SARS-COV-2 spike protein binding molecule

Abstract

SARS-CoV-2 spike protein-binding molecules are disclosed. Nucleic acids and expression vectors encoding the SARS-CoV-2 spike protein-binding molecules, compositions comprising the SARS-CoV-2 spike protein-binding molecules, and methods of using the SARS-CoV-2 spike protein-binding molecules are also disclosed.

Description

[Technical Field]

[0001] This application claims priority from SG10202260528T, filed December 21, 2022, the contents and sections of which are incorporated herein by reference for all purposes.

[0002] The present disclosure relates to the field of molecular biology, and more specifically to antibody engineering. The present disclosure also relates to methods of medical treatment and prophylaxis. [Background technology]

[0003] The human infectious disease pandemic COVID-19 caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its prolonged and ongoing outbreak have resulted in devastating economic losses and loss of human life worldwide. The emergence of SARS-CoV-2 variants has presented significant challenges for the widespread treatment / prevention of COVID-19.

[0004] Antibodies that can inhibit the interaction between the spike protein of SARS-CoV-2 and SARS-CoV-2 variants and the spike protein receptor ACE2, thereby inhibiting infection of ACE2-expressing cells by such viruses, have been described, for example, in WO2022 / 245288A1 and Westendorf et al., Cell Reports (2022) 39(7):110812. However, there remains an unmet need for antibodies that can neutralize infection by a wide range of sarbecoviruses, including emerging and future SARS-CoV-2 variants, as well as other sarbecoviruses with pandemic potential. Summary of the Invention

[0005] In a first aspect, the present disclosure provides an antigen-binding molecule, optionally isolated, that binds to a sarbecovirus spike protein, comprising: (i) a VH region comprising HC-CDR1, HC-CDR2, and HC-CDR3 as set forth in column A of Table A; and (ii) a VL region comprising LC-CDR1, LC-CDR2, and LC-CDR3 as set forth in column B of Table A, wherein the sequences in columns A and B are selected from the same row of Table A.

[0006] In some embodiments, the antigen binding molecule comprises: (i) The following CDR: HC-CDR1 having the amino acid sequence of SEQ ID NO: 37 HC-CDR2 having the amino acid sequence of SEQ ID NO: 53 HC-CDR3 having the amino acid sequence of SEQ ID NO: 54 a heavy chain variable (VH) region incorporating (ii) the following CDR: LC-CDR1 having the amino acid sequence of SEQ ID NO: 60 LC-CDR2 having the amino acid sequence of SEQ ID NO: 61 LC-CDR3 having the amino acid sequence of SEQ ID NO: 62 The light chain variable (VL) region incorporating Includes.

[0007] In some embodiments, the antigen binding molecule comprises (i) a VH region comprising the amino acid sequence shown in column A of Table C, and (ii) a VL region comprising the amino acid sequence shown in column B of Table C, wherein the sequences in columns A and B are selected from the same row of Table C.

[0008] In some embodiments, the antigen binding molecule comprises: a VH region having an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 52; and A VL region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 59. Includes.

[0009] In some embodiments, the antigen binding molecule is a multispecific antigen binding molecule, and the antigen binding molecule further comprises an antigen binding domain that binds to an antigen other than a sarbecovirus spike protein.

[0010] The present disclosure also provides a chimeric antigen receptor (CAR) comprising an antigen-binding molecule according to the present disclosure. The present disclosure also provides one or more nucleic acids, optionally isolated, encoding an antigen-binding molecule according to the present disclosure or a CAR according to the present disclosure.

[0011] The present disclosure also provides one or more expression vectors comprising one or more nucleic acids according to the present disclosure. The present disclosure also provides a cell comprising an antigen-binding molecule, a CAR, one or more nucleic acids, or one or more expression vectors according to the present disclosure.

[0012] The present disclosure also provides a method comprising culturing a cell according to the present disclosure under conditions suitable for expression of the antigen binding molecule or CAR by the cell. The present disclosure also provides a composition comprising an antigen-binding molecule, a CAR, one or more nucleic acids, one or more expression vectors, or cells according to the present disclosure and a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.

[0013] In some embodiments, the composition comprises: (a) an antigen-binding molecule that binds to a sarbecovirus spike protein, comprising a VH region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 824, and a VL region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 830; or (b) an antigen-binding molecule that binds to a sarbecovirus spike protein and comprises a VH region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 839 and a VL region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 846. Further includes:

[0014] The present disclosure provides: (i) an antigen-binding molecule according to the present disclosure; and (ii) (a) an antigen-binding molecule that binds to a sarbecovirus spike protein, the antigen-binding molecule comprising a VH region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 824 and a VL region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 830; or (b) an antigen-binding molecule that binds to a sarbecovirus spike protein, the antigen-binding molecule comprising a VH region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 839 and a VL region having an amino acid sequence with at least 70% amino acid sequence identity to SEQ ID NO: 846. Also provided is a combination comprising:

[0015] The present disclosure also provides an antigen-binding molecule, a CAR, one or more nucleic acids, one or more expression vectors, a cell, a composition, or a combination according to the present disclosure for use in a method of medical treatment or prevention.

[0016] The present disclosure also provides an antigen-binding molecule, a CAR, one or more nucleic acids, one or more expression vectors, cells, compositions, or combinations according to the present disclosure for use in treating or preventing a disease or condition characterized by infection with a sarbecovirus, wherein the disease or condition characterized by infection with a sarbecovirus can be COVID-19.

[0017] The present disclosure also provides use of an antigen-binding molecule, a CAR, one or more nucleic acids, one or more expression vectors, a cell, a composition, or a combination according to the present disclosure in the manufacture of a medicament for treating or preventing a disease or condition characterized by infection by a sarbecovirus, wherein the disease or condition characterized by infection by a sarbecovirus can be COVID-19.

[0018] The present disclosure also provides a method of treating or preventing a disease or condition characterized by infection by a sarbecovirus in a subject, the method comprising administering to the subject a therapeutically or prophylactically effective amount of an antigen binding molecule, a CAR, one or more nucleic acids, one or more expression vectors, cells, compositions, or combinations according to the present disclosure in the manufacture of a medicament for treating or preventing a disease or condition characterized by infection by a sarbecovirus, wherein the disease or condition characterized by infection by a sarbecovirus can be COVID-19.

[0019] The present disclosure also provides an in vitro complex, optionally isolated, comprising an antigen-binding molecule according to the present disclosure bound to a sarbecovirus or a sarbecovirus spike protein.

[0020] The present disclosure also provides a method for detecting a sarbecovirus or a sarbecovirus spike protein in a sample, the method comprising the steps of contacting a sample containing or suspected of containing a sarbecovirus or a sarbecovirus spike protein with an antigen-binding molecule according to the present disclosure, and detecting formation of a complex between the antigen-binding molecule and the sarbecovirus or the sarbecovirus spike protein.

[0021] The present disclosure also provides a method for selecting or stratifying a subject for treatment with a drug that targets a sarbecovirus, the method comprising the steps of contacting in vitro a sample from the subject with an antigen-binding molecule according to the present disclosure, and detecting formation of a complex between the antigen-binding molecule and the sarbecovirus or a sarbecovirus spike protein.

[0022] The present disclosure also provides the use of an antigen-binding molecule according to the present disclosure as a diagnostic or prognostic agent in vitro or in vivo. [Brief explanation of the drawings]

[0023] [Figure 1] Figure 1A is a graph of the % inhibition of infection of ACE2-expressing cells by pseudoviruses expressing the spike protein of SARS-CoV-2 (Figure 1B) BA.2.75.2, (Figure 1C) BF.7, (Figure 1D) BA.4.6.1, (Figure 1E) BQ.1.1, and (Figure 1F) XBB.1 for the indicated antibodies, as determined by pseudovirus neutralization test (pVNT). [Figure 2] Figure 2A is a table summarizing the IC50 values, as determined by pVNT, for the indicated antibodies / antibody combinations for inhibition of infection of ACE2-expressing cells by pseudoviruses expressing the indicated sarbecovirus spike proteins. Figure 2B is a table summarizing the IC50 values, as determined by pVNT, for the indicated antibodies / antibody combinations for inhibition of infection of ACE2-expressing cells by pseudoviruses expressing the indicated sarbecovirus spike proteins. Figure 2C is a table summarizing the IC50 values, as determined by pVNT, for the indicated antibodies / antibody combinations for inhibition of infection of ACE2-expressing cells by pseudoviruses expressing the indicated sarbecovirus spike proteins. [Figure 3] 1 is a bar graph showing IC50 values ​​for inhibition of infection of ACE2-expressing cells by pseudoviruses expressing the spike proteins of the indicated sarbecoviruses by the indicated antibodies, as determined by pVNT. [Figure 4] 1 is a bar graph showing IC50 values ​​for inhibition of infection of ACE2-expressing cells by pseudoviruses expressing the spike proteins of the indicated sarbecoviruses by the indicated antibodies / antibody combinations as determined by pVNT performed in the presence of human serum. [Figure 5] Graph showing IC50 values ​​for inhibition of the interaction between human ACE2 and the RBD of the spike proteins of the indicated sarbecoviruses by the indicated antibodies / antibody combinations as determined by surrogate virus neutralization test (sVNT). DETAILED DESCRIPTION OF THE INVENTION

[0024] explanation The present disclosure provides antigen-binding molecules that bind to sarbecovirus spike proteins (e.g., SARS-CoV-2 spike proteins and / or SARS-CoV-2 variant spike proteins) and have novel biophysical and / or functional properties compared to antigen-binding molecules disclosed in the prior art.

[0025] In particular, the present disclosure provides antigen binding molecules that bind to sarbecovirus spike proteins and inhibit the interaction between the sarbecovirus spike proteins and ACE2, which are useful for inhibiting infection of ACE2-expressing cells by sarbecoviruses containing such spike proteins.

[0026] The antigen binding molecules of the present disclosure are capable of interacting between ACE2 and a wide variety of sarbecovirus spike proteins, including spike proteins from a wide range of SARS-CoV-2 variants. Thus, the antigen binding molecules of the present disclosure are useful for treating and preventing diseases caused by a wide range of sarbecoviruses, including a wide range of SARS-CoV-2 variants. It is also demonstrated herein that the antigen binding molecules of the present disclosure inhibit the interaction between ACE2 and sarbecovirus spike proteins with increased potency compared to known sarbecovirus spike protein-binding antibodies.

[0027] The present disclosure also provides compositions comprising and therapeutic / prophylactic interventions using novel combinations of antigen binding molecules that provide increased potency in inhibiting infection of ACE2-expressing cells by sarbecoviruses and / or inhibiting infection of ACE2-expressing cells by a broad range of sarbecoviruses compared to known compositions / interventions.

[0028] Sarbecoviruses, SARSr-CoV, SARS-CoV-2 and SARS-CoV-2 variants The present disclosure relates to sarbecoviruses, which are members of the Sarbecovirus subgenus of coronaviruses in the Betacoronavirus genus that infect humans, bats, and certain other mammals. They are enveloped, positive-sense, single-stranded RNA viruses.

[0029] Sarbecoviruses are divided into three major clades based on their evolutionary relationships: clades 1, 2, and 3; see, e.g., Xiang et al., Cell Rep. (2022) 39(13):111004 and Tortorici et al., Nature (2021) 597:103-108.

[0030] Sarbecoviruses in Clade 1 can be further divided into Clades 1a, 1b, and 1c. Clade 1a sarbecoviruses include SARS-CoV (also known as SARS-CoV-1), WIV-1, LYRa11, Rs4231, BtSY1, RsSHC014, and Rs9401. Clade 1b sarbecoviruses include SARS-CoV-2, SARS-CoV-2 variants, RaTG13, BANAL-20-51, BANAL-20-52, BANAL-20-236, BANAL-20-103, Rc-o319, RsSTT182, BtSY2, GX-P5L, and GD-1. Clade 1c sarbecoviruses include RaTG15 and RpYN04. Clade 2 sarbecoviruses include RmYN02, RacCS203, SL-ZX45, SL-ZXC21, BANAL-20-116, BANAL-20-247, PrC31, RpYN06, Rm1, Rf1, Rp3, HKU3-1, JTMC15, SX2013, HeB2013, Rs4237, 16BO133, and Anlong-103. Clade 3 sarbecoviruses include BtKY72, BM48-31, and Khosta-2.

[0031] In some embodiments, the sarbecovirus according to the present disclosure is a Clade 1, Clade 2, or Clade 3 sarbecovirus. In some embodiments, the sarbecovirus is a Clade 1 or Clade 3 sarbecovirus. In some embodiments, the sarbecovirus is a Clade 1b or Clade 3 sarbecovirus. In some embodiments, the sarbecovirus is not a Clade 2 sarbecovirus. In some embodiments, the sarbecovirus is a Clade 1 sarbecovirus. In some embodiments, the sarbecovirus is a Clade 1a or 1b sarbecovirus. In some embodiments, the sarbecovirus is a Clade 1b sarbecovirus.

[0032] In some embodiments, a sarbecovirus according to the present disclosure can be a sarbecovirus having a nucleotide sequence with at least 60% (e.g., one of ≧60%, ≧65%, ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) sequence identity to the nucleotide sequence of GenBank: MN908947.3.

[0033] In some embodiments, the sarbecovirus according to the present disclosure is severe acute respiratory syndrome-associated coronavirus (SARSr-CoV). The virology of SARSr-CoV and the epidemiology of disease associated with SARSr-CoV infection are reviewed, for example, in Cheng et al., Clin Microbiol Rev (2007) 20(4):660-694 and de Wit et al., Nat Rev Microbiol (2016) 14:523-534, both of which are incorporated herein by reference in their entireties.

[0034] Two strains of SARSr-CoV have caused severe outbreaks of severe respiratory disease in humans: SARS-CoV caused the severe acute respiratory syndrome (SARS) outbreak between 2002 and 2003, and SARS-CoV-2 caused the coronavirus disease 2019 (COVID-19) pandemic. There are hundreds of strains of SARSr-CoV known to infect exclusively nonhuman species; bats are the primary reservoir for many strains of SARS-related coronaviruses.

[0035] In some embodiments, the sarbecovirus according to the present disclosure is SARS-CoV-2 or a SARS-CoV-2 variant. As used herein, "SARS-CoV-2" refers to SARSr-CoV having the nucleotide sequence of GenBank: MN908947.3 ("Severe acute respiratory syndrome coronavirus 2 isolate Wuhan-Hu-1, complete genome") reported in Wu et al., Nature (2020) 579:265-269.

[0036] Numerous SARS-CoV-2 variants have been observed and are described, for example, in Planas et al., Nat. Comm. (2023) 14:824, Habib et al., Microbiol Resour Announc. (2023) 12(3):e00001-23, Katzmarzyk et al., Front Immunol. (2023) 14:1288794, Lasrado et al., Vaccine (2023) 41(47):6904-6909 and Rahman et al., Microbiol Resour Announc. (2023) 12(10):e00562-23.

[0037] "SARSr-CoV" according to the present disclosure may refer to a sarbecovirus having a nucleotide sequence with at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the nucleotide sequence of GenBank: MN908947.3.

[0038] As used herein, "SARS-CoV-2 variant" refers to a SARSr-CoV having a nucleotide sequence with at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) sequence identity to the nucleotide sequence of GenBank:MN908947.3, where the nucleotide sequence is not identical to the nucleotide sequence of GenBank:MN908947.3.

[0039] SARS-CoV-2 variants of particular interest in relation to the present disclosure include: BA.1 (also known as Omicron; B.1.1.529; e.g., represented by GISAID accession EPI_ISL_7358094.2); Omicron subvariants, such as BA.2 (e.g., represented by GISAID accession EPI_ISL_6795834.2), BA.5 (GISAID accession EPI_ISL_12268495.2), BA.2.75 (e.g., GISAID accession EPI_ISL_12268495.2), BA.2.75 (e.g., GISAID accession EPI_ISL_12268495.2), BA.3 (e.g., GISAID accession EPI_ISL_12268495.2), BA.4 (e.g., GISAID accession EPI_ISL_12268495.2), BA.5 (e.g., GISAID accession EPI_ISL_12268495.2), BA.4.75 (e.g., GISAID accession EPI_ISL_12268495.2), BA. ... SL_13692860), BA.2.75.2 (e.g., represented by GISAID accession EPI_ISL_15731524), BA.4.6.1 (e.g., represented by GISAID accession EPI_ISL_13925521), BF.7 (e.g., represented by GISAID accession EPI_ISL_13972569), BQ.1.1 (e.g., represented by GISAID accession EPI_ISL_15731523), XBB (e.g., GISAI XBB.1 (e.g., represented by GISAID accession EPI_ISL_15503011); XBB.1.16 (e.g., represented by GISAID accession EPI_ISL_17646715); XBB.2.3 (e.g., represented by GISAID accession EPI_ISL_17719186); EG.5 (e.g., represented by EPI_ISL_17976635), EG.5.1 (e.g., GISAID These include B.1.1.7 (also known as alpha; GISAID accession EPI_ISL_674612); B.1.351 (also known as beta and 501Y.V2; GISAID accession EPI_ISL_940877); B.1.617.2 (also known as delta; GISAID accession EPI_ISL_1921353); and P.1 (also known as gamma; GISAID accession EPI_ISL_2777382).

[0040] Thus, in some embodiments, the SARS-CoV-2 variant according to the present disclosure is selected from BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, XBB, XBB.1, XBB.1.16, XBB.2.3, EG.5, EG.5.1, B.1.1.7, B.1.351, B.1.617.2 and P.1.

[0041] SARS-CoV-2 spike protein and SARS-CoV-2 variant spike protein The sarbecovirus genome encodes four major structural proteins: spike (S) protein, envelope (E) protein, membrane (M) protein, and nucleocapsid (N) protein. The present disclosure particularly relates to antigen-binding molecules that bind to the spike protein of sarbecoviruses.

[0042] The canonical spike protein of SARS-CoV-2 (i.e., the spike protein encoded by the nucleotide sequence of GenBank: MN908947.3) has the amino acid sequence shown in SEQ ID NO: 1. The SARS-CoV-2 spike protein comprises an S1 (SEQ ID NO: 6) and an S2 (SEQ ID NO: 9) subunit. The S1 subunit contains the minimal receptor binding domain (RBD; SEQ ID NO: 7) through which SARS-CoV-2 binds to ACE2 expressed by host cells. The RBD in turn contains a receptor binding motif (RBM; SEQ ID NO: 8), which is the region of the RBM that contacts ACE2.

[0043] As used herein, "SARS-CoV-2 spike protein" refers to a polypeptide having the amino acid sequence of SEQ ID NO: 1. The RBD of the SARS-CoV-2 spike protein refers to the amino acid sequence of SEQ ID NO: 7. The RBM of the SARS-CoV-2 spike protein refers to the amino acid sequence of SEQ ID NO: 8.

[0044] A number of variants of the SARS-CoV-2 spike protein (i.e., encoded by SARS-CoV-2 variants) have been reported that contain one or more amino acid substitutions, deletions, or insertions in the amino acid sequence of the spike protein. Such proteins may be referred to herein as SARS-CoV-2 variant spike proteins.

[0045] A "sarbecovirus spike protein" according to the present disclosure refers to a polypeptide having an amino acid sequence with at least 60% (e.g., one of ≧60%, ≧65%, ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) amino acid sequence identity to SEQ ID NO:1. A "SARSr-CoV spike protein" according to the present disclosure refers to a polypeptide having an amino acid sequence with at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO:1.

[0046] A "SARS-CoV-2 variant spike protein" according to the present disclosure refers to a polypeptide having an amino acid sequence with at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO:1, where the amino acid sequence is not identical to SEQ ID NO:1.

[0047] In some embodiments, the SARS-CoV-2 variant spike protein comprises an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO:7, wherein the amino acid sequence is not identical to SEQ ID NO:7. In some embodiments, the SARS-CoV-2 variant spike protein comprises an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO:8, wherein the amino acid sequence is not identical to SEQ ID NO:8.

[0048] The following table summarizes the differences (i.e., amino acid substitutions and deletions (Δ) in the amino acid sequences in the spike protein encoded by the SARS-CoV-2 variants of particular interest). The SARS-CoV-2 variant spike protein differences shown in Table 1 are obtained from pandemic information (Gangavarapu et al., Nature Methods (2023) 20:512-522). The numbering of SARS-CoV-2 spike protein residues and variant positions can be determined by comparison with SEQ ID NO: 1 of the present disclosure.

[0049] [Table 1-1]

[0050] [Table 1-2]

[0051] [Table 1-3]

[0052] In some embodiments, a SARS-CoV-2 variant spike protein according to the present disclosure has an amino acid sequence with at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 and contains one or more differences as shown in Table 1 above.

[0053] In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 and includes the difference(s) set forth in rows 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 of column A of Table 1 above. By way of example, in some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 and includes N440K, S477N, T478K, E484A, Q493R, Q498R, N501Y, Y505H, G446S, and G496S (i.e., the RBM differences of BA.1, as set forth in row 1).

[0054] In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 and includes the difference(s) set forth in rows 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 of column B of Table 1 above. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 and includes the difference(s) set forth in rows 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, or 17 of column C of Table 1 above.

[0055] In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, and includes the difference(s) set forth in rows 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 of columns A and B of Table 1 above. By way of example, in some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1 and comprises N440K, S477N, T478K, E484A, Q493R, Q498R, N501Y, Y505H, G446S, G496S, G339D, S373P, S375F, K417N and S371L (i.e., the RBD differences of BA.1, as shown in row 1).

[0056] In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, and includes the difference(s) set forth in rows 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 of columns A, B, and C of Table 1 above. By way of example, in some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% sequence identity to the amino acid sequence of SEQ ID NO: 1, and includes the difference(s) set forth in rows 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or 18 of columns A, B, and C of Table 1 above. F, including K417N, S371L, A67V, Δ69, Δ70, T95I, G142D, Δ143, Δ144, Δ145, Δ211, L212I, +214EPE, T547K, D614G, H655Y, N679K, P681H, N764K, D796Y, N856K, Q954H, N969K, and L981F (i.e., spike protein differences in BA.1, as shown in row 1).

[0057] In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of the amino acid sequence of a spike protein encoded by a SARS-CoV-2 variant selected from BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, XBB, XBB.1, XBB.1.16, XBB.2.3, EG.5, EG.5.1, B.1.1.7, B.1.351, B.1.617.2, and P.1.

[0058] In some embodiments, a SARS-CoV-2 variant spike protein according to the present disclosure comprises an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO:703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718 or 719.

[0059] In some embodiments, a SARS-CoV-2 variant spike protein according to the present disclosure comprises an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 686, 687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 699, 700, 701 or 702.

[0060] In some embodiments, a SARS-CoV-2 variant spike protein according to the present disclosure comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27.

[0061] In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 11, and includes the difference(s) set forth in (i) row 1 of column A of Table 1; (ii) row 1 of columns A and B of Table 1; or (iii) row 1 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 12, and includes the difference(s) set forth in (i) row 2 of column A of Table 1; (ii) row 2 of columns A and B of Table 1; or (iii) row 2 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 13, and includes the difference(s) set forth in (i) row 3 of column A of Table 1; (ii) row 3 of columns A and B of Table 1; or (iii) row 3 of columns A, B, and C of Table 1.In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 14, and includes the difference(s) set forth in (i) row 4 of column A of Table 1; (ii) row 4 of columns A and B of Table 1; or (iii) row 4 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 15, and includes the difference(s) set forth in (i) row 5 of column A of Table 1; (ii) row 5 of columns A and B of Table 1; or (iii) row 5 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 16, and includes the difference(s) set forth in (i) row 6 of column A of Table 1; (ii) row 6 of columns A and B of Table 1; or (iii) row 6 of columns A, B, and C of Table 1.In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 17, and includes the difference(s) set forth in (i) row 7 of column A of Table 1; (ii) row 7 of columns A and B of Table 1; or (iii) row 7 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 18, and includes the difference(s) set forth in (i) row 8 of column A of Table 1; (ii) row 8 of columns A and B of Table 1; or (iii) row 8 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 19, and includes the difference(s) set forth in (i) row 9 of column A of Table 1; (ii) row 9 of columns A and B of Table 1; or (iii) row 9 of columns A, B, and C of Table 1.In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO:20, and includes the difference(s) set forth in (i) row 10 of column A of Table 1; (ii) row 10 of columns A and B of Table 1; or (iii) row 10 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO:21, and includes the difference(s) set forth in (i) row 11 of column A of Table 1; (ii) row 11 of columns A and B of Table 1; or (iii) row 11 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO:22, and includes the difference(s) set forth in (i) row 12 of column A of Table 1; (ii) row 12 of columns A and B of Table 1; or (iii) row 12 of columns A, B, and C of Table 1.In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO:23, and contains the difference(s) set forth in (i) row 13 of column A of Table 1; (ii) row 13 of columns A and B of Table 1; or (iii) row 13 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) sequence identity to the amino acid sequence of SEQ ID NO:24, and includes the difference(s) set forth in (i) row 14 of column A of Table 1; (ii) row 14 of columns A and B of Table 1; or (iii) row 14 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO:25, and contains the difference(s) set forth in (i) row 15 of column A of Table 1; (ii) row 15 of columns A and B of Table 1; or (iii) row 15 of columns A, B, and C of Table 1.In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO:26, and includes the difference(s) set forth in (i) row 16 of column A of Table 1; (ii) row 15 of columns A and B of Table 1; or (iii) row 16 of columns A, B, and C of Table 1. In some embodiments, the SARS-CoV-2 variant spike protein comprises or consists of an amino acid sequence having at least 70% (e.g., one of: ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO:27, and contains the difference(s) set forth in: (i) row 17 of column A of Table 1; (ii) row 17 of columns A and B of Table 1; or (iii) row 15 of columns A, B, and C of Table 1.

[0062] ACE2 Angiotensin-converting enzyme 2 (ACE2) is the cellular entry point for SARS-CoV through interaction with the spike protein, which binds to the extracellular domain of ACE2 (Zhou et al., Nature (2020) 579:270-273; Hoffmann et al., Cell (2020) 181:271-280).

[0063] ACE2 is a type I single-pass transmembrane carboxypeptidase that is attached to the cell membrane of cells in the outer surface tissues of the lungs, arteries, heart, kidneys, and intestinal tract. The structure and function of ACE2 are described, for example, in Hamming et al., J Pathol (2004) 203(2):631-637, the entire contents of which are incorporated herein by reference.

[0064] As used herein, "ACE2" refers to ACE2 from any species, including ACE2 isoforms, fragments, variants, or homologs from any species. In some embodiments, the ACE2 is from a mammal (e.g., therian, placental, epizootic, preptotheria, archontan, primate (rhesus monkey, cynomolgus monkey, non-human primate, or human)). In some embodiments, the ACE2 is from a human, bat, pangolin, civet, or pig. Optionally, an isoform, fragment, variant or homologue of ACE2 may be characterized as having at least 70% (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99% or 100%) sequence identity to the amino acid sequence of an immature or mature ACE2 isoform from a given species, e.g., human.

[0065] Human ACE2 isoform 1 is shown in SEQ ID NO: 28, and human ACE2 isoform 2 is shown in SEQ ID NO: 35. The extracellular domain of human ACE2 is shown in SEQ ID NO: 30.

[0066] A fragment of ACE2 can have a minimum length of one of 25, 50, 100, 200, 300, 400, 500, 600, 700, or 800 amino acids, and a maximum length of one of 50, 100, 200, 300, 400, 500, 600, 700, or 800 amino acids. A fragment of ACE2 can, for example, exhibit association with a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins).

[0067] In some embodiments, ACE2 comprises or consists of an amino acid sequence having at least 70%, preferably 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 28 or 35.

[0068] In some embodiments, the fragment of ACE2 comprises or consists of an amino acid sequence having at least 70%, preferably at least one of 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity to SEQ ID NO: 30.

[0069] Antigen-binding molecules of the present disclosure The present disclosure provides antigen-binding molecules capable of binding to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). Such antigen-binding molecules may also be described as antigen-binding molecules that bind to related proteins.

[0070] An "antigen-binding molecule" refers to a molecule that binds to a given target antigen. Antigen-binding molecules include antibodies (i.e., immunoglobulins (Ig)) and antigen-binding fragments thereof. As used herein, "antibody" includes monoclonal antibodies, polyclonal antibodies, monospecific and multispecific (e.g., bispecific, trispecific, etc.) antibodies, as well as antibody-derived antigen-binding molecules such as scFv, scFab, diabodies, triabodies, scFv-Fc, minibodies, single-domain antibodies (e.g., VhH), etc. Antigen-binding fragments of antibodies include, for example, Fv, Fab, F(ab')2, and F(ab')2 fragments. In some embodiments, the antigen-binding molecule may be an antibody or an antigen-binding fragment thereof.

[0071] Antigen-binding molecules according to the present disclosure also include antibody-derived molecules, e.g., molecules comprising an antigen-binding region / domain derived from an antibody. Antibody-derived antigen-binding molecules may comprise an antigen-binding region / domain that comprises or consists of the antigen-binding region of an antibody (e.g., an antigen-binding fragment of an antibody). In some embodiments, the antigen-binding region / domain of an antibody-derived antigen-binding molecule may be or comprise the Fv (e.g., provided as an scFv) or Fab region of an antibody, or a whole antibody. For example, antigen-binding molecules according to the present disclosure include antibody-drug conjugates (ADCs) comprising a (cytotoxic) drug moiety (e.g., as described herein below). Antigen-binding molecules according to the present disclosure also include immune cell engager molecules containing a domain (effector) for recruiting immune cells, including multispecific antigen-binding molecules such as BiTE, BiKE, and TriKE (reviewed, for example, in Goebeler and Bargou, Nat. Rev. Clin. Oncol. (2020) 17:418-434 and Ellerman, Methods (2019) 154:102-117, both of which are incorporated herein by reference in their entireties). Antigen-binding molecules according to the present disclosure also include chimeric antigen receptors (CARs), which are recombinant receptors that provide both antigen binding and T cell activation functions (CAR structure, function, and operation are reviewed, for example, in Dotti et al., Immunol Rev (2014) 257(1) and Jayaraman et al., EBioMedicine (2020) 58:102-931, both of which are incorporated herein by reference in their entireties).

[0072] The antigen-binding molecules of the present disclosure comprise one or more moieties capable of binding to a target antigen(s). In some embodiments, the moiety capable of binding to a target antigen comprises an antibody heavy chain variable region (VH) and an antibody light chain variable region (VL) of an antibody capable of specifically binding to the target antigen. In some embodiments, the moiety capable of binding to a target antigen comprises or consists of an aptamer, e.g., a nucleic acid aptamer, capable of binding to the target antigen (e.g., as reviewed in Zhou and Rossi Nat Rev Drug Discov. 2017 16(3):181-202). In some embodiments, the moiety capable of binding to a target antigen comprises or consists of an antigen-binding peptide / polypeptide, such as a peptide aptamer, thioredoxin, monobody, anticalin, Kunitz domain, avimer, knottin, fynomer, atrimer, DARPin, affibody, nanobody (i.e., single domain antibody (sdAb)), affilin, armadillo repeat protein (ArmRP), OBody, or fibronectin—reviewed, for example, in Reverdatto et al., Curr Top Med Chem. 2015;15(12):1082-1101, which is incorporated herein by reference in its entirety (see also, for example, Boersma et al., J Biol Chem (2011) 286:41273-85 and Emanuel et al., Mabs (2011) 3:38-48).

[0073] As used herein, "peptide" refers to a chain of two or more amino acid monomers linked by peptide bonds. Typically, peptides have a length in the range of about 2 to 50 amino acids. A "polypeptide" is a polymeric chain of two or more peptides. Typically, a polypeptide has a length greater than about 50 amino acids.

[0074] The antigen-binding molecule of the present disclosure generally comprises an antigen-binding domain comprising VH and VL of an antibody capable of specifically binding to a target antigen. The antigen-binding domain formed by VH and VL may also be referred to herein as an Fv region.

[0075] An antigen-binding molecule may be or may comprise an antigen-binding polypeptide or an antigen-binding polypeptide complex. An antigen-binding molecule may comprise more than one polypeptide that together form an antigen-binding domain. The polypeptides may be covalently or non-covalently bound. In some embodiments, the polypeptide forms part of a larger polypeptide that comprises the polypeptide (e.g., in the case of an scFv comprising a VH and a VL, or in the case of an scFab comprising a VH-CH1 and a VL-CL).

[0076] An antigen-binding molecule can refer to a non-covalent or covalent complex of more than one polypeptide (e.g., 2, 3, 4, 6, or 8 polypeptides), for example, an IgG-like antigen-binding molecule comprising two heavy chain polypeptides and two light chain polypeptides.

[0077] The antigen-binding molecules of the present disclosure can be designed and prepared using the sequence of monoclonal antibodies (mAbs). Antigen-binding regions of antibodies, such as single-chain variable fragments (scFv), Fab and F(ab')2 fragments, can also be used / provided. An "antigen-binding region" is any fragment of an antibody that binds to the target for which the given antibody is specific.

[0078] Antibodies generally contain six complementarity determining regions (CDRs): three in the heavy chain variable (VH) region: HC-CDR1, HC-CDR2, and HC-CDR3, and three in the light chain variable (VL) region: LC-CDR1, LC-CDR2, and LC-CDR3. Together, the six CDRs determine the paratope of the antibody, which is the portion of the antibody that binds to the target antigen.

[0079] The VH and VL regions comprise framework regions (FRs) on either side of each CDR, which provide a scaffold for the CDRs. From N- to C-terminus, the VH region comprises the following structure: N-terminus-[HC-FR1]-[HC-CDR1]-[HC-FR2]-[HC-CDR2]-[HC-FR3]-[HC-CDR3]-[HC-FR4]-C-terminus, and the VL region comprises the following structure: N-terminus-[LC-FR1]-[LC-CDR1]-[LC-FR2]-[LC-CDR2]-[LC-FR3]-[LC-CDR3]-[LC-FR4]-C-terminus.

[0080] There are several different conventions for determining the CDRs and FRs of antibodies, such as those described in Kabat et al., Sequences of Proteins of Immunological Interest, 5th Ed. Public Health Service, National Institutes of Health, Bethesda, MD (1991), Chothia et al., J. Mol. Biol. 196:901-917 (1987), and VBASE2, described in Retter et al., Nucl. Acids Res. (2005) 33(suppl 1):D671-D674. The CDRs and FRs of the VH and VL regions of the antibody clones described herein were determined according to the international IMGT (ImMunoGeneTics) information system (LeFranc et al., Nucleic Acids Res. (2015) 43 (Database issue): D413-22) using the IMGT V-DOMAIN numbering rules described in Lefranc et al., Dev. Comp. Immunol. (2003) 27: 55-77. In a preferred embodiment, the CDRs and FRs of the antigen-binding molecules referred to herein are determined according to the IMGT information system.

[0081] In some embodiments, the antigen binding molecule comprises the CDRs of an antigen binding molecule that binds to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). In some embodiments, the antigen binding molecule comprises the FRs of an antigen binding molecule that binds to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). In some embodiments, the antigen binding molecule comprises the CDRs and FRs of an antigen binding molecule that binds to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). That is, in some embodiments, the antigen binding molecule comprises the VH and VL regions of an antigen binding molecule that binds to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins).

[0082] In some embodiments, the antigen-binding molecule comprises the CDR, FR and / or VH and / or VL regions of an antibody described herein (e.g., an antibody in Table C herein) or the CDR, FR and / or VH and / or VL regions derived from those of an antibody described herein (e.g., an antibody in Table C herein).

[0083] In some embodiments, the antigen binding molecule comprises: HC-CDR1 (or 1 or 2 or 3 amino acids in HC-CDR1) as shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column A of Table A HC-CDR1 (or a variant thereof in which one, two or three amino acids in HC-CDR2 are replaced with another amino acid), HC-CDR2 (or a variant thereof in which one, two or three amino acids in HC-CDR2 are replaced with another amino acid), and HC-CDR3 (or a variant thereof in which one, two or three amino acids in HC-CDR3 are replaced with another amino acid), wherein the HC-CDR1, HC-CDR2 and HC-CDR3 sequences in column A are selected from the same row of Table A. Contains the VH region.

[0084] By way of example, in some embodiments, the antigen-binding molecule comprises a VH region comprising an HC-CDR1 having the amino acid sequence of SEQ ID NO: 37 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 37 are substituted with another amino acid), an HC-CDR2 having the amino acid sequence of SEQ ID NO: 38 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 38 are substituted with another amino acid), and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 39 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 39 are substituted with another amino acid). It will be understood that the HC-CDR1, HC-CDR2, and HC-CDR3 sequences in the foregoing sentence are selected from the same row (row 1) of column A of Table A.

[0085] In some embodiments, the antigen binding molecule comprises: HC-FR1 (or a variant thereof in which one or two or three amino acids in HC-FR1 are substituted with other amino acids), HC-FR2 (or a variant thereof in which one or two or three amino acids in HC-FR1 are substituted with other amino acids), or or a variant thereof in which one, two or three amino acids in HC-FR2 are substituted with another amino acid), HC-FR3 (or a variant thereof in which one, two or three amino acids in HC-FR3 are substituted with another amino acid), and HC-FR4 (or a variant thereof in which one, two or three amino acids in HC-FR4 are substituted with another amino acid), wherein the HC-FR1, HC-FR2, HC-FR3 and HC-FR4 sequences in column A are selected from the same row of Table B; Contains the VH region.

[0086] By way of example, in some embodiments, the antigen-binding molecule comprises a VH region comprising HC-FR1 having the amino acid sequence of SEQ ID NO: 40 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 40 are substituted with other amino acids), HC-FR2 having the amino acid sequence of SEQ ID NO: 41 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 41 are substituted with other amino acids), HC-FR3 having the amino acid sequence of SEQ ID NO: 42 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 42 are substituted with other amino acids), and HC-FR4 having the amino acid sequence of SEQ ID NO: 43 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 43 are substituted with other amino acids). It will be understood that the HC-FR1, HC-FR2, HC-FR3, and HC-FR4 sequences in the foregoing sentence are selected from the same row (row 1) of column A of Table B.

[0087] In some embodiments, the antigen binding molecule comprises: HC-CDR1 (or a CDR thereof in which one or two or three amino acids in the HC-CDR1 are substituted with another amino acid) as set forth in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column A of Table A HC-CDR1, HC-CDR2 and HC-CDR3 (or a variant thereof in which one, two or three amino acids in HC-CDR2 are replaced with another amino acid), HC-CDR2 (or a variant thereof in which one, two or three amino acids in HC-CDR2 are replaced with another amino acid), and HC-CDR3 (or a variant thereof in which one, two or three amino acids in HC-CDR3 are replaced with another amino acid), wherein the HC-CDR1, HC-CDR2 and HC-CDR3 sequences in column A are selected from the same row of Table A; and HC-FR1 (or a variant thereof in which one, two or three amino acids in HC-FR1 are substituted with another amino acid), HC-FR2 (or one or two or three amino acids in HC-FR2) shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column A of Table B is a variant thereof in which two or three amino acids in HC-FR3 are substituted with other amino acids), HC-FR3 (or a variant thereof in which one, two or three amino acids in HC-FR3 are substituted with other amino acids), and HC-FR4 (or a variant thereof in which one, two or three amino acids in HC-FR4 are substituted with other amino acids), wherein the HC-FR1, HC-FR2, HC-FR3 and HC-FR4 sequences in column A are selected from the same row of Table B. HC-FR1, HC-FR2, HC-FR3 and HC-FR4 The VH region comprises:

[0088] In some embodiments, the antigen binding molecule comprises: HC-CDR1 (or HC - HC-CDR1 (or a variant thereof in which 1, 2 or 3 amino acids in HC-CDR2 are replaced by another amino acid), HC-CDR2 (or a variant thereof in which 1, 2 or 3 amino acids in HC-CDR2 are replaced by another amino acid) and HC-CDR3 (or a variant thereof in which 1, 2 or 3 amino acids in HC-CDR3 are replaced by another amino acid); and HC-FR1 (or 1 or 2 or 3 amino acids in HC-FR1) shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column A of Table B HC-FR2 (or a variant thereof in which one, two, or three amino acids in HC-FR2 are substituted with another amino acid), HC-FR3 (or a variant thereof in which one, two, or three amino acids in HC-FR3 are substituted with another amino acid), and HC-FR4 (or a variant thereof in which one, two, or three amino acids in HC-FR4 are substituted with another amino acid). Includes wherein the HC-CDR1, HC-CDR2, HC-CDR3 sequences in column A of Table A and the HC-FR1, HC-FR2, HC-FR3 and HC-FR4 sequences in column B of Table B are selected from rows having the same number, Contains the VH region.

[0089] By way of example, in some embodiments, the antigen-binding molecule comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 37 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 37 are substituted with other amino acids), an HC-CDR2 having the amino acid sequence of SEQ ID NO: 38 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 38 are substituted with other amino acids), and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 39 (or a variant thereof in which one, two, or three amino acids in SEQ ID NO: 39 are substituted with other amino acids), an HC-FR1 having the amino acid sequence of SEQ ID NO: 40 (or and HC-FR4 having the amino acid sequence of SEQ ID NO: 43 (or a variant thereof in which one, two or three amino acids in SEQ ID NO: 43 are substituted with another amino acid). It will be understood that the HC-CDR1, HC-CDR2 and HC-CDR3 sequences in the foregoing sentence are selected from row 1 of column A of Table A, and the HC-FR1, HC-FR2, HC-FR3 and HC-FR4 sequences are selected from the row having the same number (row 1) of column A of Table B.

[0090] In some embodiments, the antigen binding molecule is selected from the group consisting of: The VH region has at least 70%, preferably ≥80%, ≥85%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100% amino acid sequence identity to the amino acid sequence of a VH region sequence selected from 46, 47, 48, 49, 50 or 51.

[0091] In some embodiments, the antigen binding molecule comprises: LC-CDR1 (or 1 or 2 or 3 amino acids in LC-CDR1) shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column B of Table A LC-CDR1 (or a variant thereof in which one, two or three amino acids in LC-CDR2 are substituted with another amino acid), LC-CDR2 (or a variant thereof in which one, two or three amino acids in LC-CDR2 are substituted with another amino acid), and LC-CDR3 (or a variant thereof in which one, two or three amino acids in LC-CDR3 are substituted with another amino acid), wherein the LC-CDR1, LC-CDR2 and LC-CDR3 sequences in column B are selected from the same row of Table A. Contains the VL region.

[0092] In some embodiments, the antigen binding molecule comprises: LC-FR1 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR2 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR3 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR4 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR5 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR6 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR7 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR8 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR9 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR10 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR11 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR12 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR13 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR14 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR15 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids), LC-FR16 (or a variant or a variant thereof in which one, two, or three amino acids in LC-FR2 are substituted with another amino acid), LC-FR3 (or a variant thereof in which one, two, or three amino acids in LC-FR3 are substituted with another amino acid), and LC-FR4 (or a variant thereof in which one, two, or three amino acids in LC-FR4 are substituted with another amino acid), wherein the LC-FR1, LC-FR2, LC-FR3, and LC-FR4 sequences in column B are selected from the same row of Table B. Contains the VL region.

[0093] In some embodiments, the antigen binding molecule comprises: LC-CDR1 (or 1 or 2 or 3 amino acids in LC-CDR1) shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column B of Table A LC-CDR1 (or a variant thereof in which one, two, or three amino acids in LC-CDR2 are substituted with another amino acid), LC-CDR2 (or a variant thereof in which one, two, or three amino acids in LC-CDR2 are substituted with another amino acid), and LC-CDR3 (or a variant thereof in which one, two, or three amino acids in LC-CDR3 are substituted with another amino acid), wherein the LC-CDR1, LC-CDR2, and LC-CDR3 sequences in column B are selected from the same row of Table A; and LC-FR1 (or a variant thereof in which one, two or three amino acids in LC-FR1 are substituted with other amino acids) or LC-FR2 shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column B of Table B (or a variant thereof in which one, two, or three amino acids in LC-FR2 are substituted with another amino acid), LC-FR3 (or a variant thereof in which one, two, or three amino acids in LC-FR3 are substituted with another amino acid), and LC-FR4 (or a variant thereof in which one, two, or three amino acids in LC-FR4 are substituted with another amino acid), wherein the LC-FR1, LC-FR2, LC-FR3, and LC-FR4 sequences in column B are selected from the same row of Table B. The VL region comprises:

[0094] In some embodiments, the antigen binding molecule comprises: LC-CDR1 (or LC-CDR2) as shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column B of Table A. - LC-CDR1 (or a variant thereof in which 1, 2 or 3 amino acids in LC-CDR2 are replaced by another amino acid), LC-CDR2 (or a variant thereof in which 1, 2 or 3 amino acids in LC-CDR2 are replaced by another amino acid) and LC-CDR3 (or a variant thereof in which 1, 2 or 3 amino acids in LC-CDR3 are replaced by another amino acid); and LC-FR1 (or LC-FR1 in which one or two or three amino acids are not present in another) shown in rows 1, 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, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50 or 51 of column B of Table B LC-FR2 (or a variant thereof in which one, two or three amino acids in LC-FR2 are substituted with another amino acid), LC-FR3 (or a variant thereof in which one, two or three amino acids in LC-FR3 are substituted with another amino acid), and LC-FR4 (or a variant thereof in which one, two or three amino acids in LC-FR4 are substituted with another amino acid). wherein the LC-CDR1, LC-CDR2, LC-CDR3 sequences of column B of Table A and the LC-FR1, LC-FR2, LC-FR3 and LC-FR4 sequences of column B of Table B are selected from rows having the same number, Contains the VL region.

[0095] In some embodiments, the antigen binding molecule is selected from the group consisting of: The VL region has at least 70%, preferably ≥80%, ≥85%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100% amino acid sequence identity to the amino acid sequence of a VL region sequence selected from 46, 47, 48, 49, 50 or 51.

[0096] In some embodiments, the antigen binding molecule comprises a VH region according to any one of the embodiments described herein and a VL region according to any one of the embodiments described herein.

[0097] In embodiments according to the present disclosure, one or more amino acids are substituted with another amino acid. Substitutions include replacing an amino acid residue with a non-identical "replacement" amino acid residue. The replacement amino acid residue of a substitution according to the present disclosure may be a naturally occurring amino acid residue (i.e., encoded by the genetic code) that is not identical to the amino acid residue at the relevant position in the corresponding unsubstituted amino acid sequence: alanine (Ala), arginine (Arg), asparagine (Asn), aspartic acid (Asp), cysteine ​​(Cys), glutamine (Gln), glutamic acid (Glu), glycine (Gly), histidine (His), isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), phenylalanine (Phe), proline (Pro), serine (Ser), threonine (Thr), tryptophan (Trp), tyrosine (Tyr), and valine (Val). In some embodiments, the replacement amino acid may be a non-naturally occurring amino acid residue—i.e., an amino acid residue other than those listed in the preceding sentence. Examples of non-naturally occurring amino acid residues include norleucine, ornithine, norvaline, homoserine, aib, and other amino acid residue analogs such as those described in Ellman et al., Meth. Enzym. 202 (1991) 301-336.

[0098] In some embodiments, substitutions may be biochemically conservative. In some embodiments, when a substituted amino acid is provided in one of rows 1 to 5 of the table below, the replacement amino acid of the substitution is another, non-identical amino acid provided in the same row:

[0099] [Table 2]

[0100] By way of example, in some embodiments, when the substitution is for a Met residue, the replacement amino acid may be selected from Ala, Val, Leu, lie, Trp, Tyr, Phe, and norleucine.

[0101] In some embodiments, the replacement amino acid in a substitution may have the same side chain polarity as the amino acid residue it replaces. In some embodiments, the replacement amino acid in a substitution may have the same side chain charge (at pH 7.4) as the amino acid residue it replaces:

[0102] [Table 3]

[0103] That is, in some embodiments, a non-polar amino acid is substituted with another, non-identical non-polar amino acid; in some embodiments, a polar amino acid is substituted with another, non-identical polar amino acid; in some embodiments, an acidic polar amino acid is substituted with another, non-identical acidic polar amino acid; in some embodiments, a basic polar amino acid is substituted with another, non-identical basic polar amino acid; in some embodiments, a neutral amino acid is substituted with another, non-identical neutral amino acid; in some embodiments, a positively charged amino acid is substituted with another, non-identical positively charged amino acid; in some embodiments, a negatively charged amino acid is substituted with another, non-identical negatively charged amino acid.

[0104] In some embodiments, the substitution(s) may be functionally conservative, i.e., in some embodiments, the substitution may not affect (or may not substantially affect) one or more functional properties (e.g., target binding) of an antigen-binding molecule comprising the substitution, compared to the corresponding unsubstituted molecule.

[0105] The VH and VL regions of the antigen-binding region of an antibody together constitute an Fv region. In some embodiments, an antigen-binding molecule according to the present disclosure comprises or consists of an Fv region that binds to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). In some embodiments, the VH and VL regions of the Fv are provided as a single polypeptide, i.e., a single-chain Fv (scFv), linked by a linker sequence.

[0106] The VL and light chain constant (CL) regions and the VH and heavy chain constant 1 (CH1) regions of the antigen-binding region of an antibody together constitute a Fab region. In some embodiments, an antigen-binding molecule comprises a Fab region comprising VH, CH1, VL, and CL (e.g., CK or Cλ). In some embodiments, the Fab region comprises a polypeptide comprising VH and CH1 (e.g., a VH-CH1 fusion polypeptide) and a polypeptide comprising VL and CL (e.g., a VL-CL fusion polypeptide). In some embodiments, the Fab region comprises a polypeptide comprising VH and CL (e.g., a VH-CL fusion polypeptide) and a polypeptide comprising VL and CH (e.g., a VL-CH1 fusion polypeptide); i.e., in some embodiments, the Fab region is a Cross-Fab region. In some embodiments, the VH, CH1, VL, and CL regions of a Fab or Cross-Fab are provided as a single polypeptide linked by a linker region, i.e., as a single-chain Fab (scFab) or single-chain Cross-Fab (scCrossFab).

[0107] In some embodiments, the antigen-binding molecules described herein comprise or consist of a whole antibody that binds to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). As used herein, "whole antibody" refers to an antibody having a structure substantially similar to that of an immunoglobulin (Ig). Various types of immunoglobulins and their structures are described, for example, in Schroeder and Cavacini J Allergy Clin Immunol. (2010) 125(202):S41-S52, which is incorporated herein by reference in its entirety.

[0108] G-type immunoglobulins (i.e., IgG) are glycoproteins of approximately 150 kDa that contain two heavy chains and two light chains. From the N- to C-terminus, the heavy chain contains a VH followed by a heavy chain constant region containing three constant domains (CH1, CH2, and CH3), and similarly, the light chain contains a VL followed by a CL. Depending on the heavy chain, immunoglobulins can be classified as IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM. The light chain can be kappa (κ) or lambda (λ).

[0109] As used herein, "CH1 domain" refers to the amino acid sequence corresponding to the CH1 domain of an immunoglobulin (Ig). The CH1 domain is the region of an Ig formed by positions 118 to 215 of the immunoglobulin constant domain according to the EU numbering system (described in Edelman et al., Proc Natl Acad Sci USA (1969) 63(1):78-85). "Hinge domain" refers to the amino acid sequence corresponding to the hinge domain of an Ig. The hinge domain is the region of an Ig formed by positions 216 to 230 of the immunoglobulin constant domain according to the EU numbering system. "CH2 domain" refers to the amino acid sequence corresponding to the CH2 domain of an Ig. The CH2 domain is the region of an Ig formed by positions 231 to 340 of the immunoglobulin constant domain according to the EU numbering system. "CH3 domain" refers to the amino acid sequence corresponding to the CH3 domain of an immunoglobulin (Ig). The CH3 domain is the region of an Ig formed by positions 341 to 447 of the immunoglobulin constant domain according to the EU numbering system. "CH2-CH3 region" refers to the amino acid sequence corresponding to the CH2 and CH3 domains of an immunoglobulin (Ig). The CH2-CH3 region is the region of an Ig formed by positions 231 to 447 of the immunoglobulin constant domain according to the EU numbering system.

[0110] In some embodiments, the antigen binding molecules described herein comprise or consist of an IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM that binds to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins).

[0111] In some embodiments, an antigen-binding molecule of the present disclosure comprises one or more regions (e.g., CH1, CH2, CH3, etc.) of an immunoglobulin heavy chain constant sequence. In some embodiments, the immunoglobulin heavy chain constant sequence is or is derived from the heavy chain constant sequence of IgG (e.g., IgG1, IgG2, IgG3, IgG4), IgA (e.g., IgA1, IgA2), IgD, IgE, or IgM, such as human IgG (e.g., hIgG1, hIgG2, hIgG3, hIgG4), hIgA (e.g., hIgA1, hIgA2), hIgD, hIgE, or hIgM. In some embodiments, the immunoglobulin heavy chain constant sequence is or is derived from the heavy chain constant sequence of a human IgG1 allotype (e.g., G1m1, G1m2, G1m3, or G1m17).

[0112] In some embodiments, the antigen-binding molecule comprises a CH1 region (e.g., comprises one or more polypeptides comprising a CH1 region). In some embodiments, the CH1 region comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 671 or 676.

[0113] In some embodiments, the antigen-binding molecule comprises a hinge region (e.g., comprises one or more polypeptides comprising a hinge region). In some embodiments, the hinge region comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 672.

[0114] In some embodiments, the antigen-binding molecule comprises a CH2 region (e.g., comprises one or more polypeptides comprising a CH2 region). In some embodiments, the CH2 region comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 673.

[0115] In some embodiments, the antigen-binding molecule comprises a CH3 region (e.g., comprises one or more polypeptides comprising a CH3 region). In some embodiments, the CH3 region comprises or consists of an amino acid sequence having at least 70% sequence identity (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) to the amino acid sequence of SEQ ID NO: 674 or 677.

[0116] In some embodiments, an antigen binding molecule of the present disclosure comprises an Fc region. As used herein, "Fc region" refers to the polypeptide complex formed by the interaction between two polypeptides, each of which contains the CH2-CH3 region of an immunoglobulin (Ig) heavy chain constant sequence.

[0117] In some embodiments, a CH2 region, a CH3 region, and / or a CH2-CH3 region according to the present disclosure corresponds to the CH2 region / CH3 region / CH2-CH3 region of an IgG (e.g., IgG1, IgG2, IgG3, IgG4), an IgA (e.g., IgA1, IgA2), an IgD, an IgE, or an IgM. In some embodiments, a CH2 region, a CH3 region, and / or a CH2-CH3 region corresponds to the CH2 region / CH3 region / CH2-CH3 region of a human IgG (e.g., hIgG1, hIgG2, hIgG3, hIgG4), an hIgA (e.g., hIgA1, hIgA2), an hIgD, an hIgE, or an hIgM. In some embodiments, the CH2 region, CH3 region and / or CH2-CH3 region correspond to the CH2 region / CH3 region / CH2-CH3 region of a human IgG1 allotype (eg, G1m1, G1m2, G1m3 or G1m17).

[0118] The Fc region provides interaction with Fc receptors and other molecules of the immune system to produce functional effects. Fc-mediated effector function is outlined, for example, in Jefferis et al., Immunol Rev 1998 163:59-76 (incorporated herein by reference in its entirety), and is achieved through the Fc-mediated recruitment and activation of immune cells (e.g., macrophages, dendritic cells, neutrophils, basophils, eosinophils, platelets, mast cells, NK cells, and T cells) through the interaction between the Fc region and Fc receptors expressed by immune cells, the recruitment of components of the complement pathway through the binding of the Fc region to the complement protein C1q, and the resulting activation of the complement cascade. Fc-mediated functions include Fc receptor binding, antibody-dependent cellular cytotoxicity (ADCC), antibody-dependent cell-mediated phagocytosis (ADCP), complement-dependent cytotoxicity (CDC), membrane attack complex formation (MAC), cell degranulation, cytokine and / or chemokine production, and antigen processing and presentation.

[0119] Modifications to antibody Fc regions that affect Fc-mediated function are known in the art, such as those described in Wang et al., Protein Cell (2018) 9(1):63-73 (incorporated herein by reference in its entirety). Exemplary Fc region modifications known to affect antibody effector function are summarized in Table 1 of Wang et al., Protein Cell (2018) 9(1):63-73. In some embodiments, antigen-binding molecules of the present disclosure comprise an Fc region comprising a modification that increases or decreases an Fc-mediated function compared to an antigen-binding molecule comprising a corresponding unmodified Fc region. When the Fc region comprises a modification, the modification may be present in one or both of the polypeptide chains that together form the Fc region.

[0120] In some embodiments, the antigen-binding molecule comprises a CH2-CH3 region (e.g., comprises one or more polypeptides comprising a CH2-CH3 region). In some embodiments, the CH2-CH3 region comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 678 or 679.

[0121] In some embodiments, the antigen-binding molecule comprises a CH1-hinge-CH2-CH3 region (e.g., comprises one or more polypeptides comprising a CH1-hinge-CH2-CH3 region). In some embodiments, the CH1-hinge-CH2-CH3 region comprises or consists of an amino acid sequence having at least 70% (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) sequence identity to the amino acid sequence of SEQ ID NO: 670.

[0122] In some embodiments, the antigen-binding molecule of the present disclosure comprises one or more regions of an immunoglobulin light chain constant sequence. In some embodiments, the immunoglobulin light chain constant sequence is a human immunoglobulin kappa constant (IGKC; Cκ). In some embodiments, the immunoglobulin light chain constant sequence is a human immunoglobulin lambda constant (IGLC; Cλ), such as IGLC1, IGLC2, IGLC3, IGLC6, or IGLC7.

[0123] In some embodiments, the antigen-binding molecule comprises a CL region (e.g., comprises one or more polypeptides comprising a CL region). In some embodiments, the CL region comprises or consists of an amino acid sequence having at least 70% sequence identity (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99%, or 100%) to the amino acid sequence of SEQ ID NO: 680, 681, 682, 683, 684, or 685.

[0124] In some embodiments, the antigen-binding molecule is or comprises a monoclonal antibody or an antigen-binding fragment thereof. In some embodiments, the antigen-binding molecule is or comprises a fully human antibody / antibody fragment. A fully human antibody / antibody fragment may be encoded by a human nucleic acid sequence(s). A fully human antibody / antibody fragment may not have non-human amino acid sequences.

[0125] Aspects of the present disclosure relate to multispecific antigen-binding molecules. By "multispecific," it is meant that an antigen-binding molecule exhibits specific binding to more than one target. In some embodiments, the antigen-binding molecule is a bispecific antigen-binding molecule. In some embodiments, the antigen-binding molecule comprises at least two different antigen-binding domains (i.e., at least two antigen-binding domains, for example, comprising non-identical VH and VL).

[0126] In some embodiments, the antigen-binding molecule binds to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) and another target other than the sarbecovirus spike protein, and is therefore at least bispecific. The term "bispecific" means that the antigen-binding molecule can specifically bind to at least two distinct antigenic determinants.

[0127] It will be understood that antigen-binding molecules (e.g., multispecific antigen-binding molecules) according to the present disclosure may include antigen-binding molecules capable of binding to a target for which the antigen-binding molecule is specific. For example, antigen-binding molecules that bind to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) and a target other than the sarbecovirus spike protein may include (i) antigen-binding molecules that bind to the SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins, and (ii) antigen-binding molecules that bind to an antigen other than the sarbecovirus spike protein.

[0128] It will also be understood that an antigen-binding molecule (e.g., a multispecific antigen-binding molecule) according to the present disclosure may comprise an antigen-binding polypeptide or antigen-binding polypeptide complex capable of binding to a target for which the antigen-binding molecule is specific.

[0129] In some embodiments, an antigen-binding molecule that is a component of a larger antigen-binding molecule (e.g., a multispecific antigen-binding molecule) may be referred to, for example, as an "antigen-binding domain" or "antigen-binding region" of the larger antigen-binding molecule.

[0130] In some embodiments, the antigen-binding molecule is an immune cell engager. Immune cell engagers are reviewed, for example, in Goebeler and Bargou, Nat. Rev. Clin. Oncol. (2020) 17:418-434 and Ellerman, Methods (2019) 154:102-117, both of which are incorporated herein by reference in their entireties. Immune cell engager molecules include an antigen-binding region for a target antigen of interest and an antigen-binding region for recruiting / engaging immune cells of interest. Immune cell engagers recruit / engage immune cells through antigen-binding regions specific for immune cell surface molecules.

[0131] The most well-studied immune cell engagers are bispecific T cell engagers (BiTEs), which contain a target antigen-binding domain and a CD3 polypeptide (typically, CD3ε)-binding domain through which BiTEs recruit T cells. Binding of a BiTE to its target antigen and to a CD3 polypeptide expressed by a T cell results in T cell activation and ultimately directs T cell effector activity against cells expressing the target antigen. Other types of immune cell engagers are known in the art, including natural killer cell engagers, e.g., bispecific killer engagers (BiKEs), which activate and recruit NK cells.

[0132] In some embodiments, the immune cells engaged by the immune cell engager are T cells or NK cells, hi some embodiments, the immune cell engager is a T cell engager.

[0133] Multispecific antigen-binding molecules according to the present disclosure may be provided in any suitable format, such as those described in Brinkmann and Kontermann, MAbs (2017) 9(2):182-212, which is incorporated herein by reference in its entirety. Suitable formats include those described in Figure 2 of Brinkmann and Kontermann, MAbs (2017) 9(2):182-212: antibody conjugates, such as IgG2, F(ab')2, or CovX-Body; IgG or IgG-like molecules, such as IgG, chimeric IgG, κλ-body common HC; CH1 / CL fusion proteins, such as scFv2-CH1 / CL, VHH2-CH1 / CL; "variable domain only" bispecific antigen binding molecules, such as tandem scFv (taFv), triplebody, diabody (Db), dsDb, Db(kih), DART, scDB, dsFv-dsFv, tandAbs, triple heads, tandem dAb / VHH, tertravalent dAb.VHH); non-Ig fusion proteins, e.g., scFv2-albumin, scDb-albumin, taFv-albumin, taFv-toxin, miniantibodies, DNL-Fab2, DNL-Fab2-scFv, DNL-Fab2-IgG-cytokine2, ImmTAC (TCR-scFv); modified Fc and CH3 fusion proteins, e.g., scFv-Fc(kih), scFv-Fc(CH3 charge pair), scFv-Fc(EW-RVT), scFv-fc(HA-TF), scFv-Fc(SEEDbody), taFv-Fc(kih), sc Fv-Fc(kih)-Fv, Fab-Fc(kih)-scFv, Fab-scFv-Fc(kih), Fab-scFv-Fc(BEAT), Fab-scFv-Fc(SEEDbody), DART-Fc, scFv-CH3(kih), TriFabs; Fc fusions, e.g., Di-Diabody, scDb-Fc, taFv-Fc, scFv-Fc-scFv, HCAb-VHH, Fab-scFv-Fc, scFv4-Ig, scFv2-Fcab; CH3 fusions, e.g., Dia-Diabody, scDb-CH3; IgE / IgM CH2 fusions, e.g., scFv-EHD2-scFv, scFvMHD2-scFv; Fab fusion proteins, e.g., Fab-scFv (bibody), Fab-scFv2 (tribody), Fab-Fv, Fab-dsFv, Fab-VHH, orthogonal Fab-Fab; non-Ig fusion proteins, e.g., DNL-Fab3, DNL-Fab2-scFv, DNL-Fab2-IgG-cytokine2; asymmetric IgG or IgG-like molecules, e.g., IgG(kih), IgG(kih) common LC, ZW1 IgG common LC, Biclonics common LC, CrossMab, CrossMab(kih), scFab-IgG(kih), Fab-scFab-IgG(kih), orthogonal Fab IgG(kih), DuetMab, CH3 charged pair + CH1 / CL charged pair, hinge / CH3 charged pair, SEED-body, Duobody, four-in-one-CrossMab(kih), LUZ-Y common LC; LUZ-Y scFab-IgG, FcFc. *; adducted and Fc-modified IgG, e.g., IgG(kih)-Fv, IgG HA-TF-Fv, IgG(kih)scFab, scFab-Fc(kih)-scFv2, scFab-Fc(kih)-scFv, half DVD-Ig, DVI-Ig (four-in-one), CrossMab-Fab; modified Fc and CH3 fusion proteins, e.g., Fab-Fc(kih)-scFv, Fab-scFv-Fc(kih), Fab-scFv-Fc(BEAT), Fab-scFv-Fc-SEEDbody, TriFab; adducted IgG-HC fusions, e.g., IgG-HC, scFv, IgG-dAb, IgG-taFV, IgG-CrossFab, IgG-orthogonal Fab, IgG-(CαCβ)Fab, scFv-HC-IgG, tandem Fab-IgG (orthogonal Fab), Fab-IgG(CαCβ) Fab), Fab-IgG(CR3), Fab-hinge-IgG(CR3); additional IgG-LC fusions, e.g., IgG-scFv(LC), scFv(LC)-IgG, dAb-IgG; additional IgG-HC and LC fusions, e.g., DVD-Ig, TVD-Ig, CODV-Ig, scFv4-IgG, Zybody; Fc fusions, e.g., Fab-scFv-Fc, scFv4-Ig; F(ab')2 fusions, e.g., F(ab')2-scFv2; CH1 / CL fusion proteins, e.g., scFv2-CH1-hinge / CL; modified IgGs, e.g., DAF (two-in-one-IgG), DutaMab, Mab 2 and non-Ig fusions, such as DNL-Fab4-IgG. Multispecific antigen-binding molecules can be readily designed and produced by those skilled in the art.

[0134] The present disclosure also provides chimeric antigen receptors (CARs). CARs are recombinant receptors that provide both antigen binding and T cell activation functions. CAR structure and operation are outlined, for example, in Dotti et al., Immunol Rev (2014) 257 (1), the entire contents of which are incorporated herein by reference. CARs comprise an antigen binding region linked to a cell membrane anchor region and a signal transduction region. An optional hinge region can provide separation between the antigen binding region and the cell membrane anchor region and can act as a flexible linker.

[0135] The antigen-binding domain of a CAR according to the present disclosure comprises or consists of an antigen-binding molecule that binds to a sarbecovirus spike protein described herein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). Thus, a CAR according to the present disclosure comprises an antigen-binding molecule described herein.

[0136] It will be understood that the antigen-binding molecule according to the present disclosure forms or is included in the antigen-binding domain of the CAR. Thus, in some embodiments, the antigen-binding molecule of the present disclosure is included in the CAR.

[0137] It will also be understood that an antigen-binding molecule according to the present disclosure can be a CAR. A CAR having an antigen-binding domain comprising or consisting of an antigen-binding molecule of the present disclosure (e.g., a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins)-binding Fv) is an antigen-binding molecule. The antigen-binding domain of a CAR of the present disclosure may be provided in any suitable format, such as an scFv, scFab, etc.

[0138] The cell membrane anchor region is provided between the antigen binding region and the signal transduction region of the CAR, and provides anchoring of the CAR to the cell membrane of a cell expressing the CAR, with the antigen binding region in the extracellular space and the signal transduction region intracellularly. In some embodiments, the CAR comprises a cell membrane anchor region that comprises or consists of an amino acid sequence that comprises, consists of, or is derived from the transmembrane region amino acid sequence of one of CD3-zeta, CD4, CD8, or CD28. As used herein, a region that is "derived" from a reference amino acid sequence comprises an amino acid sequence that has at least 60%, e.g., at least 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the reference sequence.

[0139] The signaling region of a CAR enables T cell activation. The CAR signaling region may contain the amino acid sequence of the intracellular domain of CD3-zeta, which provides an immunoreceptor tyrosine-based activation motif (ITAM) for phosphorylation and activation of CAR-expressing T cells. Signaling regions containing sequences from other ITAM-containing proteins, such as FcγRI, are also used in CARs (Haynes et al., 2001 J Immunol 166(1):182-187). The signaling region of a CAR may also contain a costimulatory sequence derived from the signaling region of a costimulatory molecule to promote activation of CAR-expressing T cells upon binding to a target protein. Suitable costimulatory molecules include CD28, OX40, 4-1BB, ICOS, and CD27. In some cases, CARs are engineered to provide costimulation of different intracellular signaling pathways. For example, signaling associated with CD28 costimulation preferentially activates the phosphatidylinositol 3-kinase (PI3K) pathway, while 4-1BB-mediated signaling uses TNF receptor-associated factor (TRAF) adaptor proteins. Thus, the signaling region of a CAR sometimes contains costimulatory sequences derived from the signaling region of more than one costimulatory molecule. In some embodiments, a CAR of the present disclosure comprises one or more costimulatory sequences that comprise, consist of, or are derived from the amino acid sequence of one or more intracellular domains of CD28, OX40, 4-1BB, ICOS, and CD27.

[0140] The optional hinge region can provide separation between the antigen-binding domain and the transmembrane domain and can act as a flexible linker. The hinge region can be derived from IgG1 or IgG4. In some embodiments, the CAR of the present disclosure comprises a hinge region that comprises, consists of, or is derived from the amino acid sequence of the hinge region of IgG1 or IgG4.

[0141] Similarly, cells comprising CARs according to the present disclosure are provided. CARs according to the present disclosure can be used to generate CAR-expressing immune cells, such as CAR-T or CAR-NK cells. The manipulation of CARs into immune cells can be carried out in vitro during culture.

[0142] Functional properties of the antigen-binding molecules of the present disclosure The antigen-binding molecules described herein may be characterized by certain functional properties. In some embodiments, the antigen-binding molecules described herein have the following properties: binds to a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins); inhibiting the interaction between a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) and ACE2; and / or Inhibiting infection of ACE2-expressing cells by sarbecoviruses (e.g., SARS-CoV; e.g., SARS-CoV-2 and / or one or more SARS-CoV-2 variants) may have one or more of:

[0143] It will be understood that a given antigen-binding molecule may exhibit more than one of the properties listed in the preceding paragraph. A given antigen-binding molecule can be evaluated for the properties listed in the preceding paragraph using a suitable assay. For example, the assay can be, for example, an in vitro assay, and optionally a cell-based or cell-free assay. In some embodiments, the assay can be, for example, an in vivo assay, i.e., performed in a non-human animal. In some embodiments, the assay can be an ex vivo assay, i.e., performed using cells / tissues / organs obtained from a subject.

[0144] When the assays are cell-based assays, they may involve treating cells with a given antigen-binding molecule to determine whether the antigen-binding molecule exhibits one or more of the listed properties. The assays may use molecular species labeled with a detectable entity to facilitate detection of the molecular species. The assays may involve evaluating the listed properties after treating the cells separately with various amounts / concentrations (e.g., serial dilutions) of the given antigen-binding molecule. It will be understood that the cells preferably express the target antigen for the antigen-binding molecule (i.e., a sarbecovirus spike protein (e.g., a SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein)).

[0145] Analysis of the results of such assays may include determining the concentration at which 50% of the maximal level of relevant activity is achieved. The concentration of a given drug at which 50% of the maximal level of relevant activity is achieved may be referred to as the "half-maximal effective concentration" of the drug in relation to the relevant activity, or the "EC 50 It is sometimes called ".

[0146] Depending on the characteristics, EC 50is the "half-maximal inhibitory concentration" or "IC 50 " is the concentration of an agent at which 50% of the maximal level of inhibition of a given property is observed.

[0147] The antigen binding molecules described herein bind to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins).

[0148] The ability of a given antigen-binding molecule to specifically bind to a given peptide / polypeptide can be determined by analysis using methods known in the art, such as ELISA, surface plasmon resonance (SPR; see, e.g., Hearty et al., Methods Mol Biol (2012) 907:411-442), biolayer interferometry (BLI; see, e.g., Lad et al., (2015) J Biomol Screen 20(4):498-507), flow cytometry, or radiolabeled antigen binding assay (RIA) enzyme-linked immunosorbent assay. Through these analyses, binding to a given molecule can be measured and quantified. In some embodiments, binding can be a response detected in a given assay.

[0149] In some embodiments, the antigen-binding molecule according to the present disclosure binds to the SARS-CoV-2 spike protein. In some embodiments, the antigen-binding molecule according to the present disclosure binds to a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1.

[0150] In some embodiments, an antigen-binding molecule according to the present disclosure binds to a SARS-CoV-2 variant spike protein described herein. In some embodiments, an antigen-binding molecule according to the present disclosure binds to a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO: 1, where the amino acid sequence is not identical to SEQ ID NO: 1. In some embodiments, an antigen-binding molecule according to the present disclosure binds to a polypeptide comprising or consisting of SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, or 27.

[0151] In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of SEQ ID NO: 18. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of SEQ ID NO: 19. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of SEQ ID NO: 20. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of SEQ ID NO: 21. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of SEQ ID NO: 26. In some embodiments, the antigen-binding molecule binds to a polypeptide comprising or consisting of SEQ ID NO: 27.

[0152] In some embodiments, an antigen-binding molecule according to the present disclosure binds to a polypeptide comprising an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO: 7, wherein the amino acid sequence is not identical to SEQ ID NO: 7. In some embodiments, an antigen-binding molecule according to the present disclosure binds to a polypeptide comprising or consisting of SEQ ID NO: 686, 687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 699, 700, 701, or 702.

[0153] In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 693. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 694. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 695. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 696. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 697. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 698.

[0154] In some embodiments, an antigen-binding molecule according to the present disclosure binds to a polypeptide comprising an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO: 8, wherein the amino acid sequence is not identical to SEQ ID NO: 8. In some embodiments, an antigen-binding molecule according to the present disclosure binds to a polypeptide comprising or consisting of SEQ ID NO: 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, or 719.

[0155] In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 710. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 711. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 712. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 713. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 714. In some embodiments, the antigen binding molecule binds to a polypeptide comprising SEQ ID NO: 715.

[0156] In some embodiments, an antigen-binding molecule according to the present disclosure can (independently) bind to two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more) proteins selected from SARS-CoV-2 spike protein and SARS-CoV-2 variant spike protein. That is, in some embodiments, an antigen-binding molecule that binds to a given (first) protein selected from SARS-CoV-2 spike protein and SARS-CoV-2 variant spike protein also binds to one or more additional (second, third, etc.) proteins selected from SARS-CoV-2 spike protein and SARS-CoV-2 variant spike protein, wherein the one or more additional proteins have an amino acid sequence that differs from the amino acid sequence of the first protein. Such antigen-binding molecules may be described as being "cross-reactive" with respect to the first protein and the additional protein, or may be said to exhibit "cross-reactivity" or "cross-reactive binding" or "bind cross-reactively" to the first protein and the additional protein.

[0157] In some embodiments, an antigen-binding molecule according to the present disclosure cross-reactively binds to two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or all 18) polypeptides selected from the following: a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 11; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 13; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 14; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 15; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 16; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 17; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 18; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 19; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 20; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 21; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 22; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 23; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 24; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 25; a polypeptide consisting of the amino acid sequence of SEQ ID NO: 26; and a polypeptide consisting of the amino acid sequence of SEQ ID NO: 27.

[0158] In some embodiments, the antigen-binding molecule according to the present disclosure is a polypeptide comprising the amino acid sequence of SEQ ID NO: 7, a polypeptide comprising the amino acid sequence of SEQ ID NO: 686, a polypeptide comprising the amino acid sequence of SEQ ID NO: 687, a polypeptide comprising the amino acid sequence of SEQ ID NO: 688, a polypeptide comprising the amino acid sequence of SEQ ID NO: 689, a polypeptide comprising the amino acid sequence of SEQ ID NO: 690, a polypeptide comprising the amino acid sequence of SEQ ID NO: 691, a polypeptide comprising the amino acid sequence of SEQ ID NO: 692, a polypeptide comprising the amino acid sequence of SEQ ID NO: 693, a polypeptide comprising the amino acid sequence of SEQ ID NO: 694, or a polypeptide comprising the amino acid sequence of SEQ ID NO: 695. a polypeptide comprising the amino acid sequence of SEQ ID NO: 696, a polypeptide comprising the amino acid sequence of SEQ ID NO: 697, a polypeptide comprising the amino acid sequence of SEQ ID NO: 698, a polypeptide comprising the amino acid sequence of SEQ ID NO: 699, a polypeptide comprising the amino acid sequence of SEQ ID NO: 700, a polypeptide comprising the amino acid sequence of SEQ ID NO: 701, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 702.

[0159] In some embodiments, the antigen-binding molecule according to the present disclosure is a polypeptide comprising the amino acid sequence of SEQ ID NO: 8, a polypeptide comprising the amino acid sequence of SEQ ID NO: 703, a polypeptide comprising the amino acid sequence of SEQ ID NO: 704, a polypeptide comprising the amino acid sequence of SEQ ID NO: 705, a polypeptide comprising the amino acid sequence of SEQ ID NO: 706, a polypeptide comprising the amino acid sequence of SEQ ID NO: 707, a polypeptide comprising the amino acid sequence of SEQ ID NO: 708, a polypeptide comprising the amino acid sequence of SEQ ID NO: 709, a polypeptide comprising the amino acid sequence of SEQ ID NO: 710, a polypeptide comprising the amino acid sequence of SEQ ID NO: 711, or a polypeptide comprising the amino acid sequence of SEQ ID NO: 712. a polypeptide comprising the amino acid sequence of SEQ ID NO: 713, a polypeptide comprising the amino acid sequence of SEQ ID NO: 714, a polypeptide comprising the amino acid sequence of SEQ ID NO: 715, a polypeptide comprising the amino acid sequence of SEQ ID NO: 716, a polypeptide comprising the amino acid sequence of SEQ ID NO: 717, a polypeptide comprising the amino acid sequence of SEQ ID NO: 718, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 719.

[0160] In a preferred embodiment, the antigen-binding molecule according to the present disclosure binds (i.e., cross-reactively) to a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 11, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 12, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 13, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 14, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 15, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 16, and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 17. In a preferred embodiment, the antigen-binding molecule according to the present disclosure binds (i.e., cross-reactively) to a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 11, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 12, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 13, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 14, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 15, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 16, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 17, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 18, and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 19.In a preferred embodiment, the antigen-binding molecule according to the present disclosure binds (i.e., cross-reactively) to a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 11, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 12, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 13, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 14, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 15, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 16, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 17, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 18, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 19, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 20, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 21, a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 26, and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 27.

[0161] In a preferred embodiment, an antigen-binding molecule according to the present disclosure binds (i.e., by cross-reactivity) to a polypeptide comprising the amino acid sequence of SEQ ID NO: 7, a polypeptide comprising the amino acid sequence of SEQ ID NO: 686, a polypeptide comprising the amino acid sequence of SEQ ID NO: 687, a polypeptide comprising the amino acid sequence of SEQ ID NO: 688, a polypeptide comprising the amino acid sequence of SEQ ID NO: 689, a polypeptide comprising the amino acid sequence of SEQ ID NO: 690, a polypeptide comprising the amino acid sequence of SEQ ID NO: 691, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 692. In a preferred embodiment, an antigen-binding molecule according to the present disclosure binds (i.e., by cross-reactivity) to a polypeptide comprising the amino acid sequence of SEQ ID NO: 7, a polypeptide comprising the amino acid sequence of SEQ ID NO: 686, a polypeptide comprising the amino acid sequence of SEQ ID NO: 687, a polypeptide comprising the amino acid sequence of SEQ ID NO: 688, a polypeptide comprising the amino acid sequence of SEQ ID NO: 689, a polypeptide comprising the amino acid sequence of SEQ ID NO: 690, a polypeptide comprising the amino acid sequence of SEQ ID NO: 691, a polypeptide comprising the amino acid sequence of SEQ ID NO: 692, a polypeptide comprising the amino acid sequence of SEQ ID NO: 693, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 694. In a preferred embodiment, the antigen-binding molecule according to the present disclosure binds (i.e., cross-reactively) to a polypeptide comprising the amino acid sequence of SEQ ID NO: 7, a polypeptide comprising the amino acid sequence of SEQ ID NO: 686, a polypeptide comprising the amino acid sequence of SEQ ID NO: 687, a polypeptide comprising the amino acid sequence of SEQ ID NO: 688, a polypeptide comprising the amino acid sequence of SEQ ID NO: 689, a polypeptide comprising the amino acid sequence of SEQ ID NO: 690, a polypeptide comprising the amino acid sequence of SEQ ID NO: 691, a polypeptide comprising the amino acid sequence of SEQ ID NO: 692, a polypeptide comprising the amino acid sequence of SEQ ID NO: 693, a polypeptide comprising the amino acid sequence of SEQ ID NO: 694, a polypeptide comprising the amino acid sequence of SEQ ID NO: 695, a polypeptide comprising the amino acid sequence of SEQ ID NO: 696, a polypeptide comprising the amino acid sequence of SEQ ID NO: 697, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 698.

[0162] In a preferred embodiment, an antigen-binding molecule according to the present disclosure binds (i.e., by cross-reactivity) to a polypeptide comprising the amino acid sequence of SEQ ID NO: 8, a polypeptide comprising the amino acid sequence of SEQ ID NO: 703, a polypeptide comprising the amino acid sequence of SEQ ID NO: 704, a polypeptide comprising the amino acid sequence of SEQ ID NO: 705, a polypeptide comprising the amino acid sequence of SEQ ID NO: 706, a polypeptide comprising the amino acid sequence of SEQ ID NO: 707, a polypeptide comprising the amino acid sequence of SEQ ID NO: 708, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 709. In a preferred embodiment, an antigen-binding molecule according to the present disclosure binds (i.e., by cross-reactivity) to a polypeptide comprising the amino acid sequence of SEQ ID NO: 8, a polypeptide comprising the amino acid sequence of SEQ ID NO: 703, a polypeptide comprising the amino acid sequence of SEQ ID NO: 704, a polypeptide comprising the amino acid sequence of SEQ ID NO: 705, a polypeptide comprising the amino acid sequence of SEQ ID NO: 706, a polypeptide comprising the amino acid sequence of SEQ ID NO: 707, a polypeptide comprising the amino acid sequence of SEQ ID NO: 708, a polypeptide comprising the amino acid sequence of SEQ ID NO: 709, a polypeptide comprising the amino acid sequence of SEQ ID NO: 710, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 711. In a preferred embodiment, the antigen-binding molecule according to the present disclosure binds (i.e., cross-reactively) to a polypeptide comprising the amino acid sequence of SEQ ID NO: 8, a polypeptide comprising the amino acid sequence of SEQ ID NO: 703, a polypeptide comprising the amino acid sequence of SEQ ID NO: 704, a polypeptide comprising the amino acid sequence of SEQ ID NO: 705, a polypeptide comprising the amino acid sequence of SEQ ID NO: 706, a polypeptide comprising the amino acid sequence of SEQ ID NO: 707, a polypeptide comprising the amino acid sequence of SEQ ID NO: 708, a polypeptide comprising the amino acid sequence of SEQ ID NO: 709, a polypeptide comprising the amino acid sequence of SEQ ID NO: 710, a polypeptide comprising the amino acid sequence of SEQ ID NO: 711, a polypeptide comprising the amino acid sequence of SEQ ID NO: 712, a polypeptide comprising the amino acid sequence of SEQ ID NO: 713, a polypeptide comprising the amino acid sequence of SEQ ID NO: 714, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 715.

[0163] The antigen-binding molecules and antigen-binding domains described herein preferably exhibit specific binding to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). As used herein, "specific binding" refers to binding that is selective for the antigen and can be distinguished from nonspecific binding to non-target antigens. An antigen-binding molecule / domain that specifically binds to a target molecule preferably binds to the target with greater affinity and / or longer duration than it binds to other non-target molecules.

[0164] In some embodiments, the extent of binding of the antigen-binding molecule to the non-target molecule is less than about 10% of the binding of the antibody to the target molecule, as measured, for example, by ELISA, SPR, BLI, or by RIA. Alternatively, binding specificity can be measured by the dissociation constant (K D ) is at least 0.1 orders of magnitude larger (i.e., 0.1x10 n , where n is an integer representing the number of digits) dissociation constant (K D ) can be reflected in terms of the binding affinity with which the antigen-binding molecule binds, which can optionally be at least one of 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1.5, or 2.0.

[0165] In some embodiments, the antigen binding molecules described herein have affinities in the micromolar range, i.e., K D =9.9x10 -4 From 1x10 -6 In some embodiments, the antigen-binding molecules described herein bind to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) with a submicromolar affinity, i.e., a K D <1x10 -6In some embodiments, the antigen-binding molecules described herein bind to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) with an affinity in the nanomolar range, i.e., a K D =9.9x10 -7 From 1x10 -9 In some embodiments, the antigen-binding molecules described herein bind to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) with sub-nanomolar affinity, i.e., K D <1x10 -9 In some embodiments, the antigen-binding molecules described herein bind to a given SARS-CoV-2 spike protein with an affinity in the picomolar range, i.e., K D =9.9x10 -10 From 1x10 -12 In some embodiments, the antigen-binding molecules described herein bind to a sarbecovirus spike protein (e.g., a SARS-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) with a sub-picomolar affinity, i.e., a K D <1x10 -12 Binds to a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) at M.

[0166] The antigen-binding molecules of the present disclosure can bind to a specific region of interest of a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). The antigen-binding molecules of the present disclosure can bind to a linear epitope of a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) consisting of a contiguous sequence of amino acids (i.e., primary amino acid sequence). In some embodiments, the antigen-binding molecules can bind to a conformational epitope of a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) consisting of a non-contiguous sequence of amino acids in the amino acid sequence.

[0167] The region of a given target molecule to which an antigen-binding molecule binds can be determined using a variety of methods well known in the art, including X-ray co-crystallography analysis of antibody-antigen complexes, peptide scanning, mutagenesis mapping, hydrogen-deuterium exchange analysis by mass spectrometry, phage display, competitive ELISA, and proteolysis-based "protection" methods. Such methods are described, for example, in Gershoni et al., BioDrugs, 2007, 21(3):145-156, the entire contents of which are incorporated herein by reference.

[0168] In some embodiments, the antigen binding molecule can bind to a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) at a region that is the same as or overlaps with a region bound by an antibody, including the VH and VL regions of the antibodies shown in Table C.

[0169] Whether a test antigen-binding molecule binds to the same or overlapping region of a given target as a reference antigen-binding molecule can be evaluated, for example, by analyzing (i) the interaction between the test antigen-binding molecule and the target in the absence of the reference antigen-binding molecule, and (ii) the interaction between the test antigen-binding molecule in the presence of the reference antigen-binding molecule or after incubation of the target with the reference antigen-binding molecule. Determining a reduced level of interaction between the test antigen-binding molecule and the target after analysis by (ii) compared to (i) can support the inference that the test and reference antigen-binding molecules bind to the same or overlapping region of the target. Suitable assays for such analysis include, for example, competitive ELISA assays and epitope binning assays.

[0170] In some embodiments, the antigen binding molecule binds to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) in a region bound by an interaction partner for the protein, e.g., ACE2. In some embodiments, the antigen binding molecule reduces / inhibits the interaction between a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) and an interaction partner for the SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein(s) (e.g., ACE2). In some embodiments, the antigen binding molecule is a competitive inhibitor of the binding of an interaction partner for a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins), e.g., ACE2, to the sarbecovirus spike protein(s). In some embodiments, the antigen binding molecule binds to a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) in a region bound by a polypeptide comprising or consisting of the sequence set forth in SEQ ID NO:30.

[0171] Antigen-binding molecules that inhibit the interaction between ACE2 and a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) may be described as inhibitors / antagonists of such interaction, and may also be referred to as neutralizing antigen-binding molecules to a sarbecovirus (e.g., a SARSr-CoV, e.g., a SARS-CoV-2 and / or one or more SARS-CoV-2 variants).

[0172] In some embodiments, an antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and the SARS-CoV-2 spike protein. In some embodiments, an antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1.

[0173] In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a SARS-CoV-2 variant spike protein. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO:1, where the amino acid sequence is not identical to SEQ ID NO:1. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26 or 27.

[0174] In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 18. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 19. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 20. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 21. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 26. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 27.

[0175] In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO: 7, wherein the amino acid sequence is not identical to SEQ ID NO: 7. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 686, 687, 688, 689, 690, 691, 692, 693, 694, 695, 696, 697, 698, 699, 700, 701, or 702.

[0176] In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 693. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 694. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 695. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 696. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 697. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 698.

[0177] In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, or ≥99%) amino acid sequence identity to SEQ ID NO: 8, wherein the amino acid sequence is not identical to SEQ ID NO: 8. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising or consisting of SEQ ID NO: 703, 704, 705, 706, 707, 708, 709, 710, 711, 712, 713, 714, 715, 716, 717, 718, or 719.

[0178] In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 710. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 711. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 712. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 713. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 714. In some embodiments, the antigen binding molecule inhibits the interaction between ACE2 and a polypeptide comprising SEQ ID NO: 715.

[0179] In some embodiments, an antigen binding molecule according to the present disclosure can inhibit the interaction between ACE2 and two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, or more) proteins (independently) selected from SARS-CoV-2 spike protein and SARS-CoV-2 variant spike protein. That is, in some embodiments, an antigen binding molecule that inhibits the interaction between ACE2 and a given (first) protein selected from SARS-CoV-2 spike protein and SARS-CoV-2 variant spike protein also inhibits the interaction between ACE2 and one or more additional (second, third, etc.) proteins selected from SARS-CoV-2 spike protein and SARS-CoV-2 variant spike protein, where the one or more additional proteins have an amino acid sequence that differs from the amino acid sequence of the first protein.

[0180] In some embodiments, an antigen-binding molecule according to the present disclosure inhibits the interaction between ACE2 and two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or all 18) polypeptides selected from: a polypeptide consisting of the amino acid sequence of SEQ ID NO: 1, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 11, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 12, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 13, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 14, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 15, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 16, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 17, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 18, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 19, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 20, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 21, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 22, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 23, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 24, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 25, a polypeptide consisting of the amino acid sequence of SEQ ID NO: 26, and a polypeptide consisting of the amino acid sequence of SEQ ID NO: 27.

[0181] In some embodiments, the antigen binding molecule according to the present disclosure is capable of binding to ACE2: a polypeptide comprising the amino acid sequence of SEQ ID NO: 7, a polypeptide comprising the amino acid sequence of SEQ ID NO: 686, a polypeptide comprising the amino acid sequence of SEQ ID NO: 687, a polypeptide comprising the amino acid sequence of SEQ ID NO: 688, a polypeptide comprising the amino acid sequence of SEQ ID NO: 689, a polypeptide comprising the amino acid sequence of SEQ ID NO: 690, a polypeptide comprising the amino acid sequence of SEQ ID NO: 691, a polypeptide comprising the amino acid sequence of SEQ ID NO: 692, a polypeptide comprising the amino acid sequence of SEQ ID NO: 693, a polypeptide comprising the amino acid sequence of SEQ ID NO: 694, a polypeptide comprising the amino acid sequence of SEQ ID NO: 695 a polypeptide comprising the amino acid sequence of SEQ ID NO: 696, a polypeptide comprising the amino acid sequence of SEQ ID NO: 697, a polypeptide comprising the amino acid sequence of SEQ ID NO: 698, a polypeptide comprising the amino acid sequence of SEQ ID NO: 699, a polypeptide comprising the amino acid sequence of SEQ ID NO: 700, a polypeptide comprising the amino acid sequence of SEQ ID NO: 701, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 702.

[0182] In some embodiments, the antigen binding molecule according to the present disclosure is capable of binding to ACE2: a polypeptide comprising the amino acid sequence of SEQ ID NO: 8, a polypeptide comprising the amino acid sequence of SEQ ID NO: 703, a polypeptide comprising the amino acid sequence of SEQ ID NO: 704, a polypeptide comprising the amino acid sequence of SEQ ID NO: 705, a polypeptide comprising the amino acid sequence of SEQ ID NO: 706, a polypeptide comprising the amino acid sequence of SEQ ID NO: 707, a polypeptide comprising the amino acid sequence of SEQ ID NO: 708, a polypeptide comprising the amino acid sequence of SEQ ID NO: 709, a polypeptide comprising the amino acid sequence of SEQ ID NO: 710, a polypeptide comprising the amino acid sequence of SEQ ID NO: 711, a polypeptide comprising the amino acid sequence of SEQ ID NO: 712 a polypeptide comprising the amino acid sequence of SEQ ID NO: 713, a polypeptide comprising the amino acid sequence of SEQ ID NO: 714, a polypeptide comprising the amino acid sequence of SEQ ID NO: 715, a polypeptide comprising the amino acid sequence of SEQ ID NO: 716, a polypeptide comprising the amino acid sequence of SEQ ID NO: 717, a polypeptide comprising the amino acid sequence of SEQ ID NO: 718, and a polypeptide comprising the amino acid sequence of SEQ ID NO: 719.

[0183] In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 11; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 12; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 13; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 14; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 15; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 16; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 17.In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 11; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 12; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 13; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 14; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 15; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 16; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 17; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 18; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 19.In a preferred embodiment, the antigen-binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 1; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 11; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 12; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 13; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 14; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 15; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 16. inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 17; inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 18; inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 19; inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 20; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 21; and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 26, and inhibits the interaction between ACE2 and a polypeptide comprising or consisting of the amino acid sequence of SEQ ID NO: 27.

[0184] In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 7; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 686; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 687; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 688; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 689; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 690; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 691; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 692. In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 7; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 686; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 687; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 688; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 689; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 690; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 691; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 692; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 693; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 694.In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 7; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 686; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 687; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 688; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 689; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 690; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 691. inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 692; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 693; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 694; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 695; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 696; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 697 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 698.

[0185] In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 8; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 703; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 704; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 705; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 706; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 707; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 708; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 709. In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 8; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 703; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 704; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 705; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 706; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 707; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 708; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 709; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 710; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 711.In a preferred embodiment, the antigen binding molecule according to the present disclosure inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 8; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 703; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 704; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 705; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 706; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 707; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 708. inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 709; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 710; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 711; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 712; inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 713; and inhibits the interaction between ACE2 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 714 and a polypeptide comprising the amino acid sequence of SEQ ID NO: 715.

[0186] The ability of a given antigen-binding molecule to inhibit the interaction between the SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein and ACE2 can be determined, for example, by analyzing the interaction with the antigen-binding molecule in the presence of one or both interaction partners or after incubation with one or both interaction partners. Antigen-binding molecules that inhibit the interaction between the SARS-CoV-2 spike protein / a given SARS-CoV-2 variant spike protein and ACE2 are identified by observing a reduced / diminished level of interaction between the interaction partner and the antigen-binding molecule in the presence of the interaction partner or after incubation with the interaction partner, compared to the level of interaction observed in the absence of the antigen-binding molecule (or in the presence of a suitable control antigen-binding molecule known not to affect the interaction between the SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein and ACE2). Suitable analyses can be performed in vitro, for example, using recombinant interaction partners or using cells expressing the interaction partner. Cells expressing the interaction partner may express it endogenously or from a nucleic acid introduced into the cell. For the purposes of such assays, one or both of the interaction partners and / or the antigen-binding molecule may be labeled or used with a detectable entity for the purpose of detecting and / or measuring the level of interaction.

[0187] The ability of a given antigen-binding molecule to inhibit the interaction between ACE2 and a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) can be assessed in a pseudovirus neutralization assay, which uses, for example, a vesicular stomatitis virus (VSV) or retrovirus (RV) vector pseudotyped with the SARS-CoV-2 spike protein or a SARS-CoV-2 variant spike protein. Pseudovirus neutralization assays that can be used to evaluate the ability of a given antigen-binding molecule to inhibit the interaction between ACE2 and a sarbecovirus spike protein (e.g., a SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) are described, for example, in Donofrio et al., Vaccines (Basel) (2021) 9(4):389, Nie et al., Emerg. Microbes Infect. (2020) 9:680-686, Chia et al., Sci Adv. (2023) 9(30):eade3470, and Tan et al., Nature Biotechnology (2020) 38:1073-1078, all of which are incorporated by reference in their entireties.

[0188] The ability of a given antigen-binding molecule to inhibit the interaction between ACE2 and a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) can also be assessed in a surrogate virus neutralization test (sVNT), which investigates the binding of the SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein (or a domain thereof, e.g., its RBD) to ACE2 using a labeled species in an ELISA-based assay to infer inhibition of the interaction. Surrogate virus neutralization tests that can be used to evaluate the ability of a given antigen-binding molecule to inhibit the interaction between ACE2 and a sarbecovirus spike protein (e.g., a SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) are described, for example, in Chia et al., Sci Adv. (2023) 9(30):eade3470, Tan et al., Nature Biotechnology (2020) 38:1073-1078, and Springer et al., Diagnostics (Basel). (2023) 13(13):2278, which are incorporated by reference in their entireties.

[0189] In some embodiments, the ability of an antigen binding molecule to inhibit the interaction between ACE2 and a sarbecovirus spike protein (e.g., a SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) is analyzed essentially as described in Example 1.2 herein. In some embodiments, the ability of an antigen binding molecule to inhibit the interaction between ACE2 and a sarbecovirus spike protein (e.g., a SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) is analyzed essentially as described in Example 1.3 herein.

[0190] In some embodiments, an antigen binding molecule according to the present disclosure inhibits the interaction between a SARS-CoV-2 spike protein / a given SARS-CoV-2 variant spike protein and ACE2, as compared to the SARS-CoV-2 spike protein observed in the absence of the antigen binding molecule (or in the presence of a suitable control antigen binding molecule known not to affect the interaction between a SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein and ACE2). or (b) reduce / inhibit the level of interaction between the SARS-CoV-2 variant spike protein and ACE2 by less than 1-fold, for example, <0.99-fold, <0.95-fold, <0.9-fold, <0.85-fold, <0.8-fold, <0.75-fold, <0.7-fold, <0.65-fold, <0.6-fold, <0.55-fold, <0.5-fold, <0.45-fold, <0.4-fold, <0.35-fold, <0.3-fold, <0.25-fold, <0.2-fold, <0.15-fold, <0.1-fold, <0.05-fold or <0.01-fold.

[0191] In some embodiments, the antigen binding molecule inhibits the interaction between the SARS-CoV-2 spike protein / given SARS-CoV-2 variant spike protein and ACE2 at a neutralization level of less than 1 μg / ml, preferably less than 800 ng / ml, less than 700 ng / ml, less than 600 ng / ml, less than 500 ng / ml, less than 400 ng / ml, less than 300 ng / ml, less than 200 ng / ml, less than 100 ng / ml, less than 90 ng / ml, less than 80 ng / ml, as determined, for example, in a pseudovirus neutralization assay performed as described in Example 1.2 herein. ng / ml, ≦70ng / ml, ≦60ng / ml, ≦50ng / ml, ≦40ng / ml, ≦30ng / ml, ≦20ng / ml, ≦10ng / ml, ≦9ng / ml, ≦8ng / ml, ≦7ng / ml, ≦6ng / ml, ≦5ng / ml, ≦4ng / ml, ≦3n g / ml, ≦2ng / ml, ≦1ng / ml, ≦900pg / ml, ≦800pg / ml, ≦700pg / ml, ≦600pg / ml , ≦500pg / ml, ≦400pg / ml, ≦300pg / ml, ≦200pg / ml or ≦100pg / ml. 50In some embodiments, the antigen binding molecule can inhibit the interaction between the SARS-CoV-2 spike protein / given SARS-CoV-2 variant spike protein and ACE2 at a neutralization level of less than 1 μg / ml, preferably less than 800 ng / ml, less than 700 ng / ml, less than 600 ng / ml, less than 500 ng / ml, less than 400 ng / ml, less than 300 ng / ml, less than 200 ng / ml, less than 100 ng / ml, less than 90 ng / ml, less than 80 ng / ml, ≦70ng / ml, ≦60ng / ml, ≦50ng / ml, ≦40ng / ml, ≦30ng / ml, ≦20ng / ml, ≦10ng / ml, ≦9ng / ml, ≦8ng / ml, ≦7ng / ml, ≦6ng / ml, ≦5ng / ml, ≦4ng / ml, ≦3n g / ml, ≦2ng / ml, ≦1ng / ml, ≦900pg / ml, ≦800pg / ml, ≦700pg / ml, ≦600pg / ml , ≦500pg / ml, ≦400pg / ml, ≦300pg / ml, ≦200pg / ml or ≦100pg / ml. 50 can be inhibited by

[0192] In some embodiments, an antigen binding molecule according to the present disclosure reduces / inhibits infection of ACE2-expressing cells by a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 and / or one or more SARS-CoV-2 variants). Such antigen binding molecules may be described as inhibiting / antagonizing infection of ACE2-expressing cells or may be referred to as neutralizing infection of such cells by a sarbecovirus(es).

[0193] In some embodiments, an antigen binding molecule according to the present disclosure reduces / inhibits infection of ACE2-expressing cells by a SARS-CoV-2 variant selected from BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, XBB, XBB.1, XBB.1.16, XBB.2.3, EG.5, EG.5.1, B.1.1.7, B.1.351, B.1.617.2 and P.1.

[0194] In some embodiments, an antigen binding molecule according to the present disclosure can (independently) inhibit infection of ACE2-expressing cells by two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15 or more) SARSr-CoVs selected from SARS-CoV-2 and SARS-CoV-2 variants. That is, in some embodiments, an antigen binding molecule that inhibits infection of ACE2-expressing cells by a given (first) SARSr-CoV selected from SARS-CoV-2 and SARS-CoV-2 variants also inhibits infection of ACE2-expressing cells by one or more additional (second, third, etc.) SARSr-CoVs selected from SARS-CoV-2 and SARS-CoV-2 variants, where the one or more additional SARSr-CoVs have a nucleotide sequence that differs from that of the first SARSr-CoV.

[0195] In some embodiments, an antigen binding molecule according to the present disclosure inhibits infection of ACE2-expressing cells by two or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, or all 18) SARSr-CoVs selected from SARS-CoV-2, BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, XBB, XBB.1, XBB.1.16, XBB.2.3, EG.5, EG.5.1, B.1.1.7, B.1.351, B.1.617.2, and P.1.

[0196] In a preferred embodiment, an antigen binding molecule according to the present disclosure inhibits infection of ACE2-expressing cells by SARS-CoV-2, BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, and BF.7. In a preferred embodiment, an antigen binding molecule according to the present disclosure inhibits infection of ACE2-expressing cells by SARS-CoV-2, BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, and XBB.1. In a preferred embodiment, the antigen binding molecules according to the present disclosure inhibit infection of ACE2-expressing cells by SARS-CoV-2, BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, XBB.1, XBB.1.16, XBB.2.3, EG.5 and EG.5.1.

[0197] The ability of a given antigen-binding molecule to inhibit infection of ACE2-expressing cells by a SARS-CoV-2 / SARS-CoV-2 variant can be analyzed by detecting / quantifying infection of ACE2-expressing cells by the SARS-CoV-2 / SARS-CoV-2 variant (or a pseudovirus pseudotyped with the spike protein encoded by the SARS-CoV-2 / SARS-CoV-2 variant) in the presence of the antigen-binding molecule and comparing the level of infection to that observed in the absence of the antigen-binding molecule (and / or the level of infection observed in the presence of a suitable control antigen-binding molecule known not to affect infection of ACE2-expressing cells by the relevant virus). Such methods can include determining the absolute number of, or the percentage of, cells infected with (e.g., containing) the relevant virus.

[0198] The ability of a given antigen-binding molecule to inhibit infection of ACE2-expressing cells by SARS-CoV-2 / SARS-CoV-2 variants can be analyzed by pseudovirus neutralization assays, for example, as described in Chia et al., Sci Adv. (2023) 9(30):eade3470 or Tan et al., Nature Biotechnology (2020) 38:1073-1078.

[0199] In some embodiments, the antigen binding molecules of the present disclosure inhibit infection of ACE2-expressing cells by SARS-CoV-2 / SARS-CoV-2 variants (or pseudoviruses pseudotyped with spike proteins encoded by SARS-CoV-2 / SARS-CoV-2 variants) in the absence of the antigen binding molecule (or inhibit infection of ACE2-expressing cells by SARS-CoV-2 / SARS-CoV-2 variants). the level of infection of ACE2-expressing cells observed in a control antibody (in the presence of a suitable control antigen-binding molecule known to be non-specific) is reduced / inhibited by less than 1-fold, e.g., ≦0.99-fold, ≦0.95-fold, ≦0.9-fold, ≦0.85-fold, ≦0.8-fold, ≦0.75-fold, ≦0.7-fold, ≦0.65-fold, ≦0.6-fold, ≦0.55-fold, ≦0.5-fold, ≦0.45-fold, ≦0.4-fold, ≦0.35-fold, ≦0.3-fold, ≦0.25-fold, ≦0.2-fold, ≦0.15-fold, ≦0.1-fold, ≦0.05-fold or ≦0.01-fold.

[0200] In some embodiments, the antigen binding molecule inhibits infection of ACE2-expressing cells by a SARS-CoV-2 / SARS-CoV-2 variant (or a pseudovirus pseudotyped with a spike protein encoded by a SARS-CoV-2 / SARS-CoV-2 variant) at a neutralization level of less than 1 μg / ml, preferably ≦800 ng / ml, ≦700 ng / ml, ≦600 ng / ml, ≦500 ng / ml, ≦400 ng / ml, ≦300 ng / ml, ≦200 ng / ml, as determined, for example, in a pseudovirus neutralization assay performed as described in Example 1.2 herein. g / ml, ≦100ng / ml, ≦90ng / ml, ≦80ng / ml, ≦70ng / ml, ≦60ng / ml, ≦50ng / ml, ≦40ng / m l, ≦30ng / ml, ≦20ng / ml, ≦10ng / ml, ≦9ng / ml, ≦8ng / ml, ≦7ng / ml, ≦6ng / ml, ≦5ng / m l, ≦4ng / ml, ≦3ng / ml, ≦2ng / ml, ≦1ng / ml, ≦900pg / ml, ≦800pg / ml, ≦700pg / ml, ≦60 IC of one of 0pg / ml, ≦500pg / ml, ≦400pg / ml, ≦300pg / ml, ≦200pg / ml or ≦100pg / ml 50 can be inhibited by

[0201] In some embodiments, antigen binding molecules according to the present disclosure have one or more novel, similar, or improved functional properties compared to known antigen binding molecules that bind to SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins.

[0202] In some embodiments, the antigen-binding molecule has one or more novel, similar, or improved functional properties compared to, for example, SS6V11-E7 (also referred to herein as "E7") described in WO 2022 / 245288 A1. For purposes of comparing functional properties in the following paragraphs, "SS6V11-E7" refers to an antigen-binding molecule formed by the association of two polypeptides consisting of SEQ ID NO: 837 and two polypeptides consisting of SEQ ID NO: 838.

[0203] In some embodiments, the antigen-binding molecule has one or more novel, similar, or improved functional properties compared to LyCov-1404 (also known as bebtelovimab; DrugBank accession number DB16755). For purposes of comparison of functional properties in the following paragraphs, "LyCov-1404" refers to the antigen-binding molecule formed by the association between two polypeptides consisting of SEQ ID NO: 854 and two polypeptides consisting of SEQ ID NO: 855.

[0204] In some embodiments, the antigen-binding molecules described herein may exhibit one or more of the following: binds to SARS-CoV-2 variant spike proteins that SS6V11-E7 and / or LyCov-1404 do not bind; inhibiting the interaction between ACE2 and SARS-CoV-2 variant spike proteins whose interaction with ACE2 is not inhibited by SS6V11-E7 and / or LyCov-1404; inhibits infection of ACE2-expressing cells by sarbecoviruses (e.g., SARS-CoV-2 variants) whose infection of ACE2-expressing cells is not inhibited by SS6V11-E7 and / or LyCov-1404; Similar or increased affinity (e.g., similar or lower K) compared to the affinity with which the related protein(s) are bound by SS6V11-E7 and / or LyCov-1404. D ) and binds to a sarbecovirus spike protein (e.g., a SARSr-CoV spike protein; e.g., a SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins). inhibits the interaction between a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) and ACE2 with similar or increased potency (e.g., similar or lower IC50) compared to the potency with which such interaction is inhibited by SS6V11-E7 and / or LyCov-1404; and / or Inhibits infection of ACE2-expressing cells by a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 and / or one or more SARS-CoV-2 variants) with similar or increased potency (e.g., similar or lower IC50) compared to the potency with which such infection is inhibited by SS6V11-E7 and / or LyCov-1404.

[0205] Consistent with the preceding paragraph, Criterion K D / K that is "similar" to IC50 values D / IC50 value is the standard K D / IC50 value, e.g., ≧0.55-fold and ≦1.9-fold, ≧0.6-fold and ≦1.8-fold, ≧0.65-fold and ≦1.7-fold, ≧0.7-fold and ≦1.6-fold, ≧0.75-fold and ≦1.5-fold, ≧0.8-fold and ≦1.4-fold, ≧0.85-fold and ≦1.3-fold, ≧0.9-fold and ≦1.2-fold, or ≧0.95-fold and ≦1.1-fold. D / lower K compared to IC50 value D / IC50 value is K D / It may be less than 1-fold the IC50 value, for example, ≦0.99-fold, ≦0.95-fold, ≦0.9-fold, ≦0.85-fold, ≦0.8-fold, ≦0.75-fold, ≦0.7-fold, ≦0.65-fold, ≦0.6-fold, ≦0.55-fold, ≦0.5-fold, ≦0.45-fold, ≦0.4-fold, ≦0.35-fold, ≦0.3-fold, ≦0.25-fold, ≦0.2-fold, ≦0.15-fold, ≦0.1-fold, ≦0.05-fold or ≦0.01-fold.

[0206] It will be understood that for the purposes of such evaluation, equal amounts / concentrations of the antigen-binding molecule and SS6V11-E7 and / or LyCov-1404 may be compared. In some embodiments, the antigen binding molecules of the present disclosure bind to the SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein with a KD that is similar to or less than the KD for binding of SS6V11-E7 and / or LyCov-1404 to the relevant protein, determined in the same assay. In some embodiments, the antigen binding molecule of the disclosure binds to a SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein with a KD that is ≥ 0.5-fold and ≤ 2-fold, e.g., one of ≥ 0.55-fold and ≤ 1.9-fold, ≥ 0.6-fold and ≤ 1.8-fold, ≥ 0.65-fold and ≤ 1.7-fold, ≥ 0.7-fold and ≤ 1.6-fold, ≥ 0.75-fold and ≤ 1.5-fold, ≥ 0.8-fold and ≤ 1.4-fold, ≥ 0.85-fold and ≤ 1.3-fold, ≥ 0.9-fold and ≤ 1.2-fold, or ≥ 0.95-fold and ≤ 1.1-fold the KD for binding of SS6V11-E7 and / or LyCov-1404 to the relevant protein, determined in the same assay. In some embodiments, the antigen binding molecule of the disclosure binds to a SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein with a KD that is less than 1-fold the KD for SS6V11-E7 and / or LyCov-1404 binding to the related protein, e.g., <0.99-fold, <0.95-fold, <0.9-fold, <0.85-fold, <0.8-fold, <0.75-fold, <0.7-fold, <0.65-fold, <0.6-fold, <0.55-fold, <0.5-fold, <0.45-fold, <0.4-fold, <0.35-fold, <0.3-fold, <0.25-fold, <0.2-fold, <0.15-fold, <0.1-fold, <0.05-fold, or <0.01-fold, determined in the same assay.

[0207] In some embodiments, the antigen binding molecules of the present disclosure inhibit the interaction between the SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein and ACE2, as determined in the same assay, by the IC when SS6V11-E7 and / or LyCov-1404 inhibit the interaction between the relevant protein and ACE2. 50 IC that is similar to or less than 50 In some embodiments, the antigen binding molecule inhibits the interaction between the SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein and ACE2 with an IC50 value determined in the same assay for inhibition of the interaction between related proteins and ACE2 by SS6V11-E7 and / or LyCov-1404. 50 an IC that is ≥ 0.5-fold and ≤ 2-fold, e.g., ≥ 0.55-fold and ≤ 1.9-fold, ≥ 0.6-fold and ≤ 1.8-fold, ≥ 0.65-fold and ≤ 1.7-fold, ≥ 0.7-fold and ≤ 1.6-fold, ≥ 0.75-fold and ≤ 1.5-fold, ≥ 0.8-fold and ≤ 1.4-fold, ≥ 0.85-fold and ≤ 1.3-fold, ≥ 0.9-fold and ≤ 1.2-fold, or ≥ 0.95-fold and ≤ 1.1-fold 50 In some embodiments, the antigen binding molecule inhibits the interaction between the SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein and ACE2 with an IC50 value determined in the same assay for inhibition of the interaction between related proteins and ACE2 by SS6V11-E7 and / or LyCov-1404. 50 an IC that is less than 1 times the original concentration, for example, ≦0.99 times, ≦0.95 times, ≦0.9 times, ≦0.85 times, ≦0.8 times, ≦0.75 times, ≦0.7 times, ≦0.65 times, ≦0.6 times, ≦0.55 times, ≦0.5 times, ≦0.45 times, ≦0.4 times, ≦0.35 times, ≦0.3 times, ≦0.25 times, ≦0.2 times, ≦0.15 times, ≦0.1 times, ≦0.05 times, or ≦0.01 times the original concentration 50 and inhibits.

[0208] In some embodiments, the antigen binding molecules of the present disclosure inhibit infection of ACE2-expressing cells with SARS-CoV-2 / SARS-CoV-2 variants (or pseudoviruses pseudotyped with spike proteins encoded by SARS-CoV-2 / SARS-CoV-2 variants) by an IC determined in the same assay when SS6V11-E7 and / or LyCov-1404 inhibit the interaction between the relevant protein and ACE2. 50 IC that is similar to or less than 50 In some embodiments, the antigen binding molecule inhibits infection of ACE2-expressing cells by a SARS-CoV-2 / SARS-CoV-2 variant (or a pseudovirus pseudotyped with the spike protein encoded by the SARS-CoV-2 / SARS-CoV-2 variant) at an IC50 or higher for inhibition of infection of such cells by the related SARSr-CoV by SS6V11-E7 and / or LyCov-1404, determined in the same assay. 50 an IC that is ≥ 0.5-fold and ≤ 2-fold, e.g., ≥ 0.55-fold and ≤ 1.9-fold, ≥ 0.6-fold and ≤ 1.8-fold, ≥ 0.65-fold and ≤ 1.7-fold, ≥ 0.7-fold and ≤ 1.6-fold, ≥ 0.75-fold and ≤ 1.5-fold, ≥ 0.8-fold and ≤ 1.4-fold, ≥ 0.85-fold and ≤ 1.3-fold, ≥ 0.9-fold and ≤ 1.2-fold, or ≥ 0.95-fold and ≤ 1.1-fold 50 In some embodiments, the antigen binding molecule inhibits infection of ACE2-expressing cells by a SARS-CoV-2 / SARS-CoV-2 variant (or a pseudovirus pseudotyped with the spike protein encoded by the SARS-CoV-2 / SARS-CoV-2 variant) at an IC50 or higher for inhibition of infection of such cells by the related SARSr-CoV by SS6V11-E7 and / or LyCov-1404, determined in the same assay. 50of the IC, which is less than 1 time, for example, ≦0.99 times, ≦0.95 times, ≦0.9 times, ≦0.85 times, ≦0.8 times, ≦0.75 times, ≦0.7 times, ≦0.65 times, ≦0.6 times, ≦0.55 times, ≦0.5 times, ≦0.45 times, ≦0.4 times, ≦0.35 times, ≦0.3 times, ≦0.25 times, ≦0.2 times, ≦0.15 times, ≦0.1 times, ≦0.05 times, or ≦0.01 times 50 and inhibits.

[0209] Detailed Exemplary Antigen-Binding Molecules and Polypeptides The present disclosure also provides polypeptide compositions of antigen-binding molecules. The polypeptides may be provided in isolated or substantially purified form.

[0210] The antigen-binding molecules of the present disclosure may be or may comprise a complex of polypeptides. As used herein, it will be understood that when a polypeptide comprises more than one domain or region, the multiple domains / regions are preferably present on the same polypeptide chain, i.e., a polypeptide comprising more than one domain or region is a fusion polypeptide comprising the domains / regions.

[0211] In some embodiments, a polypeptide according to the present disclosure comprises or consists of a VH described herein. In some embodiments, a polypeptide according to the present disclosure comprises or consists of a VL described herein.

[0212] In some embodiments, the polypeptide further comprises one or more antibody heavy chain constant regions (CH). In some embodiments, the polypeptide further comprises one or more antibody light chain constant regions (CL). In some embodiments, the polypeptide comprises an immunoglobulin (Ig) CH1, CH2, and / or CH3 region.

[0213] In some embodiments, the polypeptide comprises one or more regions of an immunoglobulin heavy chain constant sequence. In some embodiments, the polypeptide comprises a CH1 region described herein. In some embodiments, the polypeptide comprises a hinge region described herein. In some embodiments, the polypeptide comprises a CH2 region described herein. In some embodiments, the polypeptide comprises a CH3 region described herein. In some embodiments, the polypeptide comprises a CH2-CH3 region described herein. In some embodiments, the polypeptide comprises a CH1-hinge-CH2-CH3 region described herein.

[0214] In some embodiments, the polypeptide comprises one or more regions of an immunoglobulin light chain constant sequence, hi some embodiments, the polypeptide comprises a CL region described herein.

[0215] In some embodiments, the polypeptide according to the present disclosure is: (i) VH (ii) VL (iii) VH-CH1 (iv) VL-CL (v) VL-CH1 (vi) VH-CL (vii) VH-CH1-CH2-CH3 (viii) VL-CL-CH2-CH3 (ix) VL-CH1-CH2-CH3 (x)VH-CL-CH2-CH3 The structure from N-terminus to C-terminus is one of:

[0216] The present disclosure also provides antigen-binding molecules composed of the polypeptides of the present disclosure. In some embodiments, the antigen-binding molecules of the present disclosure comprise the following combinations of polypeptides: (A) VH+VL (B) VH-CH1+VL-CL (C) VL-CH1+VH-CL (D) VH-CH1-CH2-CH3+VL-CL (E) VH-CL-CH2-CH3+VL-CH1 (F) VL-CH1-CH2-CH3+VH-CL (G)VL-CL-CH2-CH3+VH-CH1 (H)VH-CH1-CH2-CH3+VL-CL-CH2-CH3 (I)VH-CL-CH2-CH3+VL-CH1-CH2-CH3 Contains one of the following:

[0217] In some embodiments, the antigen-binding molecule comprises more than one of the polypeptide combinations shown in (A) to (I) above. By way of example, with reference to (D) above, in some embodiments, the antigen-binding molecule comprises two polypeptides comprising the structure VH-CH1-CH2-CH3 and two polypeptides comprising the structure VL-CL.

[0218] Consistent with (i) through (x) and (A) through (I) above, "VH" refers to a VH region as described herein, and "VL" refers to a VL region as described herein. In some embodiments, the antigen binding molecule of the present disclosure is selected from the group consisting of SEQ ID NOs: 36, 52, 67, 83, 96, 105, 120, 136, 149, 164, 179, 193, 206, 220, 235, 249, 262, 274, 285, 299, 312, 325, 336, 350, 362, 368, 381, 393, 405, 416, 427, 436, 449, 453, 464, 475, 487, 496, 508, 522, 535, 547, 559, 57 2, 584, 592, 600, 614, 627, 643, or 656.

[0219] In some embodiments, the antigen binding molecule of the present disclosure is selected from the group consisting of SEQ ID NOs: 44, 59, 75, 89, 102, 113, 128, 144, 157, 171, 187, 200, 213, 228, 242, 256, 270, 280, 291, 305, 318, 331, 343, 355, 366, 374, 388, 400, 411, 423, 432, 443, 451, 460, 471, 481, 491, 502, 515, 529, 542, 554, 567, 579, 587, 596, 607, 621, 635, 651, 663, or a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) amino acid sequence identity to 579, 587, 596, 607, 621, 635, 651, or 663.

[0220] In some embodiments, the antigen binding molecule of the present disclosure is selected from the group consisting of SEQ ID NOs: 720, 722, 724, 726, 728, 730, 732, 734, 736, 738, 740, 742, 744, 746, 748, 750, 752, 754, 756, 758, 760, 762, 764, 766, 768, 770, 772, 774, 776, 778, 780, 782, 784, 786, 788, 790, 792, 794, 796, 798, 800, 802, 804 , 806, 808, 810, 812, 814, 816, 818, or 820.

[0221] In some embodiments, the antigen binding molecule of the present disclosure is selected from the group consisting of SEQ ID NOs: 721, 723, 725, 727, 729, 731, 733, 735, 737, 739, 741, 743, 745, 747, 749, 751, 753, 755, 757, 759, 761, 763, 765, 767, 769, 771, 773, 775, 777, 779, 781, 783, 785, 787, 789, 791, 793, 795, 797, 799, 801, 803, 805 , 807, 809, 811, 813, 815, 817, 819 or 821.

[0222] In some embodiments, antigen-binding molecules of the present disclosure comprise one or more polypeptides comprising a VH region comprising the heavy chain CDRs and a VL region comprising the light chain CDRs of an antibody selected from the antibodies shown in Table A herein. That is, in some embodiments, antigen-binding molecules comprise one or more polypeptides comprising (i) a VH region comprising the HC-CDR1, HC-CDR2, and HC-CDR3 shown in column A of Table A, and (ii) a VL region comprising the LC-CDR1, LC-CDR2, and LC-CDR3 shown in column B of Table A, wherein the sequences in columns A and B are selected from the same row of Table A. In some embodiments, antigen-binding molecules of the present disclosure comprise one or more polypeptides comprising a VH region comprising the heavy chain CDRs and a VL region comprising the light chain CDRs of an antibody shown in Table A herein. In some embodiments, antigen-binding molecules of the present disclosure comprise one or more polypeptides comprising a VH region comprising the heavy chain CDRs and a VL region comprising the light chain CDRs of an antibody shown in Table A herein.

[0223] In some embodiments, antigen-binding molecules of the present disclosure comprise one or more polypeptides comprising a VH region comprising heavy chain FRs and a VL region comprising light chain FRs of an antibody selected from the antibodies shown in Table B herein. That is, in some embodiments, antigen-binding molecules comprise one or more polypeptides comprising (i) a VH region comprising HC-FR1, HC-FR2, HC-FR3, and HC-FR4 shown in column A of Table B, and (ii) a VL region comprising LC-FR1, LC-FR2, LC-FR3, and LC-FR4 shown in column B of Table B, wherein the sequences in columns A and B are selected from the same row of Table B. In some embodiments, antigen-binding molecules of the present disclosure comprise one or more polypeptides comprising a VH region comprising heavy chain FRs and a VL region comprising light chain FRs of an antibody shown in Table B herein. In some embodiments, antigen-binding molecules of the present disclosure comprise one or more polypeptides comprising a VH region comprising heavy chain FRs and a VL region comprising light chain FRs of an antibody shown in Table B herein.

[0224] In some embodiments, an antigen-binding molecule of the disclosure comprises (i) an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) amino acid sequence identity to the amino acid sequence shown in column A of Table C, and (ii) an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99%, or 100%) amino acid sequence identity to the amino acid sequence shown in column B of Table C. and wherein the sequences in columns A and B are selected from the same row of Table C.

[0225] In some embodiments, an antigen-binding molecule of the present disclosure comprises one or more polypeptides comprising the VH and VL regions of an antibody clone selected from the antibodies shown in Table C herein. That is, in some embodiments, an antigen-binding molecule comprises one or more polypeptides comprising (i) the amino acid sequence shown in column A of Table C, and (ii) the amino acid sequence shown in column B of Table C, wherein the sequences in columns A and B are selected from the same row of Table C. In some embodiments, an antigen-binding molecule of the present disclosure comprises one or more polypeptides comprising the VH and VL regions of an antibody shown in Table C herein. In some embodiments, an antigen-binding molecule of the present disclosure comprises one or more polypeptides comprising the VH and VL regions of an antibody shown in Table C herein.

[0226] In some embodiments, an antigen-binding molecule of the present disclosure is (i) a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≧70%, ≧75%, ≧80%, ≧85%, ≧86%, ≧87%, ≧88%, ≧89%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99% or 100%) amino acid sequence identity to an amino acid sequence set forth in column A of Table D, and (ii) and polypeptides comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to the amino acid sequence shown in column B of Table D, wherein the sequences in columns A and B are selected from the same row of Table D.

[0227] In some embodiments, an antigen-binding molecule of the present disclosure comprises a polypeptide of an antigen-binding molecule according to Table D herein. That is, in some embodiments, the antigen-binding molecule comprises (i) a polypeptide comprising or consisting of the amino acid sequence set forth in column A of Table D, and (ii) a polypeptide comprising or consisting of the amino acid sequence set forth in column B of Table D, wherein the sequences in columns A and B are selected from the same row of Table D.

[0228] In some embodiments, the antigen binding molecule of the present disclosure comprises: (1) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 722, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 723; (2) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 720, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 721; (3) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 724, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 725; (4) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 726, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 727; (5) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 794, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 795; (6) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 752, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 753; (7) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 748, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 749; (8) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 740, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 741; (9) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 754, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 754; (10) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 766, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 767; (11) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 774, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 775; (12) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 776, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 777; (13) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 778, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 779; (14) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 780, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 781; (15) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 784, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 785; (16) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 786, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 787; (17) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 728, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 729; (18) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 790, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 791; (19) (i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 806, and (ii) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 807; Includes.

[0229] Known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecules Aspects and embodiments of the present disclosure also relate to known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecules and derivatives thereof. For clarity, when referring to "antigen-binding molecules of the present disclosure" herein, it is not intended to refer to such known antigen-binding molecules.

[0230] In some embodiments, the known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule is SS6V11-E7 or a derivative thereof. SS6V11-E7 (also referred to herein as "E7") is described, for example, in WO2022 / 245288A1. E7 comprises a VH region set forth in SEQ ID NO: 824 and a VL region set forth in SEQ ID NO: 830. The HC-CDR1, HC-CDR2, and HC-CDR3 of E7 are set forth in SEQ ID NOs: 825, 826, and 827 (respectively), and the LC-CDR1, LC-CDR2, and LC-CDR3 of E7 are set forth in SEQ ID NOs: 831, 832, and 832 (respectively). HC-FR1, HC-FR2, HC-FR3, and HC-FR4 of E7 are set forth in SEQ ID NOs: 266, 828, 829, and 112 (respectively), and LC-FR1, LC-FR2, LC-FR3, and LC-FR4 of E7 are set forth in SEQ ID NOs: 834, 835, 826, and 486 (respectively). E7, in its human IgG1 heavy chain, kappa light chain format, is formed by association between two polypeptides having the sequence of SEQ ID NO: 837 and two polypeptides having the sequence of SEQ ID NO: 838.

[0231] In some embodiments, SS6V11-E7 or a derivative thereof comprises (i) HC-CDR1=SEQ ID NO: 825 (or a variant thereof in which one, two, or three amino acids in HC-CDR1 are substituted with another amino acid), HC-CDR2=SEQ ID NO: 826 (or a variant thereof in which one, two, or three amino acids in HC-CDR2 are substituted with another amino acid), and HC-CDR3=SEQ ID NO: 827 (or a variant thereof in which one, two, or three amino acids in HC-CDR3 are substituted with another amino acid). and (ii) a VL region comprising LC-CDR1=SEQ ID NO: 833 (or a variant thereof in which one, two or three amino acids in LC-CDR1 are substituted with another amino acid), LC-CDR2=SEQ ID NO: 832 (or a variant thereof in which one, two or three amino acids in LC-CDR2 are substituted with another amino acid), and LC-CDR3=SEQ ID NO: 833 (or a variant thereof in which one, two or three amino acids in LC-CDR3 are substituted with another amino acid). In some embodiments, SS6V11-E7 or a derivative thereof comprises: (i) a VH region having at least 70%, preferably ≧80%, ≧85%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99% or 100% amino acid sequence identity to SEQ ID NO: 824; and (ii) a VL region having at least 70%, preferably ≧80%, ≧85%, ≧90%, ≧91%, ≧92%, ≧93%, ≧94%, ≧95%, ≧96%, ≧97%, ≧98%, ≧99% or 100% amino acid sequence identity to SEQ ID NO: 830.

[0232] In some embodiments, the known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule is LyCov-1404 or a derivative thereof. LyCov-1404 (also known as bebterovimab; DrugBank accession number DB16755) comprises a VH region set forth in SEQ ID NO: 839 and a VL region set forth in SEQ ID NO: 846. The HC-CDR1, HC-CDR2, and HC-CDR3 of LyCov-1404 are set forth in SEQ ID NOs: 840, 841, and 842 (respectively), and the LC-CDR1, LC-CDR2, and LC-CDR3 of LyCov-1404 are set forth in SEQ ID NOs: 847, 848, and 849 (respectively). HC-FR1, HC-FR2, HC-FR3 and HC-FR4 of LyCov-1404 are set forth in SEQ ID NOs: 843, 844, 845 and 112 (respectively), and LC-FR1, LC-FR2, LC-FR3 and LC-FR4 of LyCov-1404 are set forth in SEQ ID NOs: 850, 851, 852 and 853 (respectively). LyCov-1404 in a human IgG1 (G1m3) heavy chain, CλCL2 light chain format is formed by the association between two polypeptides having the sequences of SEQ ID NO: 854 and two polypeptides having the sequences of SEQ ID NO: 855.

[0233] In some embodiments, LyCov-1404 or a derivative thereof comprises (i) HC-CDR1 = SEQ ID NO: 840 (or a variant thereof in which one, two or three amino acids in HC-CDR1 are substituted with another amino acid), HC-CDR2 = SEQ ID NO: 841 (or a variant thereof in which one, two or three amino acids in HC-CDR2 are substituted with another amino acid), and HC-CDR3 = SEQ ID NO: 842 (or a variant thereof in which one, two or three amino acids in HC-CDR3 are substituted with another amino acid). and (ii) a VH region comprising LC-CDR1=SEQ ID NO: 847 (or a variant thereof in which one, two or three amino acids in LC-CDR1 are substituted with another amino acid), LC-CDR2=SEQ ID NO: 848 (or a variant thereof in which one, two or three amino acids in LC-CDR2 are substituted with another amino acid), and LC-CDR3=SEQ ID NO: 849 (or a variant thereof in which one, two or three amino acids in LC-CDR3 are substituted with another amino acid). In some embodiments, LyCov-1404 or a derivative thereof comprises: (i) a VH region having at least 70%, preferably at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% amino acid sequence identity to SEQ ID NO: 839; and (ii) a VL region having at least 70%, preferably at least 80%, at least 85%, at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99% or 100% amino acid sequence identity to SEQ ID NO: 846.

[0234] In some embodiments, the known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein binding antigen binding molecule is (A)(i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% (e.g., one of: ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 824; and (ii) a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 830; (B)(i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% (e.g., one of: ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 839; and (ii) a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 846; (C)(i) a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 837, and and (ii) a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 838; or (D)(i) a polypeptide comprising, or consisting of, an amino acid sequence having at least 70% (e.g., one of: ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 854; and (ii) a polypeptide comprising or consisting of an amino acid sequence having at least 70% (e.g., one of ≥70%, ≥75%, ≥80%, ≥85%, ≥86%, ≥87%, ≥88%, ≥89%, ≥90%, ≥91%, ≥92%, ≥93%, ≥94%, ≥95%, ≥96%, ≥97%, ≥98%, ≥99% or 100%) amino acid sequence identity to SEQ ID NO: 855. Includes.

[0235] Linkers and Additional Sequences The antigen-binding molecules and polypeptides of the present disclosure may additionally comprise additional amino acids or sequences of amino acids.

[0236] The antigen-binding molecules and polypeptides of the present disclosure may contain one or more linker sequences between amino acid sequences. For example, a linker sequence may be provided between the VH sequence and the VL sequence to provide a link between the VH and VL (e.g., as in an scFv molecule).

[0237] Linker sequences are known to those skilled in the art, and are described, for example, in Chen et al., Adv Drug Deliv Rev (2013) 65(10):1357-1369, the entire contents of which are incorporated herein by reference. In some embodiments, the linker sequence may be a flexible linker sequence. A flexible linker sequence allows the relative movement of the amino acid sequences connected by the linker sequence. Flexible linkers are known to those skilled in the art, and some are identified in Chen et al., Adv Drug Deliv Rev (2013) 65(10):1357-1369. Flexible linker sequences often contain a high proportion of glycine and / or serine residues.

[0238] In some embodiments, the linker sequence comprises at least one glycine residue and / or at least one serine residue. In some embodiments, the linker sequence comprises or consists of glycine and serine residues. In some embodiments, the linker sequence has the structure: (GxS)n or (GxS)nGm; where G=glycine, S=serine, x=3 or 4, n=2, 3, 4, 5 or 6, and m=0, 1, 2 or 3. In some embodiments, the linker sequence comprises one or more (e.g., 1, 2, 3, 4, 5, or 6) copies (e.g., in tandem) of the sequence motif G4S. In some embodiments, the linker sequence comprises or consists of (G4S)4 or (G4S)6. In some embodiments, the linker sequence has a length of 1 to 2, 1 to 3, 1 to 4, 1 to 5, 1 to 10, 1 to 15, 1 to 20, 1 to 25, or 1 to 30 amino acids.

[0239] The antigen-binding molecules and polypeptides of the present disclosure may comprise an amino acid sequence(s) that facilitates expression, folding, transport, processing, purification, or detection of the antigen-binding molecule / polypeptide. For example, the antigen-binding molecules and polypeptides of the present disclosure may additionally comprise a sequence of amino acids that forms a detectable moiety, for example, as described herein below.

[0240] The antigen-binding molecules and polypeptides of the present disclosure may additionally contain a signal peptide (also known as a leader sequence or signal sequence). Signal peptides usually consist of a sequence of 5 to 30 hydrophobic amino acids and form a single alpha helix. Secreted proteins and proteins expressed on the cell surface often contain signal peptides. Signal peptides are known for many proteins and are recorded in databases such as GenBank, UniProt, and Ensembl, and / or can be identified / predicted using amino acid sequence analysis tools such as SignalP (Petersen et al., 2011 Nature Methods 8:785-786) or Signal-BLAST (Frank and Sippl, 2008 Bioinformatics 24:2172-2176).

[0241] A signal peptide may be present at the N-terminus of an antigen-binding molecule / polypeptide or may be present in a newly synthesized antigen-binding molecule / polypeptide. The signal peptide provides efficient transport of the antigen-binding molecule / polypeptide. The signal peptide is often removed by cleavage and is therefore not included in the mature antigen-binding molecule / polypeptide.

[0242] Signal peptides are known for many proteins and are recorded in databases such as GenBank, UniProt, Swiss-Prot, TrEMBL, Protein Information Resource, Protein Data Bank, Ensembl and InterPro, and / or can be identified / predicted using amino acid sequence analysis tools such as, for example, SignalP (Petersen et al., 2011 Nature Methods 8:785-786) or Signal-BLAST (Frank and Sippl, 2008 Bioinformatics 24:2172-2176).

[0243] Labels and conjugates In some embodiments, an antigen-binding molecule or polypeptide of the present disclosure comprises a detectable moiety.

[0244] In some embodiments, the detectable moiety is a fluorescent label, a phosphorescent label, a luminescent label, an immunodetectable label (e.g., an epitope tag), a radiolabel, a chemical, a nucleic acid, or an enzymatic label. The antigen-binding molecule or polypeptide can be covalently or non-covalently labeled with the detectable moiety.

[0245] Fluorescent labels include, for example, fluorescein, rhodamine, allophycocyanin, eosin, and NDB, green fluorescent protein (GFP), rare earth chelates such as europium (Eu), terbium (Tb), and samarium (Sm), tetramethylrhodamine, Texas Red, 4-methylumbelliferone, 7-amino-4-methylcoumarin, Cy3, and Cy5. Radiolabels include, for example, hydrogen 3 , sulfur 35 ,carbon 14 , Phosphorus 32 , iodine 123 , iodine 125 , iodine 126 , iodine 131 , iodine 133 ,bromine 77 ,technetium 99m ,indium 111,indium 113m ,gallium 67 ,gallium 68 ,ruthenium 95 ,ruthenium 97 ,ruthenium 103 ,ruthenium 105 ,mercury 207 ,mercury 203 ,rhenium 99m ,rhenium 101 ,rhenium 105 ,scandium 47 ,tellurium 121m ,tellurium 122m ,tellurium 125m ,thulium 165 ,thulium 167 ,thulium 168 ,copper 67 , fluorine 18 ,yttrium 90 ,palladium 100 , bismuth 217 and antimony 211 Examples of luminescent labels include radioisotopes such as chemiluminescent (e.g., acridinium ester, luminol, isoluminol) and bioluminescent labels. Examples of immunodetectable labels include haptens, peptides / polypeptides, antibodies, receptors and ligands, such as biotin, avidin, streptavidin, or digoxigenin. Examples of nucleic acid labels include aptamers.

[0246] In some embodiments, the antigen-binding molecule / polypeptide comprises an epitope tag, such as His, (e.g., 6XHis), FLAG, c-Myc, StrepTag, hemagglutinin, E, calmodulin-binding protein (CBP), glutathione-s-transferase (GST), maltose-binding protein (MBP), thioredoxin, S-peptide, T7 peptide, SH2 domain, avidin, streptavidin, and a hapten (e.g., biotin, digoxigenin, dinitrophenol), optionally at the N-terminus or C-terminus of the antigen-binding molecule / polypeptide.

[0247] In some embodiments, the antigen-binding molecule / polypeptide comprises a moiety having a detectable activity, such as an enzymatic moiety, including, for example, luciferase, glucose oxidase, galactosidase (e.g., beta-galactosidase), glucuronidase, phosphatase (e.g., alkaline phosphatase), peroxidase (e.g., horseradish peroxidase), and cholinesterase.

[0248] In some embodiments, an antigen-binding molecule or polypeptide of the present disclosure comprises a chemical moiety. In some embodiments, an antigen-binding molecule / polypeptide of the present disclosure is conjugated to a chemical moiety.

[0249] The chemical moiety can be a moiety for providing a therapeutic effect, i.e., a drug moiety. The drug moiety can be a small molecule (e.g., a low molecular weight (<1000 daltons, typically between about 300 and 700 daltons) organic compound). Drug moieties are described, for example, in Parslow et al., Biomedicines. 2016 Sep;4(3):14, incorporated herein by reference in its entirety. In some embodiments, the drug moiety can be or include a cytotoxic agent. In some embodiments, the drug moiety can be or include a chemotherapeutic agent. Drug moieties include, for example, calicheamicin, DM1, DM4, monomethyl auristatin E (MMAE), monomethyl auristatin F (MMAF), SN-38, doxorubicin, duocarmycin, D6.5, and PBD.

[0250] Nucleic acids and vectors The present disclosure provides one or more nucleic acids encoding an antigen-binding molecule or polypeptide according to the present disclosure. In some embodiments, the nucleic acid(s) comprise or consist of DNA and / or RNA.

[0251] Antigen-binding molecules or polypeptides according to the present disclosure can be produced intracellularly by translation of RNA encoding the polypeptide(s). Antigen-binding molecules or polypeptides according to the present disclosure can be produced intracellularly by transcription from a nucleic acid encoding the polypeptide(s) and subsequent translation of the transcribed RNA.

[0252] In some embodiments, the nucleic acid(s) may be or may be included / contained in one or more vectors. As used herein, a "vector" is a nucleic acid molecule used as a vehicle to transfer exogenous nucleic acid into a cell.

[0253] Therefore, the present disclosure also provides one or more vectors comprising one or more nucleic acids according to the present disclosure. The vector can facilitate the delivery of nucleic acid(s) encoding a polypeptide according to the present disclosure to a cell. The vector may be an expression vector containing elements necessary for expressing a polypeptide according to the present disclosure. The vector may contain elements that facilitate the integration of nucleic acid(s) into the genomic DNA of a cell into which the vector is introduced.

[0254] The nucleic acids and vectors according to the present disclosure can be provided in purified or isolated form, i.e., purified or isolated from other nucleic acids or naturally occurring biological materials.

[0255] The vector may be a vector for expressing a nucleic acid in a cell (i.e., an expression vector). Such a vector may comprise a promoter sequence operably linked to a nucleotide sequence encoding an antigen-binding molecule or polypeptide according to the present disclosure. The vector may also comprise a stop codon (i.e., 3' in the nucleotide sequence of the vector to the nucleotide sequence encoding the polypeptide(s)) and an expression enhancer. Any suitable vector, promoter, enhancer, and stop codon known in the art may be used to express a peptide or polypeptide from a vector according to the present disclosure.

[0256] The term "operably linked" can include a situation in which a nucleic acid encoding a polypeptide according to the present disclosure and a regulatory nucleic acid sequence(s) (e.g., a promoter and / or enhancer) are covalently linked in such a way as to place expression of the nucleic acid encoding the polypeptide under the influence or control of the regulatory nucleic acid sequence(s) (thereby forming an expression cassette). Thus, a regulatory sequence is operably linked to a selected nucleic acid sequence if the regulatory sequence is capable of affecting the transcription of the nucleic acid sequence. The resulting transcript(s) can then be translated into the desired polypeptide(s).

[0257] Vectors contemplated in connection with the present disclosure include DNA vectors, RNA vectors, plasmids (e.g., conjugative plasmids (e.g., F plasmids), non-conjugative plasmids, R plasmids, col plasmids, episomes), viral vectors (e.g., retroviral vectors, e.g., gamma retroviral vectors (e.g., murine leukemia virus (MLV)-derived vectors, e.g., SFG vectors), lentiviral vectors, adenoviral vectors, adeno-associated viral vectors, vaccinia virus vectors, and herpes virus vectors), transposon-based vectors, and artificial chromosomes (e.g., yeast artificial chromosomes), as described, for example, in Maus et al., Annu Rev Immunol (2014) 32:189-225 and Morgan and Boyerinas, Biomedicines (2016) 4:9, both of which are incorporated herein by reference in their entireties. In some embodiments, a vector according to the present disclosure is a lentiviral vector.

[0258] In some embodiments, the vector may be a eukaryotic vector, i.e., a vector that contains elements necessary for protein expression from the vector in eukaryotic cells, hi some embodiments, the vector may be a mammalian vector, for example, one that contains a cytomegalovirus (CMV) or SV40 promoter to drive protein expression.

[0259] The constituent polypeptides of an antigen-binding molecule according to the present disclosure may be encoded by different nucleic acids of a plurality of nucleic acids or by different vectors of a plurality of vectors. Production of antigen-binding molecules and polypeptides Antigen-binding molecules and polypeptides according to the present disclosure can be prepared according to methods for the production of polypeptides known to those skilled in the art.

[0260] Antigen-binding molecules and polypeptides can be prepared by chemical synthesis, for example, liquid phase or solid phase synthesis. For example, peptides / polypeptides can be synthesized using the methods described in, for example, Chandrudu et al., Molecules (2013), 18:4373-4388 (the entire contents of which are incorporated herein by reference).

[0261] Alternatively, antigen-binding molecules and polypeptides can be produced by recombinant expression. Suitable molecular biology techniques for the recombinant production of polypeptides are well known in the art, such as those described in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th Edition), Cold Spring Harbor Press, 2012 and Nat Methods. (2008); 5 (2): 135-146 (both of which are incorporated herein by reference in their entirety). Methods for the recombinant production of antigen-binding molecules are also described in Frenzel et al., Front Immunol. (2013); 4: 217 and Kunert and Reinhart, Appl Microbiol Biotechnol. (2016) 100: 3451-3461, both of which are incorporated herein by reference in their entirety.

[0262] In some cases, the antigen-binding molecules of the present disclosure are composed of more than one polypeptide chain. In such cases, production of the antigen-binding molecule may involve transcription and translation of more than one polypeptide, and subsequent assembly of the polypeptide chains to form the antigen-binding molecule.

[0263] For recombinant production according to the present disclosure, any cell suitable for expression of a polypeptide can be used. The cell can be a prokaryotic or eukaryotic cell. In some embodiments, the cell is a prokaryotic cell, such as an archaeal or bacterial cell. In some embodiments, the bacterium can be a gram-negative bacterium, such as an Enterobacteriaceae bacterium, e.g., Escherichia coli. In some embodiments, the cell is a eukaryotic cell, such as a yeast cell, a plant cell, an insect cell, or a mammalian cell, e.g., a cell described herein above.

[0264] In some cases, the cell is not a prokaryotic cell, because some prokaryotic cells cannot perform the same folding or post-translational modification as eukaryotic cells.In addition, eukaryotic cells can achieve very high expression levels, and proteins can be more easily purified from eukaryotic cells using appropriate tags.Specific plasmids that enhance the secretion of proteins into the medium can also be used.

[0265] In some embodiments, polypeptides can be prepared by cell-free protein synthesis (CFPS), for example, by the system described in Zemella et al., Chembiochem (2015) 16(17):2420-2431, which is incorporated herein by reference in its entirety.

[0266] Production may also involve the culture or fermentation of eukaryotic cells modified to express the polypeptide(s) of interest. The culture or fermentation may be carried out in a bioreactor provided with an appropriate supply of nutrients, air / oxygen, and / or growth factors. Secreted proteins may be recovered by separating the culture medium / fermentation broth from the cells, extracting the protein content, and separating the individual proteins to isolate the secreted polypeptide(s). Culture, fermentation, and separation techniques are well known to those skilled in the art and are described, for example, in Green and Sambrook, Molecular Cloning: A Laboratory Manual (4th ed.; incorporated herein by reference).

[0267] A bioreactor contains one or more containers in which cells can be cultured. Cultivation in a bioreactor can be carried out continuously, with a continuous inflow of reactants into the reactor and a continuous outflow of cultured cells from the reactor. Alternatively, cultivation can be carried out in batches. Bioreactors monitor and regulate environmental conditions within the container, such as pH, oxygen, inflow and outflow rates, and agitation, to provide optimal conditions for the cultured cells.

[0268] Following culturing of cells expressing the polypeptide(s), the polypeptide(s) of interest can be isolated. Any suitable method known in the art for separating proteins from cells can be used. To isolate the polypeptide, it may be necessary to separate the cells from the nutrient medium. If the polypeptide(s) are secreted from the cells, the cells can be separated from the culture medium containing the secreted polypeptide(s) of interest by centrifugation. If the polypeptide(s) of interest are concentrated within the cells, protein isolation can include centrifugation to separate the cells from the cell culture medium, treating the cell pellet with a lysis buffer, and disrupting the cells, for example, by sonication, rapid freeze-thawing, or osmotic lysis.

[0269] It may then be desirable to isolate the polypeptide(s) of interest from the supernatant or culture medium, which may contain other proteins and non-protein components. A common technique for separating protein components from the supernatant or culture medium is by precipitation. Proteins of different solubilities are precipitated with different concentrations of a precipitant, such as ammonium sulfate. For example, low concentrations of the precipitant extract soluble proteins. Thus, by adding increasing concentrations of the precipitant, proteins of different solubilities can be distinguished. Dialysis can then be used to remove ammonium sulfate from the separated proteins.

[0270] Other methods for distinguishing different proteins are known in the art, such as ion exchange chromatography and size chromatography, which can be used as alternatives to precipitation or can be performed following precipitation.

[0271] Once the polypeptide(s) of interest have been isolated from the culture, it may be desirable or necessary to concentrate the polypeptide(s). Numerous methods for concentrating proteins are known in the art, such as ultrafiltration or lyophilization.

[0272] Antigen-binding molecules and cells containing / expressing polypeptides The present disclosure also provides cells containing or expressing an antigen-binding molecule or polypeptide according to the present disclosure, as well as cells containing or expressing one or more nucleic acids, one or more vectors according to the present disclosure.

[0273] It will be understood that where a cell is referred to herein in the singular (i.e., "a / the cell"), a plurality / population of such cells is also contemplated. The cell may be a eukaryotic cell, for example, a mammalian cell. The mammal may be a primate (rhesus monkey, cynomolgus monkey, non-human primate, or human) or a non-human mammal (e.g., rabbit, guinea pig, rat, mouse, or other rodent (including any animal in the order Rodents), cat, dog, pig, sheep, goat, cow (including cows, e.g., dairy cows, or any animal in the order Bos), horse (any animal in the order Equidae), donkey, and non-human primate).

[0274] In some embodiments, the cells are or are derived from a cell type commonly used for the expression of polypeptides for therapeutic use in humans. Exemplary cells are described, for example, in Kunert and Reinhart, Appl Microbiol Biotechnol. (2016) 100:3451-3461 (incorporated herein by reference in its entirety), and include, for example, CHO, HEK293, PER.C6, NS0, and BHK cells. In a preferred embodiment, the cells are or are derived from CHO cells.

[0275] The present disclosure also provides methods for producing a cell comprising a nucleic acid(s) or vector(s) according to the present disclosure, the method comprising introducing one or more nucleic acids, one or more vectors according to the present disclosure into a cell. In some embodiments, introducing the isolated nucleic acid(s) or vector(s) according to the present disclosure into a cell comprises transformation, transfection, electroporation, or transduction (e.g., retroviral transduction).

[0276] The present disclosure also provides a method for producing a cell that expresses / contains an antigen-binding molecule or polypeptide according to the present disclosure, the method comprising introducing one or more nucleic acids, one or more vectors according to the present disclosure into the cell. In some embodiments, the method further comprises culturing the cell under conditions suitable for expression of the nucleic acid(s) or vector(s) by the cell. In some embodiments, the method is performed in vitro.

[0277] The present disclosure also provides cells obtained or obtainable by a method according to the present disclosure. Combinations containing known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecules The present disclosure also provides a combination comprising (i) an antigen-binding molecule according to the present disclosure and (ii) a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule. The present disclosure also provides a composition comprising (i) an antigen-binding molecule according to the present disclosure and (ii) a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule.

[0278] It will be understood that the antigen-binding molecule of (i) according to the preceding paragraph may be an antigen-binding molecule according to any embodiment described in the section of the present specification entitled "Antigen-binding molecules of the present disclosure." It will likewise be understood that the antigen-binding molecule of (ii) according to the preceding paragraph may be a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule according to any embodiment described in the section of the present specification entitled "Known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecules."

[0279] In some embodiments, the combinations / compositions of the present disclosure comprise (i) an antigen binding molecule according to one of (1) to (19) in the section entitled "Detailed Exemplary Antigen Binding Molecules" and (ii) an antigen binding molecule according to one of (A) to (D) in the section entitled "Known SARS-CoV-2 Spike Protein / SARS-CoV-2 Variant Spike Protein-Binding Antigen Binding Molecules."

[0280] In some aspects and embodiments, the combination is a pharmaceutical combination.As used herein, "pharmaceutical combination" refers to a product that contains multiple (typically two herein) different active (i.e., therapeutic / prophylactic) agents that are intended to be used in combination.The drugs of pharmaceutical combination can be formulated together or separately, but are typically packaged with a package insert that includes instructions for the use of the drugs in combination.

[0281] In some embodiments, the agents of the pharmaceutical combination are contained in a single composition, for example, a pharmaceutical composition containing both agents. In some embodiments, the agents of the pharmaceutical combination are contained in separate compositions; for example, a pharmaceutical combination may be provided as (i) a pharmaceutical composition containing an antigen-binding molecule according to the present disclosure, and (ii) a pharmaceutical composition containing a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule.

[0282] The present disclosure also provides compositions (e.g., pharmaceutical compositions and medicaments) comprising the agents described herein (i.e., (i) and (ii) above). Such compositions may include related articles in formulations suitable for clinical use.

[0283] The present disclosure also provides combinations (and compositions comprising the combinations) of antigen-binding molecules according to (A) or (B) with antigen-binding molecules according to (C) or (D) (i.e., as described in the section herein entitled "Known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecules"). In some embodiments, the combinations comprise antigen-binding molecules according to (A) and antigen-binding molecules according to (B). In some embodiments, the combinations comprise antigen-binding molecules according to (C) and antigen-binding molecules according to (D).

[0284] Functional Properties of the Combinations of the Present Disclosure The combinations described herein and compositions comprising such combinations (hereinafter in this section "combination(s) / composition(s)") can be characterized with reference to specific functional properties. In some embodiments, the combinations described herein exhibit the following properties: inhibiting the interaction between a sarbecovirus spike protein (e.g., SARSr-CoV spike protein; e.g., SARS-CoV-2 spike protein and / or one or more SARS-CoV-2 variant spike proteins) and ACE2 with increased potency (e.g., a lower IC50) compared to the potency with which such interaction is inhibited by the constituent agents of the combination / composition when used alone; and / or inhibiting infection of ACE2-expressing cells by a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 and / or one or more SARS-CoV-2 variants) with increased potency (e.g., a lower IC50) compared to the potency with which such infection is inhibited by the components of the combination / composition when used alone; may have one or more of:

[0285] In some embodiments, the combinations / compositions of the disclosure measure the interaction between SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein and ACE2 by measuring the IC for inhibition of the interaction between the relevant protein and ACE2 by the constituent agents of the combination / composition when used alone, as determined in the same assay. 50 an IC that is less than 1 times the original concentration, for example, ≦0.99 times, ≦0.95 times, ≦0.9 times, ≦0.85 times, ≦0.8 times, ≦0.75 times, ≦0.7 times, ≦0.65 times, ≦0.6 times, ≦0.55 times, ≦0.5 times, ≦0.45 times, ≦0.4 times, ≦0.35 times, ≦0.3 times, ≦0.25 times, ≦0.2 times, ≦0.15 times, ≦0.1 times, ≦0.05 times, or ≦0.01 times the original concentration 50 and inhibits.

[0286] In some embodiments, the combinations / compositions of the present disclosure inhibit infection of ACE2-expressing cells by a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 and / or one or more SARS-CoV-2 variants) by the IC50 or IC60 inhibitors of the combination / composition, when used alone, for inhibition of infection of such cells by the relevant SARSr-CoV, as determined in the same assay. 50 an IC that is less than 1 times the original concentration, for example, ≦0.99 times, ≦0.95 times, ≦0.9 times, ≦0.85 times, ≦0.8 times, ≦0.75 times, ≦0.7 times, ≦0.65 times, ≦0.6 times, ≦0.55 times, ≦0.5 times, ≦0.45 times, ≦0.4 times, ≦0.35 times, ≦0.3 times, ≦0.25 times, ≦0.2 times, ≦0.15 times, ≦0.1 times, ≦0.05 times, or ≦0.01 times the original concentration 50 and inhibits.

[0287] In some embodiments, the combinations / compositions according to the present disclosure achieve synergistic inhibition of the interaction between the SARS-CoV-2 spike protein and / or a given SARS-CoV-2 variant spike protein and ACE2. In some embodiments, the combinations / compositions according to the present disclosure achieve synergistic inhibition of infection of ACE2-expressing cells by a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 and / or one or more SARS-CoV-2 variants). That is, in some embodiments, the combinations / compositions achieve a level of inhibition that is synergistic (i.e., greater than additive) compared to that observed when an antigen binding molecule of the present disclosure is used alone and / or compared to that observed when a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen binding molecule is used alone.

[0288] As used herein, a "synergistic" or "greater than additive" level of relative effect (e.g., inhibition of interaction, inhibition of infection) for a given combination / composition refers to a level of effect that is greater than the sum of the effects observed when the individual components of the combination / composition are used alone.

[0289] Quantitative methods for assessing synergy are described, for example, in Tallarida, Genes Cancer. (2011) 2(11):1003-1008 and Chou, Cancer Res (2010) 70:440-446, both of which are incorporated herein by reference in their entireties. Additive, synergistic, and antagonistic effects can be evaluated in experiments in which various dose ranges of the combination / composition and its individual components are evaluated for the associated effect. Dose-response curves can be plotted and evaluated to determine whether the combination / composition achieves a synergistic level of associated effect compared to the individual components of the combination / composition used alone. In some embodiments, synergistic effects can be assessed using combination / composition index (CI) values ​​calculated using the Chou-Talalay method described in Chou, Cancer Res (2010) 70:440-446. According to the Chow-Talalay method, a CI=1 for a given combination / composition indicates an additive effect, a CI<1 indicates a synergistic effect, and a CI>1 indicates antagonism.

[0290] composition The present disclosure also provides compositions comprising the antigen-binding molecules, polypeptides, nucleic acids, expression vectors and / or cells described herein.

[0291] The antigen-binding molecules, polypeptides, nucleic acids, expression vectors, and cells described herein may be formulated as pharmaceutical compositions or medicaments for clinical use and may contain pharmaceutically acceptable carriers, diluents, excipients, or adjuvants. Thus, the present disclosure also provides pharmaceutical compositions / medicines comprising the antigen-binding molecules, polypeptides, nucleic acid(s), expression vector(s), or cells described herein.

[0292] The compositions of the present disclosure may be formulated in any combination with one or more pharmaceutically acceptable carriers (e.g., liposomes, micelles, microspheres, nanoparticles), diluents / excipients (e.g., starch, cellulose, cellulose derivatives, polyols, dextrose, maltodextrin, magnesium stearate), adjuvants, fillers, buffering agents, preservatives (e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium, cysteine, methionine, citric acid, sodium citrate, methylparaben, propylparaben), anti-inflammatory agents, or the like. They may contain oxidizing agents (e.g., vitamin A, vitamin E, vitamin C, retinyl palmitate, selenium), lubricants (e.g., magnesium stearate, talc, silica, stearic acid, vegetable stearin), binders (e.g., sucrose, lactose, starch, cellulose, gelatin, polyethylene glycol (PEG), polyvinylpyrrolidone (PVP), xylitol, sorbitol, mannitol), stabilizers, solubilizers, surfactants (e.g., wetting agents), masking agents, or colorants (e.g., titanium dioxide).

[0293] As used herein, the term "pharmaceutically acceptable" refers to compounds, ingredients, materials, compositions, dosage forms, etc. that are suitable for use in contact with the tissues of an applicable subject (e.g., a human subject) without undue toxicity, irritation, allergic response, or other problem or complication, and with a reasonable benefit / risk ratio, within the scope of sound medical judgment. Each carrier, diluent, excipient, adjuvant, filler, buffer, preservative, antioxidant, lubricant, binder, stabilizer, solubilizer, surfactant, masking agent, colorant, flavoring, or sweetener of a composition according to the present disclosure must also be "acceptable" in the sense of being compatible with the other ingredients of the formulation. Suitable carriers, diluents, excipients, adjuvants, fillers, buffers, preservatives, antioxidants, lubricants, binders, stabilizers, solubilizers, surfactants, masking agents, colorants, flavorings, or sweetening agents can be found in standard pharmaceutical texts, such as Remington's 'The Science and Practice of Pharmacy' (Ed. A. Adejare), 23rd Edition (2020), Academic Press.

[0294] The composition may be formulated for topical, parenteral, systemic, intracavity, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intraconjunctival, subcutaneous, intradermal, intrathecal, oral or transdermal administration. In some embodiments, the pharmaceutical composition / medicament may be formulated for administration by injection or infusion, or by oral ingestion.

[0295] Suitable formulations may contain the relevant items in a sterile or isotonic medium.Medicinal and pharmaceutical compositions may be formulated into liquid forms, including gels.Liquid formulations may be formulated for administration by injection or infusion (e.g., using a catheter) into selected areas of the human or animal body.

[0296] In some embodiments, the composition is formulated, for example, for injection or infusion into a blood vessel, tissue / organ of interest. The present disclosure also provides methods for producing pharmaceutically useful compositions and medicaments. Such methods may include one or more steps selected from the following: producing the antigen-binding molecules, polypeptides, nucleic acids(or nucleic acids), expression vectors(or vectors), or cells described herein; isolating the antigen-binding molecules, polypeptides, nucleic acids(or nucleic acids), expression vectors(or vectors), or cells described herein; and / or mixing the antigen-binding molecules, polypeptides, nucleic acids(or nucleic acids), expression vectors(or vectors), or cells described herein with a pharmaceutically acceptable carrier, adjuvant, excipient, or diluent.

[0297] For example, a further aspect of the present disclosure relates to a method of formulating or producing a medicament or pharmaceutical composition for use in treating a disease / condition (e.g., a disease / condition described herein), the method comprising formulating the pharmaceutical composition or medicament by mixing an antigen-binding molecule, polypeptide, nucleic acid(s), expression vector(s), or cell(s) described herein with a pharmaceutically acceptable carrier, adjuvant, excipient, or diluent.

[0298] Therapeutic and preventive applications The antigen-binding molecules, polypeptides, nucleic acids, expression vectors, cells, combinations and compositions described herein are used in therapeutic and prophylactic methods.

[0299] The present disclosure provides antigen-binding molecules, polypeptides, nucleic acids (or nucleic acids), expression vectors (or vectors), cells, compositions, or combinations described herein for use in methods of medical treatment or prevention. Similarly, antigen-binding molecules, polypeptides, nucleic acids (or nucleic acids), expression vectors (or vectors), cells, compositions, or combinations described herein for use in methods of treating or preventing the diseases or conditions described herein are provided. Also provided is the use of antigen-binding molecules, polypeptides, nucleic acids (or nucleic acids), expression vectors (or vectors), cells, compositions, or combinations described herein in the manufacture of a medicament for treating or preventing the diseases or conditions described herein. Also provided is a method of treating or preventing the diseases or conditions described herein, comprising administering to a subject a therapeutically or prophylactically effective amount of an antigen-binding molecule, polypeptide, nucleic acid (or nucleic acids), expression vectors (or vectors), cells, compositions, or combinations described herein.

[0300] The present disclosure also provides an antigen-binding molecule according to the present disclosure for use in a method for treating or preventing a disease / condition described herein, wherein the method further comprises administering a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule. Similarly, the present disclosure also provides a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule for use in a method for treating or preventing a disease / condition described herein, wherein the method further comprises administering an antigen-binding molecule according to the present disclosure.

[0301] Also provided is the use of an antigen-binding molecule according to the present disclosure in the manufacture of a medicament for use in a method of treating or preventing a disease / condition described herein, wherein the method further comprises administering a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule.Also provided is the use of a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule in the manufacture of a medicament for use in a method of treating or preventing a disease / condition described herein, wherein the method further comprises administering an antigen-binding molecule according to the present disclosure.

[0302] Further provided are methods for treating or preventing the diseases / conditions described herein, comprising administering to a subject in need thereof a therapeutically or prophylactically effective amount of (i) an antigen-binding molecule according to the present disclosure and (ii) a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule.

[0303] The present disclosure also provides (i) an antigen-binding molecule according to the present disclosure and (ii) a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule for use in a method of treating or preventing a disease / condition described herein in a subject. Similarly, the present disclosure also provides use of (i) an antigen-binding molecule according to the present disclosure and (ii) a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule in the manufacture of a medicament for use in treating or preventing a disease / condition described herein in a subject. Also provided is a method of treating or preventing a disease / condition described herein in a subject, comprising administering to the subject a therapeutically or prophylactically effective amount of (i) an antigen-binding molecule according to the present disclosure and (ii) a known SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein-binding antigen-binding molecule.

[0304] In embodiments according to the aspects of the preceding paragraph, the provision of (i) and (ii) may be as a combination therapy. In some embodiments, (i) and (ii) may be provided simultaneously or sequentially.

[0305] The articles, methods, and uses of the present disclosure may be effective for reducing the development or progression of a disease / condition, alleviating the symptoms of a disease / condition, or reducing the pathology of a disease / condition. The articles, methods, and uses may be effective for preventing the progression of a disease / condition, for example, preventing the worsening of a disease / condition or slowing its rate of development. In some embodiments, the articles, methods, and uses may result in an improvement in a disease / condition, for example, a reduction in the symptoms of a disease / condition or some other related reduction in the severity / activity of a disease / condition. In some embodiments, the articles, methods, and uses may prevent the disease / condition from progressing to a later stage (e.g., a chronic stage).

[0306] It will be understood that the articles of the present disclosure can be used for the treatment / prevention of any disease / condition that would derive a therapeutic or prophylactic benefit from a reduction in the levels of a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) or a reduction in the number of cells infected with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant). For example, the disease / condition can be one in which infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) is pathologically associated, e.g., infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) is positively associated with the onset, development, or progression of the disease / condition and / or the severity of one or more symptoms of the disease / condition, or infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) is a risk factor for the onset, development, or progression of the disease / condition.

[0307] In some embodiments, the disease / condition treated / prevented in accordance with the present disclosure is a disease / condition characterized by infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant), e.g., COVID-19. In some embodiments, the disease / condition is a disease / condition caused by infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant), e.g., COVID-19.

[0308] Clinical characteristics of COVID-19 are described in Lechien et al., Journal of Internal Medicine (2020) 288(3):335-344; International Severe Acute Respiratory and Emerging Infections Consortium (ISARIC). COVID-19 Report: 19 May 2020: ISARIC; 2020; Docherty et al., BMJ (2020) 369:m1985; and Bhardwaj et al., Int Rev Immunol. (2021) 2021:1-36, all of which are incorporated herein by reference in their entireties. Common symptoms include cough, fever, headache, dyspnea, anosmia, pharyngitis, nasal congestion, rhinorrhea, asthenia, myalgia, arthralgia, taste dysfunction, abdominal pain, vomiting, and diarrhea. Most patients present with mild or moderate illness, but hospitalization occasionally is required, especially in elderly patients and / or those with comorbid conditions such as diabetes and cardiovascular disease. The main complication of COVID-19 is the progression to acute respiratory distress syndrome (ARDS), which manifests as respiratory distress and acute respiratory failure requiring mechanical ventilation in patients. Some infected patients are asymptomatic.

[0309] Treatment according to the methods of the disclosure can result in a reduction in the level or viral load of a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) in the subject or in the tissue / organ (e.g., lung) of the subject, a reduction in the expression level of a pro-inflammatory factor (e.g., IL-6, CCL2 and / or CXCL10) in the subject or in the tissue / organ (e.g., lung) of the subject, an increase in the level of expression of IFNγ in the subject or in the tissue / organ (e.g., lung) of the subject, an increase in the level of expression of a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) in the subject or in the tissue / organ (e.g., lung) of the subject, an increase in the level of expression of a pro-inflammatory factor (e.g., IL-6, CCL2 and / or CXCL10) ... pro-inflammatory factor (e.g., IL-6, CCL2 and / or CXCL10) in the subject or in the tissue / organ (e.g., lung) of the subject, an increase in the level of expression of a pro-inflammatory factor (e.g., IL-6, CCL In one or more of the methods, the present invention can achieve one or more of the following: a reduction in the number / percentage of cells infected with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant), inhibition of the development / progression of a disease / condition caused by infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant), e.g., COVID-19, in a subject; a reduction in the severity of symptoms of a disease / condition caused by infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant), e.g., COVID-19, in a subject; inhibition of the development / progression of acute respiratory distress syndrome (ARDS) in a subject; and an increase in survival rate of the subject.

[0310] In some embodiments, a subject may be selected for a treatment described herein based on a determination of infection with a sarbecovirus (e.g., SARSr-CoV, e.g., a SARS-CoV-2 / SARS-CoV-2 variant), e.g., by detection of a sarbecovirus (e.g., a SARSr-CoV, e.g., a SARS-CoV-2 / SARS-CoV-2 variant) in a sample obtained from the subject. In some embodiments, a subject may be selected for a treatment described herein based on a determination that the subject is at risk for infection with a sarbecovirus (e.g., a SARSr-CoV, e.g., a SARS-CoV-2 / SARS-CoV-2 variant). For example, the subject may have been in close contact with a subject infected with a sarbecovirus (e.g., a SARSr-CoV, e.g., a SARS-CoV-2 / SARS-CoV-2 variant).

[0311] Administration of the articles of the present disclosure is preferably in a "therapeutically effective" or "prophylactically effective" amount, which is sufficient to provide a therapeutic or prophylactic benefit to the subject. The actual amount administered and the rate and time course of administration will depend on the nature and severity of the disease / condition and the specific article being administered. Prescribing treatment, e.g., determining dosage, is within the responsibility of the practitioner or other physician and typically takes into account the disease / disorder being treated, the condition of the individual subject, the site of delivery, the method of administration, and other factors known to the practitioner. Examples of the techniques and protocols mentioned above can be found in Remington's 'The Science and Practice of Pharmacy' (ed. A. Adejare), 23rd Edition (2020), Academic Press.

[0312] Administration of the articles of the present disclosure may be parenteral, systemic, intracavitary, intravenous, intraarterial, intramuscular, intrathecal, intraocular, intravitreal, intraconjunctival, subretinal, suprachoroidal, subcutaneous, intradermal, intrathecal, oral, nasal, topical, or transdermal. Administration may be by any suitable mode of nasal delivery, such as nasal drops, nasal spray, nebulizer, etc. Administration may be by injection or infusion.

[0313] Multiple doses of the antigen-binding molecules, polypeptides, nucleic acids (or nucleic acids), expression vectors (or nucleic acids), cells, compositions, or combinations described herein may be provided. The multiple doses may be separated by a predetermined time interval, which may be selected to be 1, 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, or 31 days, or 1, 2, 3, 4, 5, or 6 months. By way of example, a dose may be given once every 7, 14, 21, or 28 days (plus or minus 3, 2, or 1 day).

[0314] Administration of the articles of the present disclosure may be either alone or in combination with additional prophylactic / therapeutic agents, either simultaneously or sequentially, depending on the disease / condition being treated. The antigen-binding molecules, cells, compositions or combinations described herein and the additional prophylactic / therapeutic agents may be administered simultaneously or sequentially.

[0315] Concurrent administration refers to administration of an antigen-binding molecule, polypeptide, nucleic acid(s), expression vector(s), cell(s), composition, or combination of the present disclosure and an additional prophylactic / therapeutic agent together, for example, as a pharmaceutical composition (combined preparation) containing both agents, or immediately following each other, optionally via the same administration route, for example, into the same artery, vein, or other blood vessel. Sequential administration refers to administration of either (i) an antigen-binding molecule, polypeptide, nucleic acid(s), expression vector(s), cell(s), composition, or combination of the present disclosure, or (ii) an additional prophylactic / therapeutic agent, followed by the separate administration of the other (i) / (ii) after a given time interval. Although this is the case in some embodiments, (i) and (ii) need not be administered by the same route. The time interval can be any time interval.

[0316] The present disclosure further provides uses of antigen-binding molecules / combinations / compositions according to the present disclosure for inhibiting the interaction between a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) and ACE2; and / or for inhibiting infection of ACE2-expressing cells by SARS-CoV-2 / SARS-CoV-2 variants. The present disclosure further provides methods for inhibiting the interaction between a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) and ACE2 and / or for inhibiting infection of ACE2-expressing cells by SARS-CoV-2 / SARS-CoV-2 variants using antigen-binding molecules / combinations / compositions according to the present disclosure. Such uses / methods may be in vitro or in vivo in a subject.

[0317] Accordingly, the present disclosure provides a method for inhibiting the interaction between a sarbecovirus spike protein (e.g., a SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) and ACE2 and / or inhibiting infection of ACE2-expressing cells by SARS-CoV-2 / SARS-CoV-2 variants, comprising administering to a subject an antigen-binding molecule / combination / composition according to the present disclosure.

[0318] Detection Method The present disclosure also provides articles of the present disclosure for use in methods for detecting, identifying, or imaging sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein), or cells comprising sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein), e.g., as a result of infection with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant).

[0319] The antigen-binding molecules, combinations, and compositions described herein can be used in methods comprising detecting binding of the antigen-binding molecule to a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein). Such methods can include detecting binding complexes between the antigen-binding molecule and a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein). It will be understood that the sarbecovirus / sarbecovirus spike protein can be contained in a cell, for example, as a result of infection of the cell by the sarbecovirus.

[0320] Thus, provided is a method comprising contacting a sample containing or suspected of containing a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) with an antigen binding molecule / combination / composition according to the present disclosure, and detecting formation of a complex between the antigen binding molecule and the sarbecovirus / sarbecovirus spike protein. Similarly, methods are provided that include contacting a sample containing or suspected of containing a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or cells comprising a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) with an antigen-binding molecule / combination / composition according to the present disclosure, and detecting formation of a complex between the antigen-binding molecule and the sarbecovirus / sarbecovirus spike protein.

[0321] Suitable method formats are well known in the art and include immunoassays such as sandwich assays, for example, ELISA. The method may include labeling the antigen-binding molecule or target(s), or both, with a detectable moiety, such as a fluorescent label, phosphorescent label, luminescent label, immunodetectable label, radiolabel, chemical, nucleic acid, or enzymatic label described herein. Detection techniques are well known to those skilled in the art and can be selected depending on the labeling agent.

[0322] Methods comprising detecting a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein), or a cell comprising a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein), include methods for diagnosing / prognosing the diseases / conditions described herein.

[0323] Such methods may be performed in vitro on a patient sample or after processing of the patient sample. Once the sample is collected, the patient need not be present for the in vitro method to be performed, and therefore the method may not be performed in the human or animal body. In some embodiments, the method is performed in vivo.

[0324] Such methods can include, for example, detecting or quantifying sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) and / or cells containing sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein) in a patient sample. When the method includes quantifying a relevant factor, the method can further include comparing the determined amount to a standard or reference value as part of a diagnostic or prognostic evaluation. Other diagnostic / prognostic tests can be used in conjunction with those described herein to enhance the accuracy of the diagnosis or prognosis or to confirm the results obtained by using the tests described herein.

[0325] Detection in a sample can be used for purposes of diagnosis of a disease / condition (e.g., COVID-19), a predisposition to a disease / condition, or to provide a prognosis (prognosis) for a disease / condition, such as a disease / condition described herein. The diagnosis or prognosis may relate to an existing (already diagnosed) disease / condition.

[0326] The sample can be collected from any tissue or body fluid. The sample obtained from the subject can be of any type. The biological sample can be collected from any tissue or body fluid, such as a blood sample, a blood-derived sample, a serum sample, a lymph sample, a semen sample, a saliva sample, or a synovial fluid sample. The blood-derived sample can be a selected fraction of the patient's blood, such as a fraction containing selected cells, or a plasma or serum fraction. The sample can include a tissue sample or a biopsy; or cells isolated from the subject.

[0327] Subjects may be selected for diagnostic / prognostic evaluation based on the presence of symptoms indicative of a disease / condition described herein, or based on the subject being considered to be at risk for developing a disease / condition described herein.

[0328] The present disclosure also provides methods for selecting / stratifying subjects for treatment with a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) targeting agent. In some embodiments, a subject is selected for treatment / prevention by a method of the present disclosure, or identified as a subject who would benefit from such treatment / prevention, based on detection / quantification, e.g., in a sample obtained from the individual, of a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein), or cells containing a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein).

[0329] subject The subject according to various aspects of the present disclosure can be any animal or human. Therapeutic or prophylactic applications can be in humans or animals (veterinary use).

[0330] A subject to which an article of the present disclosure is administered (e.g., via therapeutic or prophylactic intervention) may be a subject in need of such intervention. The subject is preferably a mammal, more preferably a human. The subject may be a non-human mammal, but is more preferably a human. The subject may be male or female. The subject may be a patient.

[0331] A subject may have (e.g., may have been diagnosed with), may be suspected of having, or may be at risk of developing / suffering from a disease or condition described herein. In embodiments according to the present disclosure, a subject may be selected for treatment by a method based on characterization for one or more markers for such a disease / condition.

[0332] In some embodiments, a subject may be selected for a therapeutic or preventative intervention described herein based on detection of a sarbecovirus (e.g., SARSr-CoV, e.g., SARS-CoV-2 / SARS-CoV-2 variant) and / or a sarbecovirus spike protein (e.g., SARS-CoV-2 spike protein / SARS-CoV-2 variant spike protein), e.g., in a sample obtained from the subject.

[0333] kit The present disclosure also provides a kit of parts. In some embodiments, the kit may have at least one container with a predetermined amount of an antigen-binding molecule, polypeptide, nucleic acid(s), expression vector(s), cell, composition, or combination described herein.

[0334] In some embodiments, the kit may include materials for producing an antigen-binding molecule, polypeptide, nucleic acid(s), expression vector(s), cell, composition, or combination described herein. In some embodiments, the kit of parts may include materials for formulating an antigen-binding molecule, polypeptide, nucleic acid(s), expression vector(s), cell, composition, or combination described herein into a pharmaceutical composition / medicament, e.g., in a composition further comprising a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.

[0335] The kit can be provided with instructions for administering the antigen-binding molecule, polypeptide, nucleic acid(s), expression vector(s), cell, composition or combination to a patient to treat a particular disease / condition (e.g., a disease / condition described herein).

[0336] In some embodiments, the kit may further comprise at least one container containing a predetermined amount of another therapeutic agent (e.g., as described herein.) In such embodiments, the kit may also include a second medicament or pharmaceutical composition, such that the two medicaments or pharmaceutical compositions can be administered simultaneously or separately to provide a combined treatment for a particular disease / condition.

[0337] Kits according to the present disclosure may include instructions for use, for example, in the form of an instruction booklet or leaflet. The instructions may include protocols for carrying out any one or more of the methods described herein.

[0338] Sequence identity As used herein, "sequence identity" refers to the percentage of nucleotides / amino acid residues in a subject sequence that are identical to nucleotides / amino acid residues in a reference sequence after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity between the sequences. Pairwise and multiple sequence alignments for determining the percent sequence identity between two or more amino acid or nucleic acid sequences can be accomplished in a variety of ways known to those skilled in the art, for example, using publicly available computer software such as ClustalOmega (Soding, J. 2005, Bioinformatics 21, 951-960), T-coffee (Notredame et al., 2000, J. Mol. Biol. (2000) 302, 205-217), Kalign (Lassmann and Sonnhammer 2005, BMC Bioinformatics, 6 (298)) and MAFFT (Katoh and Standley 2013, Molecular Biology and Evolution, 30 (4) 772-780). When using such software, default parameters are preferably used, for example, for gap penalties and extension penalties.

[0339] array

[0340] [Table 4-1]

[0341] [Table 4-2]

[0342] [Table 4-3]

[0343] [Table 4-4]

[0344] Table 4-5

[0345] Table 4-6

[0346] Table 4-7

[0347] Table 4-8

[0348] Table 4-9

[0349] Table 4-10

[0350] Table 4-11

[0351] Table 4-12

[0352] Table 4-13

[0353] Table 4-14

[0354] Table 4-15

[0355] Table 4-16

[0356] Table 4-17

[0357] Table 4-18

[0358] Table 4-19

[0359] Table 4-20

[0360] Table 4-21

[0361] Table 4-22

[0362] Table 4-23

[0363] Table 4-24

[0364] Table 4-25

[0365] Table 4-26

[0366] Table 4-27

[0367] Table 4-28

[0368] Table 4-29

[0369] Table 4-30

[0370] Table 4-31

[0371] Table 5

[0372] Table 6-1

[0373] Table 6-2

[0374] Table 7

[0375] [Table 8]

[0376] Further aspects and embodiments of the present disclosure The following sections describe further aspects, embodiments and technical features according to the present disclosure. A wide range of protein antigen-binding molecules, such as neutralizing antibodies, suitable for use in the treatment or prevention of coronavirus infection, particularly SARS-CoV-2 variants, are anticipated.

[0377] Accordingly, embodiments of the present disclosure refer to antigen binding molecules that bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB at concentrations less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB, wherein the antigen binding molecule comprises (i) a heavy chain having amino acids with at least 95% sequence identity to SEQ ID NO: 822 or SEQ ID NO: 36 or SEQ ID NO: 52; and (ii) a light chain having amino acids with at least 95% sequence identity to SEQ ID NO: 823 or SEQ ID NO: 44 or SEQ ID NO: 59.

[0378] According to another embodiment, there is an antigen binding molecule that binds to and neutralizes SARS-CoV-2 variant BQ.1.1 at a concentration of E7 that is less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant BQ.1.1; at least one other SARS-CoV-2 variant and another sarbecovirus, wherein the antigen binding molecule is selected from the group consisting of (i) SEQ ID NO:822, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:67, SEQ ID NO:83, SEQ ID NO:96, SEQ ID NO:105, SEQ ID NO:120, SEQ ID NO: 136, SEQ ID NO: 149, SEQ ID NO: 179, SEQ ID NO: 193, SEQ ID NO: 206, SEQ ID NO: 220, SEQ ID NO: 285, SEQ ID NO: 299, SEQ ID NO: 312, SEQ ID NO: 325, SEQ ID NO: 336, SEQ ID NO: 362, SEQ ID NO: 368, SEQ ID NO: 381, SEQ ID NO: 393, SEQ ID NO: 436, SEQ ID NO: 464, SEQ ID NO: 496, SEQ ID NO: 508, SEQ ID NO: 522, SEQ ID NO: 535, SEQ ID NO: 547, SEQ ID NO: 559, SEQ ID NO: 584, SEQ ID NO: 592, SEQ ID NO: 600, SEQ ID NO: 614, and (ii) a light chain having at least 95% sequence identity to an amino acid selected from any one of SEQ ID NO:823, SEQ ID NO:44, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:89, SEQ ID NO:102, SEQ ID NO:113, SEQ ID NO:128, SEQ ID NO:144, SEQ ID NO:157, SEQ ID NO:187, SEQ ID NO:200, SEQ ID NO:213, SEQ ID NO:228, SEQ ID NO:291, SEQ ID NO:305, SEQ ID NO:318, SEQ ID NO:331, SEQ ID NO:343, SEQ ID NO:366, SEQ ID NO:374, SEQ ID NO:388, SEQ ID NO:400, SEQ ID NO:443, SEQ ID NO:471, SEQ ID NO:502, SEQ ID NO:515, SEQ ID NO:529, SEQ ID NO:542, SEQ ID NO:554, SEQ ID NO:567, SEQ ID NO:587, SEQ ID NO:596, SEQ ID NO:607, SEQ ID NO:621, SEQ ID NO:635, SEQ ID NO:651, and SEQ ID NO:663.

[0379] According to another embodiment, there is a composition of any one of the antigen binding molecules described herein above and bebuterovimab LY-CoV1404 and E7. According to another aspect, there is a method of treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of an antigen-binding molecule or composition described herein.

[0380] According to another embodiment, there is a therapeutically effective amount of an antigen binding molecule or composition described herein above for use in treating a sarbecovirus infection. According to various embodiments, there is an antigen binding molecule that binds to and neutralizes SARS-CoV-2 variants BQ.1.1 and XBB at a concentration of E7 that is less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB, wherein the antigen binding molecule comprises (i) a heavy chain having amino acids with at least 95% sequence identity to SEQ ID NO: 822 or SEQ ID NO: 36 or SEQ ID NO: 52 or SEQ ID NO: 96; and (ii) a light chain having amino acids with at least 95% sequence identity to SEQ ID NO: 823 or SEQ ID NO: 44 or SEQ ID NO: 59 or SEQ ID NO: 102.

[0381] In various embodiments, (i) the VH region incorporates the following CDRs: HC-CDR1 having amino acids with at least 85% sequence identity to SEQ ID NO: 37; HC-CDR2 having amino acids with at least 85% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 53; HC-CDR3 having amino acids with at least 85% sequence identity to SEQ ID NO: 39 or SEQ ID NO: 54; and (ii) the VL region incorporates the following CDRs: LC-CDR1 having amino acids with at least 85% sequence identity to SEQ ID NO: 45 or SEQ ID NO: 60; LC-CDR2 having amino acids with at least 85% sequence identity to SEQ ID NO: 46 or SEQ ID NO: 61; LC-CDR3 having amino acids with at least 85% sequence identity to SEQ ID NO: 47 or SEQ ID NO: 62.

[0382] This has the advantage of being able to neutralize variants BQ.1.1 and XBB and may be used to treat subjects in need of treatment. Antibody 1, Antibody 2, and Antibody 5 were all able to effectively neutralize both SARS-CoV-2 variants BQ.1.1 and XBB at IC50 concentrations below 200 (see Figure 1E, Figure 1F, Figure 2A, and Figure 3). This demonstrates a significant improvement over the much-needed antibody E7, which requires even higher concentrations to inhibit variants BQ.1.1 and XBB, and the bebuterovimab (LY-CoV1404) antibody does not bind to these variants. In various embodiments, a concentration of E7 less than the concentration required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB refers to a concentration that is at least half the concentration of E7, or at least 3, 4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 29, 30, 34, 35, or 40 times lower than the concentration of E7.

[0383] In various embodiments, Antibody 1 comprises a VH region incorporating the following CDRs: HC-CDR1 having the amino acids of SEQ ID NO: 37; HC-CDR2 having the amino acids of SEQ ID NO: 38; and HC-CDR3 having the amino acids of SEQ ID NO: 39; and a VL region incorporating the following CDRs: LC-CDR1 having the amino acids of SEQ ID NO: 45; LC-CDR2 having the amino acids of SEQ ID NO: 46; and LC-CDR3 having the amino acids of SEQ ID NO: 47. In various embodiments, Antibody 1 comprises a heavy chain having the amino acids of SEQ ID NO: 36; and a light chain having the amino acids of SEQ ID NO: 44. Antibody 1 requires only 16.5 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, which is approximately 34-35 fold lower than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant BQ.1.1. Antibody 1 required only 11.6 ng / ml to effectively neutralize SARS-CoV-2 variant XBB, which is approximately 29- to 30-fold lower than the concentration of E7 required to bind and neutralize SARS-CoV-2 variant XBB.

[0384] In various embodiments, antibody 2 comprises a VH region incorporating the following CDRs: HC-CDR1 having the amino acids of SEQ ID NO: 37; HC-CDR2 having the amino acids of SEQ ID NO: 53; and HC-CDR3 having the amino acids of SEQ ID NO: 54, and a VL region incorporating the following CDRs: LC-CDR1 having the amino acids of SEQ ID NO: 60; LC-CDR2 having the amino acids of SEQ ID NO: 61; and LC-CDR3 having the amino acids of SEQ ID NO: 62. In various embodiments, antibody 2 comprises a heavy chain having the amino acids of SEQ ID NO: 52; and a light chain having the amino acids of SEQ ID NO: 59. Antibody 2 requires only 16.6 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, which is approximately 34 to 35 times lower than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant BQ.1.1. Antibody 2 required only 8.5 ng / ml to effectively neutralize SARS-CoV-2 variant XBB, which is approximately 40-fold lower than the concentration of E7 required to bind and neutralize SARS-CoV-2 variant XBB.

[0385] In various embodiments, antibody 5 comprises a heavy chain having the amino acid sequence of SEQ ID NO: 96; and a light chain having the amino acid sequence of SEQ ID NO: 102. Antibody 5 requires only 57.5 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, which is approximately one-tenth the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant BQ.1.1. Antibody 5 requires only 40.8 ng / ml to effectively neutralize SARS-CoV-2 variant XBB, which is approximately one-eighth the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant XBB.

[0386] The amino acid sequence of SEQ ID NO:822 includes the heavy chain amino acids of both SEQ ID NO:36 and SEQ ID NO:52, and the sequence encompassing the CDRs of SEQ ID NOs:107, 108, 109, 113, 114, and 115. The amino acid sequence of SEQ ID NO:823 includes the light chain amino acids of both SEQ ID NO:44 and SEQ ID NO:59, and the sequence encompassing the CDRs of SEQ ID NOs:110, 111, 112, 116, 117, and 118. In various embodiments, the E7 antibody comprises a heavy chain having the amino acid sequence of SEQ ID NO:824 and a light chain having the amino acid sequence of SEQ ID NO:830. Any inhibition assay known in the art for determining inhibition of variants from binding to ACE2, for example, the 50% inhibitory concentration (IC50; ng / ml) of a monoclonal antibody in blocking cell entry using a pseudovirus neutralization test format, can be used to determine antibodies that can be used at concentrations less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB.

[0387] The even higher neutralizing potency of the disclosed antibodies will allow for lower dosages of the antigen-binding molecules to be used clinically, either as individual antigen-binding molecules or mixed into a cocktail of two or more antigen-binding molecules or antigen-binding molecules with two or more different antigen-binding domains.

[0388] In various embodiments, Antibody 1 comprises the following VH regions: HC-FR1-SEQ ID NO:40; HC-CDR1-SEQ ID NO:37; HC-FR2-SEQ ID NO:41; HC-CDR2-SEQ ID NO:38; HC-FR3-SEQ ID NO:42; HC-CDR3-SEQ ID NO:39; HC-FR4-SEQ ID NO:43; and the following VL regions: LC-FR1-SEQ ID NO:48; LC-CDR1-SEQ ID NO:45; LC-FR2-SEQ ID NO:49; LC-CDR2-SEQ ID NO:46; LC-FR3-SEQ ID NO:50; LC-CDR3-SEQ ID NO:47; LC-FR4-SEQ ID NO:51.

[0389] In various embodiments, antibody 2 comprises the following VH regions: HC-FR1-SEQ ID NO:55; HC-CDR1-SEQ ID NO:37; HC-FR2-SEQ ID NO:56; HC-CDR2-SEQ ID NO:53; HC-FR3-SEQ ID NO:57; HC-CDR3-SEQ ID NO:54; HC-FR4-SEQ ID NO:58; and the following VL regions: LC-FR1-SEQ ID NO:63; LC-CDR1-SEQ ID NO:60; LC-FR2-SEQ ID NO:64; LC-CDR2-SEQ ID NO:61; LC-FR3-SEQ ID NO:65; LC-CDR3-SEQ ID NO:62; LC-FR4-SEQ ID NO:66.

[0390] In various embodiments, the terms bind and neutralize can include 50% or greater inhibition or neutralization of binding between a sarbecovirus spike protein and ACE 2. In various embodiments, 50% or greater inhibition or neutralization of binding between a sarbecovirus spike protein and ACE 2 can be selected from one or more of at least 50%, 55%, 60%, 61%, 62%, 63%, 64%, 65%, 69%, 70%, 71%, 72%, 73%, 74%, 75%, 76%, 77%, 78%, 79%, 80%, 81%, 82%, 83%, 84%, 85%, 86%, 87%, 88%, 89%, 90%, 95%, or 99% inhibition or neutralization.

[0391] In various embodiments, the heavy chain has at least 96%, or 97%, or 98%, or 99%, or 100% sequence identity to the amino acids of SEQ ID NO:822 or SEQ ID NO:36 or SEQ ID NO:52 or SEQ ID NO:96; and the light chain has at least 96%, or 97%, or 98%, or 99%, or 100% sequence identity to the amino acids of SEQ ID NO:823 or SEQ ID NO:44 or SEQ ID NO:59 or SEQ ID NO:102.

[0392] According to various embodiments, there is an antigen binding molecule that binds to and neutralizes SARS-CoV-2 variant BQ.1.1 at a concentration of E7 that is less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant BQ.1.1; at least one other SARS-CoV-2 variant, and another sarbecovirus other than SARS-CoV-2, wherein the antigen binding molecule is selected from the group consisting of (i) SEQ ID NO:822, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:67, SEQ ID NO:83, SEQ ID NO:96, SEQ ID NO:105, SEQ ID NO: 120, SEQ ID NO: 136, SEQ ID NO: 149, SEQ ID NO: 179, SEQ ID NO: 193, SEQ ID NO: 206, SEQ ID NO: 220, SEQ ID NO: 285, SEQ ID NO: 299, SEQ ID NO: 312, SEQ ID NO: 325, SEQ ID NO: 336, SEQ ID NO: 362, SEQ ID NO: 368, SEQ ID NO: 381, SEQ ID NO: 393, SEQ ID NO: 436, SEQ ID NO: 464, SEQ ID NO: 496, SEQ ID NO: 508, SEQ ID NO: 522, SEQ ID NO: 535, SEQ ID NO: 547, SEQ ID NO: 559, SEQ ID NO: 584, SEQ ID NO: 592, SEQ ID NO: 600, a heavy chain having at least 95% sequence identity to an amino acid sequence selected from any one of SEQ ID NO:614, SEQ ID NO:627, SEQ ID NO:643, and SEQ ID NO:656; and (ii) SEQ ID NO:823, SEQ ID NO:44, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:89, SEQ ID NO:102, SEQ ID NO:113, SEQ ID NO:128, SEQ ID NO:144, SEQ ID NO:157, SEQ ID NO:187, SEQ ID NO:200, SEQ ID NO:213, SEQ ID NO:228, SEQ ID NO:291, SEQ ID NO:305, SEQ ID NO:318, and a light chain having at least 95% sequence identity to an amino acid sequence selected from any one of SEQ ID NO:331, SEQ ID NO:343, SEQ ID NO:366, SEQ ID NO:374, SEQ ID NO:388, SEQ ID NO:400, SEQ ID NO:443, SEQ ID NO:471, SEQ ID NO:502, SEQ ID NO:515, SEQ ID NO:529, SEQ ID NO:542, SEQ ID NO:554, SEQ ID NO:567, SEQ ID NO:587, SEQ ID NO:596, SEQ ID NO:607, SEQ ID NO:621, SEQ ID NO:635, SEQ ID NO:651, and SEQ ID NO:663.

[0393] This has the advantage of being able to neutralize the variant BQ.1.1 in the ongoing pandemic and be used at concentrations suitable for treating subjects in need (see, e.g., [Figure 1E], [Figure 2A], and [Figure 3]). Antibodies 1 through 5, including Antibody 1, Antibody 2, Antibody 3, Antibody 4, and Antibody 5, were all able to effectively neutralize SARS-CoV-2 variant BQ.1.1 at IC50 concentrations of 300 ng / ml or less (Figure 3). This demonstrates a significant improvement over antibody E7, which requires much higher concentrations to inhibit this variant and the bebuterovimab (LY-CoV1404) antibody does not bind to this variant BQ.1.1. However, it also has the advantage of potentially helping to manage future outbreaks of other zoonotic sarbecovirus infections, such as those currently observed in bats or pangolins (see [Figure 3]).

[0394] In various embodiments, a concentration of E7 less than the concentration required to bind and neutralize SARS-CoV-2 variant BQ.1.1 refers to at least 1.1-fold the concentration of E7, or at least 1.1, 1.5, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 15, 20, 29, 30, 34, 35, or 40-fold the concentration of E7. Antibody 1 requires only 16.5 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, which is about 34-35 fold lower than the concentration of E7 required to bind and neutralize SARS-CoV-2 variant BQ.1.1. Antibody 2 requires only 16.6 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, which is about 34-35 fold lower than the concentration of E7 required to bind and neutralize SARS-CoV-2 variant BQ.1.1. Antibody 3 required only 270.6 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, approximately half the concentration of E7 required to bind and neutralize SARS-CoV-2 variant BQ.1.1. Antibody 4 required only 48.4 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, approximately 11-fold lower than the concentration of E7 required to bind and neutralize SARS-CoV-2 variant BQ.1.1. Antibody 5 required only 57.5 ng / ml to effectively neutralize SARS-CoV-2 variant BQ.1.1, approximately 10-fold lower than the concentration of E7 required to bind and neutralize SARS-CoV-2 variant BQ.1.1. Antibodies 6–9, 11–14, 19–23, 25–28, 32, 35, 39–44, and 46–52 were also all able to effectively neutralize SARS-CoV-2 variant BQ.1.1 with IC50 concentrations below 550 ng / ml.

[0395] In various embodiments, the above sequences refer to antibodies 1-5, 6-9, 11-14, 19-23, 25-28, 32, 35, 39-44, and 46-52 listed in Table C herein. In contrast, antibodies 10, 15-18, 24, 29-31, 33, 34, 36-38, and 45 were able to bind and neutralize SARS-CoV-2 variant BQ.1.1 in some cases, but were less effective than the E7 antibody, thereby requiring higher concentrations than E7 to bind and neutralize SARS-CoV-2 variant BQ.1.1 (see Figure 3).

[0396] In various embodiments, the at least one other SARS-CoV-2 variant includes the alpha COVID-19 variant SARS-CoV-2 B.1.1.7; the beta COVID-19 variant SARS-CoV-2 B.1.351, also known as the 20H / 501Y.V2 or 501Y.V2 variant; the gamma variant P.1, the delta SARS-CoV-2 B.1.617.2; and any one of the omicron variants SARS-CoV-2 B.1.1.529 BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, and XBB. In various embodiments, the at least one other SARS-CoV-2 variant can include two or more, three or more, four or more, five or more, six or more, or seven or more of any one of the following: alpha COVID-19 variant SARS-CoV-2 B.1.1.7; beta COVID-19 variant SARS-CoV-2 B.1.351, also known as the 20H / 501Y.V2 or 501Y.V2 variant; gamma variant P.1; delta SARS-CoV-2 B.1.617.2; and omicron variant SARS-CoV-2 B.1.1.529 BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, and XBB. As used herein, the term "another sarbecovirus" refers to a sarbecovirus that is not SARS-CoV-2. In various embodiments, the other sarbecovirus includes SARS-CoV, BANAL-52, WIV-1, SC2r-CoV RaTG13, SC2r-CoV GX-P5L, SC2r-CoV GD-1, SC2r-CoV mYN02, RacCS203, or a future unknown sarbecovirus. In various embodiments, the other sarbecovirus includes any betacoronavirus that uses the ACE2 receptor to enter cells and is not SARS-CoV-2 but is a sarbecovirus other than SARS-CoV-2. A broad-spectrum antigen-binding molecule has the advantage of being able to block most sarbecoviruses, effectively helping to prevent infection with both known and unknown sarbecoviruses.In various embodiments, the antigen-binding molecule comprises a monoclonal antibody (mAb), which may be one of the most efficient and powerful tools for rapid development and deployment in the fight against future emergence of animal-borne viruses, and particularly sarbecoviruses.

[0397] In various embodiments, there is a composition of any one of the antigen-binding molecules described herein above, bebuterovimab LY-CoV1404, and E7. In various embodiments, the bebuterovimab LY-CoV1404 antibody includes those described in WO / 2021 / 183359. In various embodiments, such compositions or cocktails have the advantage of increasing the range of antigen-binding molecules that bind and neutralize a wide range of SARS-CoV-2 variants and other sarbecoviruses (see, e.g., Figure 3). In various embodiments, the composition comprises: (i) a VH region incorporating the following CDRs: HC-CDR1 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 37; HC-CDR2 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 53; HC-CDR3 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 39 or SEQ ID NO: 54; and (ii) a VL region incorporating the following CDRs: LC-CDR1 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 45 or SEQ ID NO: 60; LC-CDR2 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 46 or SEQ ID NO: 61; LC-CDR3 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 47 or SEQ ID NO: 62; and (iii) a heavy chain comprising amino acids having at least 95% sequence identity to the sequence of SEQ ID NO: 824; and (iv) a light chain comprising amino acids having at least 95% sequence identity to the sequence of SEQ ID NO: 830. A second antigen-binding molecule comprising: Includes.

[0398] In various embodiments, the composition comprises an antigen binding molecule selected from any one of antibodies 1-52 in combination with the antibody bebuterovimab LY-CoV1404 or antibody E7.

[0399] According to various embodiments, there is a method of treating a sarbecovirus infection, the method comprising administering to a patient in need thereof a therapeutically effective amount of an antigen binding molecule or composition described herein. In various embodiments, the therapeutically effective amount of the described antigen binding molecule or composition comprises an amount capable of neutralizing or inhibiting a sarbecovirus infection at least sufficiently to stop, minimize, or reduce symptoms of the sarbecovirus infection. According to various embodiments, there is a method of treating a sarbecovirus infection, the method comprising administering to a patient in need thereof a therapeutically effective amount of an antigen binding molecule that binds to and neutralizes SARS-CoV-2 variants BQ.1.1 and XBB at a concentration less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB, wherein the antigen binding molecule has (i) the following CDRs: HC-CDR1 having an amino acid sequence having at least 85% sequence identity to SEQ ID NO: 37; and (ii) a VH region incorporating the following CDRs: an HC-CDR2 having an amino acid sequence that has at least 85% sequence identity to SEQ ID NO: 39 or SEQ ID NO: 54; an HC-CDR3 having an amino acid sequence that has at least 85% sequence identity to SEQ ID NO: 45 or SEQ ID NO: 60; an LC-CDR2 having an amino acid sequence that has at least 85% sequence identity to SEQ ID NO: 46 or SEQ ID NO: 61; and an LC-CDR3 having an amino acid sequence that has at least 85% sequence identity to SEQ ID NO: 47 or SEQ ID NO: 62.

[0400] According to various embodiments, there is a method of treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of an antigen binding molecule that binds to and neutralizes SARS-CoV-2 variants BQ.1.1 and XBB at a concentration less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB, wherein the antigen binding molecule comprises: (i) a heavy chain having amino acids with at least 95% sequence identity to SEQ ID NO:822 or SEQ ID NO:36 or SEQ ID NO:52 or SEQ ID NO:96; and (ii) a light chain having amino acids with at least 95% sequence identity to SEQ ID NO:823 or SEQ ID NO:44 or SEQ ID NO:59 or SEQ ID NO:102.

[0401] According to various embodiments, there is provided a method of treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of an antigen binding molecule that binds to and neutralizes SARS-CoV-2 variant BQ.1.1 at a concentration less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant BQ.1.1; at least one other SARS-CoV-2 variant and another sarbecovirus, wherein the antigen binding molecule is selected from the group consisting of (i) SEQ ID NO: 822, SEQ ID NO: 36, SEQ ID NO: 52 , SEQ ID NO: 67, SEQ ID NO: 83, SEQ ID NO: 96, SEQ ID NO: 105, SEQ ID NO: 120, SEQ ID NO: 136, SEQ ID NO: 149, , SEQ ID NO: 179, SEQ ID NO: 193, SEQ ID NO: 206, SEQ ID NO: 220, SEQ ID NO: 285, SEQ ID NO: 299, SEQ ID NO: 312, SEQ ID NO: 325, SEQ ID NO: 336, SEQ ID NO: 362, SEQ ID NO: 368, SEQ ID NO: 381, SEQ ID NO: 393, SEQ ID NO: 436, SEQ ID NO: 464, SEQ ID NO: 496, SEQ ID NO: 508, SEQ ID NO: 522, SEQ ID NO: 535, SEQ ID NO: 547, SEQ ID NO: 559, SEQ ID NO: SEQ ID NO:584, SEQ ID NO:592, SEQ ID NO:600, SEQ ID NO:614, SEQ ID NO:627, SEQ ID NO:643, and SEQ ID NO:656; and (ii) a heavy chain having at least 95% sequence identity to an amino acid selected from any one of SEQ ID NO:823, SEQ ID NO:44, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:89, SEQ ID NO:102, SEQ ID NO:113, SEQ ID NO:128, SEQ ID NO:144, SEQ ID NO:157, SEQ ID NO:187, SEQ ID NO:200, SEQ ID NO:213, SEQ ID NO:228, SEQ ID NO:291, SEQ ID NO:3 05, SEQ ID NO:318, SEQ ID NO:331, SEQ ID NO:343, SEQ ID NO:366, SEQ ID NO:374, SEQ ID NO:388, SEQ ID NO:400, SEQ ID NO:443, SEQ ID NO:471, SEQ ID NO:502, SEQ ID NO:515, SEQ ID NO:529, SEQ ID NO:542, SEQ ID NO:554, SEQ ID NO:567, SEQ ID NO:587, SEQ ID NO:596, SEQ ID NO:607, SEQ ID NO:621, SEQ ID NO:635, SEQ ID NO:651 and SEQ ID NO:663.

[0402] According to various embodiments, there is a method for treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of a composition comprising an antigen-binding molecule described herein and any one of bebuterovimab LY-CoV1404 and E7. A composition comprising a combination of antibody 1 and antibody E7 was able to enhance inhibitory potency, resulting in effective inhibition of all tested SARS-CoV-2 variants and all tested sarbecoviruses. Similarly, a composition comprising a combination of antibody 2 and antibody E7 was able to enhance inhibitory potency, resulting in effective inhibition of all tested SARS-CoV-2 variants and all tested sarbecoviruses.

[0403] In various embodiments, the patient in need can be an individual diagnosed with a sarbecovirus infection. In various embodiments, the patient in need can be an individual diagnosed with an infection caused by a SARS-CoV-2 variant. In various embodiments, the method includes determining the infection caused by a sarbecovirus, e.g., a SARS-CoV-2 variant. In various embodiments, the patient in need can be an individual diagnosed with COVID-19 caused by a sarbecovirus, e.g., a SARS-CoV-2 variant. In various embodiments, the patient in need can be an individual diagnosed with COVID-19 caused by a SARS-CoV-2 variant selected from BQ.1.1 and XBB.

[0404] According to various embodiments, there is provided an antigen binding molecule or composition described herein, or an effective amount of the composition described herein, for use in treating a sarbecovirus infection. In various embodiments, the sarbecovirus infection may be caused by a non-SARS-CoV-2, such as a sarbecovirus previously known to infect bats or pangolins, or any currently unknown sarbecovirus. In various embodiments, the sarbecovirus infection may be caused by a SARS-CoV-2 variant. In various embodiments, the sarbecovirus infection may be caused by a SARS-CoV-2 variant selected from BQ.1.1 and XBB. In various embodiments, the antigen binding molecules discussed herein are suitable for use in treating individuals diagnosed with a sarbecovirus infection.

[0405] The following numbered sections describe certain aspects and embodiments of the present disclosure: 1. An antigen-binding molecule that binds to and neutralizes SARS-CoV-2 variants BQ.1.1 and XBB at a concentration of E7 that is less than the concentration required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB, comprising a heavy chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 822 or SEQ ID NO: 36 or SEQ ID NO: 52 or SEQ ID NO: 96; and a light chain having an amino acid sequence with at least 95% sequence identity to SEQ ID NO: 823 or SEQ ID NO: 44 or SEQ ID NO: 59 or SEQ ID NO: 102. 2. An antigen-binding molecule according to item 1, wherein (i) the heavy chain variable (VH) region incorporates the following CDRs: HC-CDR1 having amino acids having at least 85% sequence identity to SEQ ID NO: 37; HC-CDR2 having amino acids having at least 85% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 53; HC-CDR3 having amino acids having at least 85% sequence identity to SEQ ID NO: 39 or SEQ ID NO: 54; and (ii) the light chain variable (VL) region incorporates the following CDRs: LC-CDR1 having amino acids having at least 85% sequence identity to SEQ ID NO: 45 or SEQ ID NO: 60; LC-CDR2 having amino acids having at least 85% sequence identity to SEQ ID NO: 46 or SEQ ID NO: 61; LC-CDR3 having amino acids having at least 85% sequence identity to SEQ ID NO: 47 or SEQ ID NO: 62. 3. SARS-CoV-2 variant BQ.1.1; An antigen-binding molecule that binds to and neutralizes SARS-CoV-2 variant BQ.1.1 at a concentration of E7 that is less than the concentration of E7 required to bind to and neutralize at least one other SARS-CoV-2 variant and another sarbecovirus, comprising: (i) SEQ ID NO:822, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:67, SEQ ID NO:83, SEQ ID NO:96, SEQ ID NO:105, SEQ ID NO:120, SEQ ID NO:136, SEQ ID NO:149, SEQ ID NO:150, SEQ ID NO:160, SEQ ID NO:172, SEQ ID NO:174, SEQ ID NO:176, SEQ ID NO:178, SEQ ID NO:179, SEQ ID NO:180, SEQ ID NO:181, SEQ ID NO:182, SEQ ID NO:183, SEQ ID NO:184, SEQ ID NO:185, SEQ ID NO:186, SEQ ID NO:187, SEQ ID NO:188, SEQ ID NO:189, SEQ ID NO:190, SEQ ID NO:192, SEQ ID NO:193, SEQ ID NO:194, SEQ ID NO:195, SEQ ID NO:196, SEQ ID NO:197, SEQ ID NO:198, SEQ ID NO:200, SEQ ID NO:201, SEQ ID NO:202, SEQ ID NO:203, SEQ ID NO:204, SEQ ID NO:205, SEQ ID NO:206, SEQ ID NO:207, SEQ ID NO:208, SEQ ID NO:209, SEQ ID NO:210, SEQ ID NO:211, S Column number 179, SEQ ID NO:193, SEQ ID NO:206, SEQ ID NO:220, SEQ ID NO:285, SEQ ID NO:299, SEQ ID NO:312, SEQ ID NO:325, SEQ ID NO:336, SEQ ID NO:362, SEQ ID NO:368, SEQ ID NO:381, SEQ ID NO:393, SEQ ID NO:436, SEQ ID NO:464, SEQ ID NO:496, SEQ ID NO:508, SEQ ID NO:522, SEQ ID NO:535, SEQ ID NO:547, SEQ ID NO:559, SEQ ID NO:584, SEQ ID NO:592, SEQ ID NO:600, SEQ ID NO:614, SEQ ID NO:627, SEQ ID NO: and (ii) a light chain having at least 95% sequence identity to an amino acid selected from any one of SEQ ID NOs: 823, 44, 59, 75, 89, 102, 113, 128, 144, 157, 187, 200, 213, 228, 291, 305, 318, 331, 343, 366, 374, 388, 400, 443, 471, 502, 515, 529, 542, 554, 567, 587, 596, 607, 621, 635, 651, and 663. An antigen-binding molecule comprising: 4. A composition comprising the antigen-binding molecule of any one of paragraphs 1 to 3 and any one of bebuterovimab LY-CoV1404 and E7. 5. The antigen-binding molecule comprises: (i) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 37; HC-CDR2 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 53; HC-CDR3 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 39 or SEQ ID NO: 54; and (ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 45 or SEQ ID NO: 60; LC-CDR2 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 46 or SEQ ID NO: 61; LC-CDR3 comprising amino acids having at least 85% sequence identity to SEQ ID NO: 47 or SEQ ID NO: 62, wherein E7 comprises (iii) a heavy chain comprising an amino acid sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 824; and (iv) a light chain comprising an amino acid sequence having at least 95% sequence identity to the sequence set forth in SEQ ID NO: 830. The composition according to paragraph 4, comprising a second antigen-binding molecule. 6. A composition that binds to and neutralizes SARS-CoV-2 variant BQ.1.1, at least three other SARS-CoV-2 variants, and another sarbecovirus, comprising: (i) SEQ ID NO:822, SEQ ID NO:36, SEQ ID NO:52, SEQ ID NO:67, SEQ ID NO:83, SEQ ID NO:96, SEQ ID NO:105, SEQ ID NO:120, SEQ ID NO:136, SEQ ID NO:149, SEQ ID NO:164, SEQ ID NO:179, SEQ ID NO:193, SEQ ID NO:206, SEQ ID NO:220, SEQ ID NO:235, SEQ ID NO:249, SEQ ID NO:262, SEQ ID NO:274, SEQ ID NO:285, SEQ ID NO:299, SEQ ID NO:3 12, any one of SEQ ID NO:325, SEQ ID NO:336, SEQ ID NO:350, SEQ ID NO:362, SEQ ID NO:368, SEQ ID NO:381, SEQ ID NO:393, SEQ ID NO:405, SEQ ID NO:416, SEQ ID NO:427, SEQ ID NO:436, SEQ ID NO:449, SEQ ID NO:453, SEQ ID NO:464, SEQ ID NO:475, SEQ ID NO:487, SEQ ID NO:496, SEQ ID NO:508, SEQ ID NO:522, SEQ ID NO:535, SEQ ID NO:547, SEQ ID NO:559, SEQ ID NO:572, SEQ ID NO:584, SEQ ID NO:592, SEQ ID NO:600, SEQ ID NO:614, SEQ ID NO:627, SEQ ID NO:643 and SEQ ID NO:656 and (ii) a heavy chain having at least 95% sequence identity to an amino acid sequence selected from SEQ ID NO:823, SEQ ID NO:44, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:89, SEQ ID NO:102, SEQ ID NO:113, SEQ ID NO:128, SEQ ID NO:144, SEQ ID NO:157, SEQ ID NO:171, SEQ ID NO:187, SEQ ID NO:200, SEQ ID NO:213, SEQ ID NO:228, SEQ ID NO:242, SEQ ID NO:256, SEQ ID NO:270, SEQ ID NO:280, SEQ ID NO:291, SEQ ID NO:305, SEQ ID NO:318, SEQ ID NO:331, SEQ ID NO:343, SEQ ID NO:355, SEQ ID NO:366, a light chain having at least 95% sequence identity to amino acids selected from any one of SEQ ID NO:374, SEQ ID NO:388, SEQ ID NO:400, SEQ ID NO:411, SEQ ID NO:423, SEQ ID NO:432, SEQ ID NO:443, SEQ ID NO:451, SEQ ID NO:460, SEQ ID NO:471, SEQ ID NO:481, SEQ ID NO:491, SEQ ID NO:502, SEQ ID NO:515, SEQ ID NO:529, SEQ ID NO:542, SEQ ID NO:554, SEQ ID NO:567, SEQ ID NO:579, SEQ ID NO:587, SEQ ID NO:596, SEQ ID NO:607, SEQ ID NO:621, SEQ ID NO:635, SEQ ID NO:651, and SEQ ID NO:663;and a composition comprising an antigen-binding molecule comprising bepterovimab LY-CoV1404 and a second antigen-binding molecule comprising any one of E7; 6. A method of treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of an antigen-binding molecule that binds to and neutralizes SARS-CoV-2 variants BQ.1.1 and XBB at a concentration less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB, wherein the antigen-binding molecule is (i) a heavy chain variable (VH) region incorporating the following CDRs: HC-CDR1 having amino acids with at least 85% sequence identity to SEQ ID NO: 37; HC-CDR2 having amino acids with at least 85% sequence identity to SEQ ID NO: 38 or SEQ ID NO: 53; HC-CDR3 having amino acids with at least 85% sequence identity to SEQ ID NO: 39 or SEQ ID NO: 54; and (ii) a light chain variable (VL) region incorporating the following CDRs: LC-CDR1 having amino acids with at least 85% sequence identity to SEQ ID NO: 45 or SEQ ID NO: 60; LC-CDR2 having amino acids with at least 85% sequence identity to SEQ ID NO: 46 or SEQ ID NO: 61; LC-CDR3 having amino acids with at least 85% sequence identity to SEQ ID NO: 47 or SEQ ID NO: 62. A method comprising: 7. A method for treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of an antigen binding molecule that binds to and neutralizes SARS-CoV-2 variants BQ.1.1 and XBB at a concentration less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variants BQ.1.1 and XBB, wherein the antigen binding molecule comprises: (i) a heavy chain having amino acids with at least 95% sequence identity to SEQ ID NO: 822 or SEQ ID NO: 36 or SEQ ID NO: 52 or SEQ ID NO: 96; and (ii) a light chain having amino acids with at least 95% sequence identity to SEQ ID NO: 823 or SEQ ID NO: 44 or SEQ ID NO: 59 or SEQ ID NO: 102. 8. A method of treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of an antigen-binding molecule that binds to and neutralizes SARS-CoV-2 variant BQ.1.1 at a concentration less than the concentration of E7 required to bind to and neutralize SARS-CoV-2 variant BQ.1.1; at least one other SARS-CoV-2 variant and another sarbecovirus, wherein the antigen-binding molecule is selected from the group consisting of (i) SEQ ID NO: 822, SEQ ID NO: 36, SEQ ID NO: 52, SEQ ID NO: 67, SEQ ID NO: 83, SEQ ID NO:96, SEQ ID NO:105, SEQ ID NO:120, SEQ ID NO:136, SEQ ID NO:149, SEQ ID NO:179, SEQ ID NO:193, SEQ ID NO:206, SEQ ID NO:220, SEQ ID NO:285, SEQ ID NO:299, SEQ ID NO:312, SEQ ID NO:325, SEQ ID NO:336, SEQ ID NO:362, SEQ ID NO:368, SEQ ID NO:381, SEQ ID NO:393, SEQ ID NO:436, SEQ ID NO:464, SEQ ID NO:496, SEQ ID NO:508, SEQ ID NO:522, SEQ ID NO:535, SEQ ID NO:547, SEQ ID NO:559, SEQ ID NO:584, SEQ ID NO:5 92, SEQ ID NO:600, SEQ ID NO:614, SEQ ID NO:627, SEQ ID NO:643, and SEQ ID NO:656; and (ii) a heavy chain having at least 95% sequence identity to an amino acid selected from any one of SEQ ID NO:823, SEQ ID NO:44, SEQ ID NO:59, SEQ ID NO:75, SEQ ID NO:89, SEQ ID NO:102, SEQ ID NO:113, SEQ ID NO:128, SEQ ID NO:144, SEQ ID NO:157, SEQ ID NO:187, SEQ ID NO:200, SEQ ID NO:213, SEQ ID NO:228, SEQ ID NO:291, SEQ ID NO:305, SEQ ID NO: SEQ ID NO:318, SEQ ID NO:331, SEQ ID NO:343, SEQ ID NO:366, SEQ ID NO:374, SEQ ID NO:388, SEQ ID NO:400, SEQ ID NO:443, SEQ ID NO:471, SEQ ID NO:502, SEQ ID NO:515, SEQ ID NO:529, SEQ ID NO:542, SEQ ID NO:554, SEQ ID NO:567, SEQ ID NO:587, SEQ ID NO:596, SEQ ID NO:607, SEQ ID NO:621, SEQ ID NO:635, SEQ ID NO:651 and SEQ ID NO:663. 9. A method for treating a sarbecovirus infection, comprising administering to a patient in need thereof a therapeutically effective amount of a composition comprising the antigen-binding molecule of paragraph 1 or 3 and any one of bebuterovimab LY-CoV1404 and E7. 10. A therapeutically effective amount of an antigen-binding molecule according to any one of paragraphs 1 to 3 or a composition according to paragraph 4 or 5 for use in treating a sarbecovirus infection. 11. The antigen-binding molecule or composition for use in paragraph 10, wherein the sarbecovirus infection is caused by a SARS-CoV-2 variant, including BQ.1.1. 12. A therapeutically effective amount of an antigen-binding molecule or composition for use according to paragraph 10, wherein the sarbecovirus infection is caused by a SARS-CoV-2 variant including BQ.1.1 or XBB. *** The present disclosure includes combinations of the described embodiments and preferred features unless such combinations are expressly not permitted or expressly avoided.

[0406] The section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described. Aspects and embodiments of the present disclosure are illustrated herein, by way of example, with reference to the accompanying figures. Further aspects and embodiments will be apparent to those skilled in the art. All documents mentioned in this specification are incorporated herein by reference.

[0407] Throughout this specification, including the claims which follow, unless the context requires otherwise, the word "comprise" and variations such as "comprises" and "comprising" will be understood to imply the inclusion of a stated integer or step or group of integers or steps, but not the exclusion of any other integer or step or group of integers or steps.

[0408] As used herein, an amino acid sequence or region of a polypeptide that "corresponds" to a particular reference amino acid sequence or region of a polypeptide has at least 60%, for example, at least 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% sequence identity to the amino acid sequence of the amino acid sequence / polypeptide / region. An amino acid sequence / region / position of a polypeptide / amino acid sequence that "corresponds" to a particular reference amino acid sequence / region / position of a polypeptide / amino acid sequence can be identified by sequence alignment of the subject sequence to the reference sequence, for example, using sequence alignment software such as ClustalOmega (Soding, J. 2005, Bioinformatics 21, 951-960).

[0409] It should be noted that, as used in the specification and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Ranges may be expressed herein as from "about" one particular value and / or to "about" another particular value. When such a range is expressed, another embodiment includes from the one particular value and / or to the other particular value. Similarly, when values ​​are expressed as approximately, by use of the antecedent "about," it will be understood that the particular value forms another embodiment.

[0410] When a nucleic acid sequence is disclosed herein, its reverse complement is also expressly contemplated. The methods described herein may preferably be performed in vitro. The term "in vitro" is intended to encompass procedures performed with cells in culture, while the term "in vivo" is intended to encompass procedures using / on intact multicellular organisms. [Example]

[0411] Example 1 material and method 1.1 Pseudovirus production Vesicular stomatitis virus (VSV) pseudotyped with full-length SARS-CoV-2 Wuhan-hu-1 (ancestral), Omicron BA.1, Omicron BA.2, Omicron BA.5, Omicron BA.2.75, Omicron BA.2.75.2, Omicron BA.4.6.1, Omicron BF.7, Omicron BQ.1.1, Omicron XBB.1, Omicron XBB.1.16, Omicron XBB.2.3, Omicron EG.5, Omicron EG5.1, BANAL-52, GD-1, GX-P5L, WIV-1, or SARS-CoV-1 spike proteins were produced and packaged as described in Tan et al., Nat. Biotechnol. (2020) 38:1073–1078, with minor modifications. Briefly, 5 million HEK293T cells were transfected with 20 μg of pCAGGS plasmid encoding the relevant spike protein using FuGENE6 (Promega). 24 h posttransfection, cells were incubated with VSVΔG luc seed virus (at an MOI of 5) for 2 h. After washing twice with phosphate-buffered saline (PBS), infected cells were replenished with complete growth medium supplemented with a 1:5000 dilution of anti-VSV-G mAb (clone 8GF11, Kerafast). 24 h postinfection, pseudoviruses were harvested by centrifugation at 2,000 x g for 5 min.

[0412] 1.2 Pseudovirus Neutralization Test (pVNT) For pVNT, 3x10 6RLU pseudoviruses were preincubated with serially diluted monoclonal antibodies in PBS buffer. For serum-spiked pVNT, the buffer was supplemented with human serum at a 1:20 dilution to mimic ex vivo conditions. The starting concentration of the monoclonal antibodies was 20 μg / ml, serially diluted 4-fold in a final volume of 50 μL for 1 h at 37°C. This was followed by infection of A549 cells stably expressing human ACE2 with the pseudovirus-mAb mixture. 20–24 h after infection, an equal volume of ONE-Glo luciferase substrate (Promega) was added, and the luminescence signal was measured using a Cytation5 microplate reader (BioTek) with Gen5 software, version 3.10.

[0413] 1.3 Multiplex Surrogate Virus Neutralization Test (sVNT) A multiplex surrogate virus neutralization test (sVNT) was performed essentially as described in Tan et al., Nat. Biotechnol. (2020) 38:1073-1078, using receptor binding domain (RBD) proteins from 11 different sarbecoviruses: SARS-CoV-2; SARS-CoV-2 B.1.351 (beta); SARS-CoV-2 B.1.617.2 (delta); SARS-CoV-2 B.1.1.529.1 (BA.1); SARS-CoV-2 B.1.1.529.5 (BA.5); SARS-CoV-2 XBB.1; SC1r-CoV Rs2018B; SC1r-CoV RsSHC014; SARS-CoV and Bat-CoV Khosta-2.

[0414] Briefly, AviTag-biotinylated RBDs from various sarbecoviruses were coated onto MagPlex Avidin microspheres (Luminex) at 5 μg per million beads. The RBD-coated microspheres (600 beads / antigen) were preincubated with test monoclonal antibodies at a starting concentration of 10 μg / mL in four-fold serial dilutions for 15 minutes at 37°C with 250 rpm agitation. After the 15-minute incubation, 50 μL of 2 μg / mL phycoerythrin (PE)-conjugated hACE2 (GenScript) was added to the wells and incubated for 15 minutes at 37°C with agitation, followed by two PBS-1% bovine serum albumin washes. Data were acquired using the MAGPIX (Luminex) system.

[0415] Example 2 Analysis of the Ability of Antibodies to Neutralize Infection of ACE2-Expressing Cells by Pseudosarbecoviruses Expressing Sarbecovirus Spike Proteins Antibodies capable of binding to the SARS-CoV-2 spike protein were obtained, and their sequence characteristics are summarized in Tables A to C.

[0416] The ability of various antibodies to neutralize infection of human ACE2-expressing cells by pseudoviruses expressing the spike proteins of SARS-CoV-2 and six SARS-CoV-2 variants (BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, and XBB) was analyzed using a pseudovirus neutralization test (pVNT). Fifty-one different antibodies were shown to bind to BQ.1.1 and / or other SARS-CoV-2 variants and to be able to neutralize infection by BQ.1.1 and / or other SARS-CoV-2 variants.

[0417] Figures 1A to 1F show results obtained in pVNT for two known antibodies against the SARS-CoV-2 spike protein (LyCoV-1404 and E7) and Ab1, Ab2, Ab3, Ab4, and Ab38. Ab1, Ab2, Ab3, and Ab4 were found to inhibit infection of ACE2-expressing cells by pseudoviruses expressing the spike protein of SARS-CoV-2 and all SARS-CoV-2 variants tested.

[0418] Ab1, Ab2, Ab3, Ab4, and Ab38 neutralized infection of ACE2-expressing cells by pseudoviruses expressing the BQ.1.1 spike protein, whose infection was not inhibited by LyCoV-1404 (Figure 1E). Furthermore, Ab1, Ab2, Ab3, and Ab4 were even more potent than E7 in inhibiting infection of ACE2-expressing cells by pseudoviruses expressing the BQ.1.1 spike protein.

[0419] Ab1, Ab2, and Ab3 neutralized infection of ACE2-expressing cells by pseudoviruses expressing the XBB.1 spike protein, whose infection was not inhibited by LyCoV-1404 (Figure 1F). Furthermore, Ab1 and Ab2 were even more potent than E7 in inhibiting infection of ACE2-expressing cells by pseudoviruses expressing the XBB.1 spike protein.

[0420] Furthermore, Ab1 and Ab2 inhibited infection of ACE2-expressing cells by pseudoviruses expressing the SARS-CoV-2, BA2.75.2, BF.7, or BA.4.6.1 spike proteins with potency similar to that of LyCoV-1404 and improved compared to E7.

[0421] The antibodies were then evaluated in pVNT against pseudoviruses expressing spike proteins from a wide range of sarbecoviruses, including SARS-CoV-2, SARS-CoV-2 variants BA.1, BA.2, BA.5, BA.2.75, BA.2.75.2, BA.4.6.1, BF.7, BQ.1.1, and XBB, clade 1b sarbecovirus BANAL-20-52 (BANAL-52), GD-1, GX-P5L, and clade 1a sarbecovirus WIV-1, as well as SARS-CoV-1. We also investigated the performance of certain combinations of antibodies (E7 + LyCoV-1404, E7 + Ab1, E7 + H12.2) at a 1:1 ratio. IC values ​​derived from % inhibition response curves fitted to the neutralization data were used. 50 The values ​​are shown in Figures 2A to 2C.

[0422] Numerous antibodies were determined to neutralize infection of ACE2-expressing cells by pseudoviruses expressing spike proteins from sarbecoviruses whose infection of ACE2-expressing cells was not inhibited by LyCoV-1404. Some antibodies inhibited infection of ACE2-expressing cells by pseudoviruses expressing spike proteins from sarbecoviruses with significantly greater potency than E7.

[0423] Ab1 (B11.2) and Ab2 (H12.2) demonstrated highly potent neutralization of 13 of 15 and 14 of 15 pseudosarbecoviruses (i.e., pseudoviruses expressing spike proteins from all clade 1b sarbecoviruses examined) tested, respectively, with IC50 values ​​ranging from 1.1 to 30.4 ng / mL for Ab1 and 0.3 to 16.6 ng / mL for Ab2 (shown in Figure 2A). Preparations containing E7 + Ab1 or E7 + Ab2 were able to neutralize all pseudosarbecoviruses (i.e., pseudoviruses expressing spike proteins from all clade 1a and 1b sarbecoviruses examined) tested, with IC50 values ​​ranging from 0.9 to 69.2 ng / mL.

[0424] The control antibody LyCoV-1404 showed strong activity against 10 of the 15 sarbecoviruses tested but was inactive against the remaining six, including SARS-CoV-2 variants BQ.1.1 or XBB.

[0425] The control antibody E7 demonstrated neutralizing capacity against all variants tested, but showed significantly lower potency compared to Ab1 and Ab2 against most SARS-CoV-2 variants tested, including BQ.1.1 and XBB.

[0426] These data demonstrate the broad and highly potent capabilities of the newly developed antibodies Ab1 and Ab2, including their ability to neutralize clade 1 sarbecoviruses, including the SARS-CoV-2 variants BQ.1.1 and XBB.1, which are not neutralized with high potency by the known antibodies E7 and LyCoV-1404.

[0427] Example 3 Analysis of the ability of antibodies to neutralize infection of ACE2-expressing cells by pseudosarbecoviruses expressing SARS-CoV-2 variants XBB.1.16, XBB.2.3, EG.5, or EG5.1 spike proteins We next investigated the ability of Ab2(H12.2) in additional pVNTs to inhibit infection of ACE2-expressing cells by pseudoviruses expressing the SARS-CoV-2 variants XBB.1.16, XBB.2.3, EG.5, or EG5.1 spike proteins. We also investigated its performance against these pseudosarbecoviruses, E7 and LyCoV1404.

[0428] The results are shown in Figure 3. LyCoV1404 was unable to neutralize pseudoviruses corresponding to these SARS-CoV-2 variants. Ab2 showed similar or improved ability to neutralize pseudoviruses corresponding to these SARS-CoV-2 variants compared to E7. In particular, Ab2 was highly potent in inhibiting infection of ACE2-expressing cells by pseudoviruses expressing the SARS-CoV-2 variant XBB.1.16 spike protein.

[0429] Example 4 Serum spike pVNT We next investigated the ability of Ab2(H12.2), E7, and the combination of Ab2+E7 (at a 1:1 ratio) to neutralize infection of ACE2-expressing cells by pseudoviruses expressing spike proteins of various sarbecoviruses in modified pVNTs, in which human serum was included in the reaction buffer (see Example 1.2).

[0430] The results are shown in Figure 4 and are consistent with those described in Examples 2 and 3. Ab2 showed similar or improved ability to neutralize infection of ACE2-expressing cells by all pseudosarbecoviruses expressing spike proteins from clade 1b sarbecoviruses compared to E7. Ab2 also showed similar ability to neutralize infection of ACE2-expressing cells by pseudosarbecoviruses expressing spike proteins from clade 1a sarbecovirus WIV-1 compared to E7. The combination of Ab2 and E7 potently neutralized all infection of ACE2-expressing cells by all pseudosarbecoviruses evaluated, including those expressing spike proteins from clade 1a sarbecoviruses (WIV-1 and SARS-CoV-1).

[0431] Example 5 Ab2 shows neutralization against clade 3 sarbecoviruses. We next investigated the ability of Ab2 and E7 to inhibit the interaction between ACE2 and a polypeptide consisting essentially of the RBD of the spike proteins of a variety of different sarbecoviruses in a multiplex surrogate virus neutralization assay (sVNT; see Example 1.3).

[0432] The experiment investigated the ability of antibodies to inhibit the interaction between the RBD of Khosta-2 and ACE2. Khosta-2 is a clade 3 sarbecovirus that exhibits binding to human ACE2 and is resistant to current SARS-CoV-2 vaccines, thereby representing a potential threat for future human infections (see, e.g., Seifert et al. PLoS Pathog. (2022) 18(9):e1010828).

[0433] The results are shown in Figure 5. Both Ab2 and E7 inhibited the interaction between the Khosta-2 RBD and human ACE2. E7 inhibited the interaction between ACE2 and the RBD of all sarbecoviruses tested, while Ab2 inhibited the interaction between ACE2 and the RBD of all clade 1b and clade 3 sarbecoviruses analyzed.

Claims

1. An antigen-binding molecule, optionally isolated, that binds to a sarbecovirus spike protein, comprising: (i) the following CDRs: HC-CDR1 having the amino acid sequence of SEQ ID NO: 37 HC-CDR2 having the amino acid sequence of SEQ ID NO:53 HC-CDR3 having the amino acid sequence of SEQ ID NO: 54 a heavy chain variable (VH) region incorporating (ii) the following CDRs: LC-CDR1 having the amino acid sequence of SEQ ID NO: 60 LC-CDR2 having the amino acid sequence of SEQ ID NO: 61 LC-CDR3 having the amino acid sequence of SEQ ID NO: 62 A light chain variable (VL) region incorporating An antigen-binding molecule comprising:

2. a VH region having an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 52; and A VL region having an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO:

59. The antigen-binding molecule of claim 1, comprising:

3. The antigen-binding molecule of claim 1 or 2, which is a multispecific antigen-binding molecule and further comprises an antigen-binding domain that binds to an antigen other than a sarbecovirus spike protein.

4. A chimeric antigen receptor (CAR) comprising the antigen-binding molecule of any one of claims 1 to 3.

5. One or more nucleic acids, optionally isolated, encoding the antigen-binding molecule of any one of claims 1 to 3 or the CAR of claim 4.

6. One or more expression vectors comprising one or more nucleic acids of claim 5.

7. A cell comprising the antigen-binding molecule of any one of claims 1 to 3, the CAR of claim 4, one or more nucleic acids of claim 5, or one or more expression vectors of claim 6.

8. A method comprising culturing the cell of claim 7 under conditions suitable for expression of the antigen-binding molecule or CAR by the cell.

9. A composition comprising the antigen-binding molecule of any one of claims 1 to 3, the CAR of claim 4, one or more nucleic acids of claim 5, one or more expression vectors of claim 6, or the cell of claim 7, and a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.

10. The composition of claim 9, further comprising an antigen binding molecule that binds to a sarbecovirus spike protein, and comprises a VH region having an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 824 and a VL region having an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO:

830.

11. A combination comprising: (i) an antigen-binding molecule of any one of claims 1 to 3; and (ii) an antigen-binding molecule that binds to a sarbecovirus spike protein, and comprises a VH region having an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO: 824 and a VL region having an amino acid sequence having at least 70% amino acid sequence identity to SEQ ID NO:

830.

12. 12. The antigen-binding molecule of any one of claims 1 to 3, the CAR of claim 4, the one or more nucleic acids of claim 5, the one or more expression vectors of claim 6, the cell of claim 7, the composition of claim 9 or claim 10, or the combination of claim 11, for use in a method of medical treatment or prophylaxis.

13. The antigen-binding molecule of any one of claims 1 to 3, the CAR of claim 4, the one or more nucleic acids of claim 5, the one or more expression vectors of claim 6, the cell of claim 7, the composition of claim 9 or claim 10, or the combination of claim 11, for use in treating or preventing a disease or condition characterized by infection with a sarbecovirus, wherein the disease or condition characterized by infection with a sarbecovirus may be COVID-19.

14. Use of the antigen-binding molecule of any one of claims 1 to 3, the CAR of claim 4, the one or more nucleic acids of claim 5, the one or more expression vectors of claim 6, the cell of claim 7, the composition of claim 9 or claim 10, or the combination of claim 11 in the manufacture of a medicament for treating or preventing a disease or condition characterized by infection with a sarbecovirus, wherein the disease or condition characterized by infection with a sarbecovirus may be COVID-19.

15. 12. A method for treating or preventing a disease or condition characterized by infection with a sarbecovirus in a subject, comprising the step of administering to the subject a therapeutically or prophylactically effective amount of the antigen-binding molecule of any one of claims 1 to 3, the CAR of claim 4, one or more nucleic acids of claim 5, one or more expression vectors of claim 6, the cell of claim 7, the composition of claim 9 or claim 10, or the combination of claim 11, wherein the disease or condition characterized by infection with a sarbecovirus may be COVID-19.

16. An in vitro complex, optionally isolated, comprising an antigen-binding molecule of any one of claims 1 to 3 bound to a sarbecovirus or a sarbecovirus spike protein.

17. A method for detecting a sarbecovirus or a sarbecovirus spike protein in a sample, the method comprising the steps of contacting a sample containing or suspected of containing a sarbecovirus or a sarbecovirus spike protein with the antigen-binding molecule of any one of claims 1 to 3, and detecting formation of a complex between the antigen-binding molecule and the sarbecovirus or the sarbecovirus spike protein.

18. A method for selecting or stratifying a subject for treatment with a drug that targets a sarbecovirus, the method comprising the steps of contacting a sample from the subject in vitro with the antigen-binding molecule of any one of claims 1 to 3, and detecting formation of a complex between the antigen-binding molecule and the sarbecovirus or a sarbecovirus spike protein.

19. Use of the antigen-binding molecule of any one of claims 1 to 3 as a diagnostic or prognostic agent in vitro or in vivo.

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