD, IgE, IgG, and IgM. In some embodiments, the conventional antibodies comprise an IgG or IgA antibody. In some embodiments, the conventional antibodies described herein comprise a particular isotype selected from the group of IgA and IgG isotypes: IgG1, IgG2, IgG3, IgG4, IgA1, and IgA2. Furthermore, in some embodiments, the conventional antibodies can comprise any particular heavy chain constant domain corresponding to the different classes of immunoglobulins, including α, δ, ε, γ, and μ, respectively. In some embodiments, the conventional antibody is an unmodified IgG1 antibody or other antibody class or isotype 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, there is allelic diversity within IgG subclasses, giving rise to allotypic variants or allotypes. The IgG antibody agents described herein may include specific allotypes, 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).

[0423] The Fc region of a conventional antibody binds to elements of the complement system and also binds to receptors on effector cells, e.g., effector cells that mediate cytotoxicity. As is known in the art, the affinity and / or other binding attributes of the Fc region for an Fc receptor can be modulated by glycosylation or other modifications. In some embodiments, conventional antibodies produced and / or utilized in accordance with the present invention comprise a glycosylated Fc domain, including, for example, an Fc domain having altered or engineered glycosylation. In some embodiments, conventional antibodies are naturally produced (e.g., generated by an organism that responds to an antigen) or produced by recombinant engineering, chemical synthesis, or other artificial systems or methodologies. In some embodiments, conventional antibodies are polyclonal; in some embodiments, conventional antibodies are monoclonal. In some embodiments, conventional antibodies have constant region sequences characteristic of murine, rabbit, primate, or human antibodies. In some embodiments, conventional antibody sequence elements are humanized, chimerized, etc., as is known in the art.

[0424] A conventional antibody as described herein is an antibody having a heavy chain with a structure substantially similar to a naturally occurring antibody structure or containing an Fc region as defined herein.

[0425] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides provided herein comprises all or a portion of a PGDM1400 antibody. In some embodiments, the conventional antibody comprises a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof. In some embodiments, the conventional antibody comprises a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24). In some embodiments, the conventional antibody comprises a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. In some embodiments, the conventional antibody comprises a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33). In some embodiments, the conventional antibody comprises: (a) a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof; and (b) a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. In some embodiments, the conventional antibody comprises: (a) a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24); and (b) a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33).

[0426] In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of a conventional antibody, the immunoglobulin chain comprising a heavy chain variable domain, the heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of a conventional antibody, the immunoglobulin chain comprising a heavy chain variable domain, the heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24). In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of a conventional antibody, the immunoglobulin chain comprising a light chain variable domain, the light chain variable domain comprising (i) LCDR1 (SEQ ID NO:27), (ii) LCDR2 (LAS, SEQ ID NO:30), (iii) LCDR3 (SEQ ID NO:33), or (iv) a combination thereof. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of a conventional antibody, the immunoglobulin chain comprising a light chain variable domain, the light chain variable domain comprising (i) LCDR1 (SEQ ID NO:27), (ii) LCDR2 (LAS, SEQ ID NO:30), and (iii) LCDR3 (SEQ ID NO:33). 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 comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof; and a second immunoglobulin chain comprising a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), 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 comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24); and a second immunoglobulin chain comprising a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33).

[0427] 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 set forth in SEQ ID NO: 36. 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% identity to the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, a conventional antibody comprises a heavy chain variable domain set forth in SEQ ID NO: 36. In some embodiments, a conventional antibody comprises a light chain variable domain set forth in SEQ ID NO: 43.

[0428] 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 set forth in SEQ ID NO: 1494. 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% identity to the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, a conventional antibody comprises a heavy chain variable domain set forth in SEQ ID NO: 1494. In some embodiments, a conventional antibody comprises a light chain variable domain set forth in SEQ ID NO: 43.

[0429] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain comprising one or more mutations compared to the amino acid sequence set forth in SEQ ID NO: 1494. In some embodiments, there is a substitution mutation at residues 3 and / or 5 compared to SEQ ID NO: 1494. In some embodiments, the substitution mutation at residue 3 compared to SEQ ID NO: 1494 results in the presence of a positively charged amino acid at residue 3. Positively charged amino acids include Lys (K), Arg (R), and His (H). In some embodiments, the substitution mutation at residue 3 compared to SEQ ID NO: 1494 includes Q3H. In some embodiments, the substitution mutation at residue 5 compared to SEQ ID NO: 1494 results in the presence of a polar amino acid at residue 5. Polar amino acids include Ser (S), Thr (T), Tyr (Y), Asn (N), and Gln (Q). In some embodiments, the substitution mutation at residue 5 compared to SEQ ID NO: 1494 includes V5T.

[0430] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain comprising SEQ ID NO: 1494 with substitution mutations Q3H and V5T.

[0431] In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of a conventional antibody, 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 set forth in SEQ ID NO: 36. In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of a conventional antibody, wherein the immunoglobulin chain 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 set forth in SEQ ID NO: 43.

[0432] A conventional antibody encoded by one or more polyribonucleotides described herein 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 conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH3 domain comprising a G1m3, G1m17, or G1m17,1 allotype. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein 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 set forth in any one of SEQ ID NOs: 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, or 1495. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH3 domain having an amino acid sequence set forth in any one of SEQ ID NOs: 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146, or 1495. In some embodiments, the CH3 domain comprises the amino acid sequence of SEQ ID NO: 1495 with substitution mutations D16E and L18M.

[0433] A conventional antibody encoded by one or more polyribonucleotides described herein may comprise one or more heavy chain constant domains containing amino acid modifications (e.g., substitutions or deletions) at one or more amino acid positions. For example, a conventional antibody encoded by one or more polyribonucleotides described herein may comprise an L / S mutation in the CH3 region (to enhance FcRn binding) (see Zalevsky J et al. Nat Biotechnol. 2010, which is incorporated herein by reference). Such mutations are designated M428L and N434S (i.e., M88L and N94S in the CH3 domain, e.g., SEQ ID NO: 113, 116, or 1495) based on EU numbering, and are referred to herein as "LS" or "L / S" (see, e.g., panel D of Figure 5). In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises an E294 deletion (for over-sialic acid addition to the Fc) (see Bas M et al. J Immunol 2019, which is incorporated herein by reference).

[0434] The present disclosure also provides techniques that can be used to express antibody agents, such as those shown in Figure 3 or described in Stadler et al. (2016) Oncoimmunology 5(3):e1091555 and / or Stadler et al. (2017) Nature Medicine 23(7):815-817. Producing multiple antibody agents from a single composition (e.g., a composition containing sufficient polyribonucleotides to encode multiple antibody agents) presents challenges, particularly because unregulated pairing of different antibody heavy and light chains can result in undesired antibody species. Due to the presence of mismatched by-products and a significant decrease in production yield, sophisticated purification procedures are required to isolate the desired antibody agents under these circumstances (see, e.g., Morrison, SL, Nature Biotech. 25, 1233-1234, 2007, which is incorporated herein by reference). Generally, even when recombinant expression techniques are used, the same problem of mismatched by-products remains. One approach to solving the problem of mispairing by-products is known as the "knob-into-hole" (KIH) technique, which aims to force pairing of two different antibody heavy chains by introducing mutations into the CH3 domain to alter the contact interface. In one chain, bulky amino acids were replaced with amino acids with short side chains to create a "hole," while in the CH3 domain of the other, amino acids with large side chains were introduced to create a "knob." Coexpression of these two heavy chains with two light chains resulted in the formation of heterodimers in higher yields than homodimers (see Ridgway, JB, et al., Protein Eng. 9, 617-621, 1996, and WO 96 / 027011, which are incorporated herein by reference in their entireties). In some embodiments, the antibody agents described herein utilize KIH technology, such as that described in WO 1998 / 050431, which is incorporated herein by reference in its entirety. As described herein, antibody agents may include specific mutations utilizing KIH technology, including but not limited to modifications of CH3.In some embodiments, the antibody agent comprises a CH3 domain comprising one or more of the following mutations: Y349C, T366S, L368A, and Y407V (based on 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 (based on EU numbering). Such combinations of mutations are referred to herein as "cah." In some embodiments, the antibody agent comprises a CH3 domain comprising one or more mutations selected from S354C and T366W (based on EU numbering). In some embodiments, the antibody agent comprises a CH3 domain comprising each of the following mutations: S354C and T366W (based on EU numbering). Such combinations of CH3 mutations are referred to herein as "cak."

[0435] Thus, in some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH3 domain comprising one or more of the following mutations: Y349C, T366S, L368A, and Y407V (based on EU numbering). In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH3 domain comprising one or more mutations selected from S354C and T366W (based on EU numbering).

[0436] In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one of the following substitution mutations: M428L, 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: 117 and 135. In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one or more of the following substitution mutations: Y349C, T366S, L368A, and Y407V (based on EU numbering). In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NOs: 120 and 138. In some embodiments, the polyribonucleotide encodes a CH3 domain comprising one or both of the following substitution mutations: S354C and T366W (based on EU numbering). In some embodiments, the polyribonucleotide comprises a ribonucleic acid sequence according to SEQ ID NOs: 126 and 144.

[0437] 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: 114. In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence encoding a CH3 domain comprising a G1m3, G1m17, or G1m17,1 allotype. In some embodiments, the polyribonucleotide comprises a CH3 ribonucleic acid sequence comprising any one of SEQ ID NOs: 114 and 132.

[0438] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH1 domain. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH1 domain comprising a G1m3 allotype. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH1 domain comprising a G1m17 allotype. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH1 domain comprising the amino acid sequence set forth in SEQ ID NO: 76 or 81.

[0439] 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: 77 or 79. 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: 77, 79, or 82.

[0440] In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more mutations. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising the addition of one or more serine residues. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising the addition of two additional serine residues (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: 91 or 94. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence represented in SEQ ID NO: 91 or 94. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more charge variant mutations. In some embodiments, the polyribonucleotide encodes a CH1 domain comprising one or more substitution mutations selected from 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: 85 or 88. In some embodiments, the polyribonucleotide encodes a CH1 ribonucleic acid sequence according to SEQ ID NO: 85 or 88.

[0441] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a hinge domain. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a hinge domain (referred to herein as "hinge" in Tables 2 and 4) comprising the amino acid sequence set forth in SEQ ID NO: 161.

[0442] In some embodiments, the polyribonucleotide encodes a hinge domain. In some embodiments, the polyribonucleotide encodes a hinge ribonucleic acid sequence represented by SEQ ID NO: 162. In some embodiments, the polyribonucleotide encodes a hinge domain comprising an amino acid modification comprising a deletion of one or more amino acid residues. In some embodiments, the polyribonucleotide encodes a hinge domain comprising an amino acid modification comprising a deletion of amino acid residues EPKSC in a conventional Ig hinge domain (represented by SEQ ID NO: 161). Such a modification is referred to herein as "hinge_del" or "ΔEPKSC." In some embodiments, the polyribonucleotide encodes a hinge ribonucleic acid sequence represented by SEQ ID NO: 168. In some embodiments, the polyribonucleotide encodes a hinge domain comprising an amino acid modification comprising a C220S mutation (based on EU numbering). Such a mutant hinge domain is referred to herein as "hinge_S" or "C / S." In some embodiments, the polyribonucleotide encodes a hinge ribonucleic acid sequence represented by SEQ ID NO: 165.

[0443] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH2 domain. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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 that set forth in SEQ ID NO: 96. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH2 domain having the amino acid sequence set forth in SEQ ID NO: 96.

[0444] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH2 domain with one or more mutations (e.g., with respect to SEQ ID NO: 96). For example, in some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises one or more of the following mutations: G236A, A330L, and I332E (based on EU numbering). For example, in some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises the following mutations, referred to herein as "GAALIE": G236A, A330L, and I332E (based on EU numbering). Such mutations in the CH2 domain are associated with increased affinity for the Fc receptors FcgRIIA and FcgRIII to enhance antibody effector function.

[0445] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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: 99 or 102. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH2 domain having an amino acid sequence set forth in SEQ ID NO: 99 or 102.

[0446] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises one or more mutations selected from G236A and I332E (based on EU numbering). In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a mutation selected from G236A and I332E (based on EU numbering), referred to herein as "GAIE." In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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: 104. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH2 domain having the amino acid sequence set forth in SEQ ID NO: 104.

[0447] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises the mutation G236A (based on EU numbering), referred to herein as "GA." In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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: 107. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH2 domain having the amino acid sequence set forth in SEQ ID NO: 107.

[0448] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises the mutation I332E (based on EU numbering), referred to herein as "IE." In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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: 110. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a CH2 domain having the amino acid sequence set forth in SEQ ID NO: 110.

[0449] 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: 97. 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, referred to herein as "GAALIE": G236A, A330L, and I332E (based on EU numbering). Such mutations in the CH2 domain are associated with increased affinity for the Fc receptors FcgRIIA and FcgRIII to enhance antibody effector function. In some embodiments, the CH2 ribonucleic acid sequence comprises or consists of a sequence according to SEQ ID NO: 100 or 103. In some embodiments, the CH2 ribonucleic acid sequence encodes one or more of the following mutations: G236A and I332E (based on EU numbering), referred to herein as "GAIE." In some embodiments, the CH2 ribonucleic acid sequence comprises the sequence according to SEQ ID NO: 105. In some embodiments, the CH2 ribonucleic acid sequence encodes the mutation G236A (based on EU numbering), referred to herein as "GA." In some embodiments, the CH2 ribonucleic acid sequence comprises the sequence according to SEQ ID NO: 108. In some embodiments, the CH2 ribonucleic acid sequence encodes the mutation I332E (based on EU numbering), referred to herein as "IE." In some embodiments, the CH2 ribonucleic acid sequence comprises the sequence according to SEQ ID NO: 111. In some embodiments, the CH2 ribonucleic acid sequence encodes a CH2 domain comprising an E294 deletion (based on EU numbering).

[0450] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a signal peptide that includes a human signal peptide. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a signal peptide that includes SEQ ID NO:1.

[0451] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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 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: 149. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a kappa light chain constant domain having the amino acid sequence set forth in SEQ ID NO: 149. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises a lambda chain variable domain.

[0452] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises an immunoglobulin chain (e.g., an immunoglobulin 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 a sequence represented by SEQ ID NOs: 170-265, 1305-1306, 1308-1309, or 1314-1315. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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: 170-265, 1305-1306, 1308-1309, and 1314-1315. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides described herein comprise 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 NOs: 842, 843, and 1311-1323. In some embodiments, conventional antibodies encoded by one or more polyribonucleotides described herein comprise an immunoglobulin chain (e.g., an immunoglobulin light chain) encoded by a nucleic acid sequence represented by SEQ ID NOs: 842-843 and 1311-1312.

[0453] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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 set forth in any one of SEQ ID NOs: 1307, 1310, or 1316. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises an immunoglobulin chain (e.g., an immunoglobulin heavy chain) comprising an amino acid sequence set forth in any one of SEQ ID NOs: 1307, 1310, or 1316.

[0454] In some embodiments, a conventional antibody encoded by one or more polyribonucleotides 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: 1313. In some embodiments, a conventional antibody encoded by one or more polyribonucleotides described herein comprises an immunoglobulin chain (e.g., an immunoglobulin light chain) comprising an amino acid sequence set forth in SEQ ID NO: 1313.

[0455] Exemplary immunoglobulin chain (eg, immunoglobulin heavy or light chain) configurations of the conventional antibodies described herein are shown in Table 3 below. [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4] [Table 3-5]

[0456] B.CrossMab CH1-CLx The present disclosure also provides techniques that can be used to deliver and express the antibody agents described herein in a "CrossMab" format (see, e.g., WO2015 / 101588A1, WO2009 / 080253A1, and Schaefer, W. et al., PNAS, 108, 11187-1191, 2011, which are incorporated herein by reference in their entireties). In some embodiments, antibody agents in a CrossMab format have a CL-CH1 cross-link (referred to herein as "CrossMab") in one or both binding arms. CH1-CLx " or "CH1-CLx"). Such modification reduces by-product formation resulting from mispairing of a light chain of a first antibody that specifically binds a first antigen with a heavy chain of a second antibody that specifically binds a second antigen (compared to approaches that do not involve such domain swapping).

[0457] In some embodiments, the antibody agents encoded by one or more polyribonucleotides provided herein comprise a first immunoglobulin chain and a second immunoglobulin chain. In some embodiments, the polyribonucleotides are selected from the group consisting of CrossMab CH1-CLx The polyribonucleotide encoding the first immunoglobulin chain may encode a first immunoglobulin chain and a 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 4, panel B). In some embodiments, the CrossMab CH1-CLx The polyribonucleotide encoding the antibody agent comprises a ribonucleic acid sequence encoding any one of the immunoglobulin chain constructs in Table 4 corresponding to SEQ ID NOs: 266-361, 1317-1318, 1329-1330, 1332-1333, 1335-1336, and 1338-1339. In some embodiments, the CrossMab CH1-CLxThe polyribonucleotide encoding the drug antibody comprises a ribonucleic acid sequence encoding any one of the immunoglobulin chain structures in Table 4 corresponding to SEQ ID NOs: 844-847 and 1230-1321.

[0458] In some embodiments, CrossMab CH1-CLx The antibody agent may be encoded by two separate polyribonucleotides: a first polyribonucleotide comprising coding regions 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 9, panel A), and a second polyribonucleotide comprising coding regions encoding (in 5' to 3' order) a light chain variable domain (VL) and a CH1 domain (see, e.g., Figure 9, panel B).

[0459] In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The antibody agent comprises all or a portion of a PGDM1400 antibody. CH1-CLx The antibody agent comprises a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof. CH1-CLx The antibody agent comprises a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24). CH1-CLx The antibody agent comprises a light chain variable domain comprising: (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. CH1-CLx The antibody agent comprises a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33). CH1-CLxThe antibody agent comprises: (a) a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof; and (b) a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. In some embodiments, the CrossMab CH1-CLx The antibody agent comprises: (a) a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24); and (b) a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33).

[0460] In some embodiments, the polyribonucleotides described herein are CH1-CLx In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of an antibody agent, the immunoglobulin chain comprising a heavy chain variable domain, the heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof. CH1-CLx In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of an antibody agent, the immunoglobulin chain comprising a heavy chain variable domain, the heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24). CH1-CLx In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of an antibody agent, the immunoglobulin chain comprising a light chain variable domain, the light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. CH1-CLxIn some embodiments, the polyribonucleotides described herein encode immunoglobulin chains of an antibody agent, the immunoglobulin chains comprising a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33). In some embodiments, the polyribonucleotides described herein encode two CrossMab sequences: a first immunoglobulin chain comprising a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), (iii) HCDR3 (SEQ ID NO: 24), or (iv) a combination thereof; and a second immunoglobulin chain comprising a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), (iii) LCDR3 (SEQ ID NO: 33), or (iv) a combination thereof. CH1-CLx In some embodiments, the polyribonucleotides described herein encode immunoglobulin chains of an antibody agent. CrossMab sequences are constructed using two immunoglobulin chains: a first immunoglobulin chain comprising a heavy chain variable domain comprising (i) HCDR1 (SEQ ID NO: 18), (ii) HCDR2 (SEQ ID NO: 21), and (iii) HCDR3 (SEQ ID NO: 24); and a second immunoglobulin chain comprising a light chain variable domain comprising (i) LCDR1 (SEQ ID NO: 27), (ii) LCDR2 (LAS, SEQ ID NO: 30), and (iii) LCDR3 (SEQ ID NO: 33). CH1-CLx It encodes the immunoglobulin chains of antibody agents.

[0461] In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The 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 set forth in SEQ ID NO: 36. In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLxThe antibody agent comprises a light chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The antibody agent comprises a heavy chain variable domain represented by SEQ ID NO: 36. In some embodiments, the CrossMab CH1-CLx The antibody agent comprises a light chain variable domain represented by SEQ ID NO:43.

[0462] In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The 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 set forth in SEQ ID NO: 1494. In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The antibody agent comprises a light chain variable domain having at least 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence set forth in SEQ ID NO: 43. In some embodiments, the CrossMab encoded by one or more polyribonucleotides provided herein CH1-CLx The antibody agent comprises a heavy chain variable domain represented by SEQ ID NO: 1494. In some embodiments, the CrossMab CH1-CLx The antibody agent comprises a light chain variable domain represented by SEQ ID NO:43.

[0463] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain comprising one or more mutations compared to the amino acid sequence set forth in SEQ ID NO: 1494. In some embodiments, there is a substitution mutation at residues 3 and / or 5 compared to SEQ ID NO: 1494. In some embodiments, the substitution mutation at residue 3 compared to SEQ ID NO: 1494 results in the presence of a positively charged amino acid at residue 3. Positively charged amino acids include Lys (K), Arg (R), and His (H). In some embodiments, the substitution mutation at residue 3 compared to SEQ ID NO: 1494 includes Q3H. In some embodiments, the substitution mutation at residue 5 compared to SEQ ID NO: 1494 results in the presence of a polar amino acid at residue 5. Polar amino acids include Ser (S), Thr (T), Tyr (Y), Asn (N), and Gln (Q). In some embodiments, the substitution mutation at residue 5 compared to SEQ ID NO: 1494 includes V5T.

[0464] In some embodiments, an antibody agent (e.g., a PGDM1400 antibody agent) encoded by one or more polyribonucleotides provided herein comprises a heavy chain variable domain comprising SEQ ID NO: 1494 with substitution mutations Q3H and V5T.

[0465] In some embodiments, the polyribonucleotides described herein are CH1-CLx In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of an antibody agent, the immunoglobulin chain comprising 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 set forth in SEQ ID NO: 36. CH1-CLxIn some embodiments, the polyribonucleotides described herein encode immunoglobulin chains of an antibody agent, the immunoglobulin chains comprising 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 set forth in SEQ ID NO: 43. In some embodiments, the polyribonucleotides described herein are CH1-CLx In some embodiments, the polyribonucleotides described herein encode an immunoglobulin chain of an antibody agent, the immunoglobulin chain comprising a heavy chain variable domain set forth in SEQ ID NO: 36. CH1-CLx It encodes an immunoglobulin chain of the antibody agent, and the immunoglobulin chain comprises a light chain variable domain set forth in SEQ ID NO:43.

[0466] As mentioned above, CrossMabs encoded by one or more polyribonucleotides described herein CH1-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, the CrossMab is encoded by one or more polyribonucleotides described herein. CH1-CLx The antibody agent comprises a CH3 domain that includes a G1m3, G1m17, or G1m17,1 allotype. In some embodiments, the CrossMab is encoded by one or more polyribonucleotides 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 a sequence set forth in any one of SEQ ID NOs: 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146 or 1494. In some embodiments, the CrossMab is encoded by one or more polyribonucleotides described herein. CH1-CLxThe antibody agent comprises a CH3 domain having an amino acid sequence set forth in any one of SEQ ID NOs: 113, 116, 119, 122, 125, 128, 131, 134, 137, 140, 143, 146 or 1495. In some embodiments, the CH3 domain comprises the amino acid sequence of SEQ ID NO: 1495 with substitution mutations D16E and L18M.

[0467] CrossMabs encoded by one or more polyribonucleotides described herein CH1-CLx An antibody agent can include one or more heavy chain constant domains that contain 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 described herein can be used. CH1-CLx The antibody agent may comprise an L / S mutation within the CH3 region (to enhance FcRn binding) (see Zalevsky J et al. Nat Biotechnol. 2010, which is incorporated herein by reference). Such mutations are designated M428L and N434S based on EU numbering (i.e., M88L and N94S within the CH3 domain, e.g., in SEQ ID NOs: 113, 116, or 1495), and are referred to herein as "LS" or "L / S" (see, e.g., Figure 5, panel D). In some embodiments, the CrossMab encoded by one or more polyribonucleotides described herein may be a nucleotide sequence encoding a specific nucleotide sequence. CH1-CLx The ...

Claims

1. a polyribonucleotide encoding an immunoglobulin chain of an antibody agent, the immunoglobulin chain comprising a single chain variable fragment (scFv), the scFv comprising a heavy chain variable (VH) domain, a linker, and a light chain variable (VL) domain; the VH domain comprises HCDR1, HCDR2 and HCDR3, wherein the HCDR1, the HCDR2 and the HCDR3 comprise the amino acid sequences according to SEQ ID NOs: 18, 21 and 24, respectively, and the VH domain comprises a first mutation at residue 3 and a second mutation at residue 5 compared to the amino acid sequence according to SEQ ID NO: 1494; the VL domain comprises LCDR1, LCDR2 and LCDR3, wherein the LCDR1, LCDR2 and LCDR3 comprise the amino acid sequences according to SEQ ID NOs: 27, 30 and 33, respectively; The polyribonucleotide.

2. 2. The polyribonucleotide of claim 1, wherein the first mutation comprises a substitution mutation resulting in a positively charged amino acid being present at residue 3.

3. 3. The polyribonucleotide of claim 2, wherein the positively charged amino acids comprise amino acids selected from Lys (K), Arg (R), or His (H).

4. 4. The polyribonucleotide of claim 2 or 3, wherein the positively charged amino acids include His (H) residues.

5. 5. The polyribonucleotide of claim 1, wherein the second mutation comprises a substitution mutation resulting in a polar amino acid being present at residue 5.

6. 6. The polyribonucleotide of claim 5, wherein the polar amino acids comprise amino acids selected from Ser (S), Thr (T), Tyr (Y), Asn (N), or Gln (Q).

7. 7. The polyribonucleotide of claim 5 or 6, wherein the polar amino acid comprises a Thr (T) residue.

8. The polyribonucleotide according to any one of claims 1 to 7, wherein the VH domain comprises an amino acid sequence that is at least 85% identical to the amino acid sequence according to SEQ ID NO:

36.

9. The polyribonucleotide according to any one of claims 1 to 8, wherein the VH domain comprises an amino acid sequence that is at least 90% identical to the amino acid sequence according to SEQ ID NO:

36.

10. 10. The polyribonucleotide of any one of claims 1 to 9, wherein the VH domain comprises the amino acid sequence of SEQ ID NO: 1494 with substitution mutations Q3H and V5T.

11. The polyribonucleotide according to any one of claims 1 to 10, wherein the VH domain comprises an amino acid sequence according to SEQ ID NO:

36.

12. 12. The polyribonucleotide of claim 1, wherein the VL domain comprises an amino acid sequence that is at least 85% identical to the amino acid sequence according to SEQ ID NO:

43.

13. 13. The polyribonucleotide of claim 1, wherein the VL domain comprises an amino acid sequence that is at least 90% identical to the amino acid sequence according to SEQ ID NO:

43.

14. The polyribonucleotide according to any one of claims 1 to 13, wherein the VL domain comprises an amino acid sequence according to SEQ ID NO:

43.

15. The scFv is (i) the VH domain; (ii) the linker, and (iii) the VL domain The polyribonucleotide according to any one of claims 1 to 14, comprising, in order:

16. The scFv is (i) the VL domain; (ii) the linker, and (iii) the VH domain The polyribonucleotide according to any one of claims 1 to 14, comprising, in order:

17. The polyribonucleotide according to any one of claims 1 to 16, wherein the linker comprises an amino acid sequence according to SEQ ID NO:

48.

18. The polyribonucleotide according to any one of claims 1 to 16, wherein the linker comprises an amino acid sequence according to SEQ ID NO:

59.

19. The polyribonucleotide of any one of claims 1 to 18, wherein the immunoglobulin chain further comprises a second linker following the scFv domain.

20. 20. The polyribonucleotide of claim 19, wherein the scFv and the second linker comprise or consist of an amino acid sequence according to SEQ ID NO: 64, 67, 70 or 73.

21. 21. The polyribonucleotide of claim 1, wherein the immunoglobulin chain comprises a hinge domain following the second linker.

22. 22. The polyribonucleotide of claim 21, wherein the hinge domain comprises or consists of an amino acid sequence according to SEQ ID NO:

167.

23. 10. The polyribonucleotide of claim 1, wherein the immunoglobulin chain comprises one or more constant domains, and the scFv is operably linked to the one or more constant domains.

24. The polyribonucleotide of claim 10, wherein the hinge domain is between the scFv and the one or more constant domains.

25. 12. The polyribonucleotide of claim 10 or claim 11, wherein the one or more constant domains comprise a CH3 domain.

26. 26. The polyribonucleotide of claim 25, wherein the CH3 domain comprises a G1m17,1 or G1m3 allotype.

27. 27. The polyribonucleotide of claim 26, wherein the CH3 domain comprises one or more substitution mutations, the one or more substitution mutations comprising or consisting of M88L, N94S, or a combination thereof, and the positions of the substitution mutations are compared to the amino acid sequence of SEQ ID NO: 1495.

28. 28. The polyribonucleotide of claim 27, wherein the CH3 domain comprises substitution mutations at residues 16 and 18 compared to the amino acid sequence according to SEQ ID NO: 1495.

29. 29. The polyribonucleotide of claim 28, wherein the CH3 domain comprises substitution mutations D16E and L18M compared to the amino acid sequence according to SEQ ID NO: 1495.

30. 30. The polyribonucleotide of any one of claims 25 to 29, wherein the CH3 domain comprises or consists of an amino acid sequence according to SEQ ID NO:

116.

31. 2. The polyribonucleotide of claim 1, wherein the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1343.

32. 2. The polyribonucleotide of claim 1, wherein the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1346.

33. 2. The polyribonucleotide of claim 1, wherein the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1349.

34. 2. The polyribonucleotide of claim 1, wherein the immunoglobulin chain comprises or consists of a sequence according to SEQ ID NO: 1352.

35. The polyribonucleotide according to any one of claims 1 to 34, wherein the polyribonucleotide comprises a ribonucleic acid sequence encoding a secretion signal.

36. 36. The polyribonucleotide of claim 35, wherein the secretory signal comprises a ribonucleic acid sequence that is at least 90% identical to SEQ ID NO: 4 or 8.

37. 37. The polyribonucleotide of claim 36, wherein the secretion signal comprises a ribonucleic acid sequence comprising SEQ ID NO: 4 or 8.

38. 38. The polyribonucleotide of any one of claims 1 to 37, wherein the polyribonucleotide comprises one or more non-coding sequence elements.

39. 39. The polyribonucleotide of claim 38, wherein the one or more non-coding sequence elements improve RNA stability and / or translation efficiency.

40. 40. The polyribonucleotide of claim 38 or 39, wherein the one or more non-coding sequence elements comprise a 3' untranslated region (UTR), a 5' UTR, a 5' cap, a polyadenine (poly A) tail, or a combination thereof.

41. 41. The polyribonucleotide of claim 40, wherein the polyA tail is or comprises a modified polyA sequence, preferably a stuttered polyA tail.

42. 42. The polyribonucleotide of claim 41, wherein the polyA tail comprises or consists of a sequence that is at least 90% identical to SEQ ID NO:

16.

43. The polyribonucleotide of any one of claims 40 to 42, wherein the 3'UTR comprises or consists of a nucleic acid sequence that is at least 90% identical to SEQ ID NO:

14.

44. The polyribonucleotide of any one of claims 40 to 43, wherein the 5'UTR comprises or consists of a nucleic acid sequence that is at least 90% identical to SEQ ID NO:

10.

45. The 5' cap is ２ ７，３’ -O) Gppp(m ２’－Ｏ 45. The polyribonucleotide according to any one of claims 40 to 44, wherein the polyribonucleotide is ApG.

46. 46. ​​The polyribonucleotide of any one of claims 1 to 45, wherein the polyribonucleotide comprises one or more modified ribonucleotides.

47. 47. The polyribonucleotide of claim 46, wherein the one or more modified ribonucleotides comprises pseudouridine.

48. 48. The polyribonucleotide according to any one of claims 1 to 47, wherein said polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1000.

49. 48. The polyribonucleotide according to any one of claims 1 to 47, wherein said polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1048.

50. 48. The polyribonucleotide according to any one of claims 1 to 47, wherein said polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1096.

51. 48. The polyribonucleotide according to any one of claims 1 to 47, wherein said polyribonucleotide comprises or consists of a ribonucleic acid sequence according to SEQ ID NO: 1144.

52. A polyribonucleotide comprising or consisting of a ribonucleic acid sequence according to any one of SEQ ID NOs: 1000, 1048, 1096 or 1044.

53. A composition comprising one or more polyribonucleotides according to any one of claims 1 to 52.

54. the composition further comprises a lipid nanoparticle, a polyplex (PLX), a lipidated polyplex (LPLX), or a liposome; 54. The composition of claim 53, wherein the one or more polyribonucleotides are fully or partially encapsulated in the lipid nanoparticle, the polyplex (PLX), the lipidated polyplex (LPLX), or the liposome.

55. 55. The composition of claim 53 or claim 54, wherein the composition further comprises lipid nanoparticles, and the one or more polyribonucleotides are encapsulated within the lipid nanoparticles.

56. 56. The composition of claim 54 or claim 55, wherein the lipid nanoparticles are cationic lipid nanoparticles.

57. A pharmaceutical composition comprising the composition of any one of claims 53 to 56 and at least one pharmaceutically acceptable excipient.

58. 58. The pharmaceutical composition of claim 57 for use in the treatment and / or prevention of HIV, comprising administering said pharmaceutical composition to a subject.

59. 59. A method comprising administering to a subject the pharmaceutical composition of claim 57 or claim 58.

60. 60. The method of claim 59, or the pharmaceutical composition for use of claim 58, wherein the antibody agent is expressed in the subject by administering the pharmaceutical composition to the subject.

61. In the subject, the antibody agent (a) at least 1 μg / ml in plasma; or (b) at least 1 μg / ml in serum 61. The method of claim 59 or 60, or the pharmaceutical composition for use of claim 58, wherein the antibody is expressed at a titer of

62. In the subject, the antibody agent (a) at least 10 μg / ml in plasma; or (b) at least 10 μg / ml in serum 62. The method of claim 61 , wherein the gene is expressed at a titer of

63. 63. The method of claim 62, wherein the antibody agent is detectable in the serum of the subject for a period of at least 5 days, at least 10 days, at least 15 days, at least 20 days, at least 25 days, or at least 30 days.

64. 64. The method of claim 62 or claim 63, wherein the antibody agent is detectable in the serum of the subject for a period of at least 30 days.

65. 65. The method of any one of claims 59 to 64, or the pharmaceutical composition for use of claim 58, wherein the antibody agent has the ability to neutralize one or more HIV strains when tested in a TZM-bl cell pseudoviral neutralization assay at antibody agent concentrations of up to 25 μg / ml.

66. 66. The method of any one of claims 59 to 65, or the pharmaceutical composition for use of claim 58, wherein the neutralizing activity of the antibody agent delivered as a polyribonucleotide against one or more HIV strains is greater compared to an equivalent amount of a parent control antibody delivered as a polyribonucleotide, and the parent control antibody is an IgG antibody comprising the same VH and VL domains as the antibody agent.

67. 67. The method of claim 65 or claim 66, wherein the one of more HIV strains comprises one or more HIV strains selected from ZM53M.PB12, Du156.12, Q769.d22, 0330.v4.c3, R2184.c04, 89-F1_2_25, CAP204_2_00_F6_6, Ce1176_A3, 6980.v0.c31, PVO.4, CAP45, CNE8, T250-4, 3103.v3.c10, and C1080_c3.

68. 68. The method of any one of claims 59 to 67, or the pharmaceutical composition for use of claim 58, wherein the subject has or is at risk of developing an HIV infection.

69. The method according to any one of claims 59 to 68, wherein the method is a method for treating and / or preventing HIV infection.

70. Use of a composition according to any one of claims 53 to 56 or a pharmaceutical composition according to claim 57 or claim 58 for the treatment and / or prevention of HIV in said subject.

71. 59. A method of producing an antibody agent, the method comprising administering to a cell a composition according to any one of claims 51 to 54 or a pharmaceutical composition according to claim 57 or claim 58, such that the cell expresses and secretes the antibody agent.

72. 72. The method of claim 71, wherein the cells are in the subject and the antibody agent is produced at a therapeutically relevant plasma concentration or a therapeutically relevant serum concentration.

73. 73. The method of claim 72, wherein the therapeutically relevant plasma concentration or the therapeutically relevant serum concentration is at least 10 μg / ml.

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