Methods for preventing or treating coronavirus infections

Abstract

The present invention is in the field of medical treatment and relates to a method for the treatment of coronavirus infections. In particular, the present invention relates to a method for the prophylactic and / or therapeutic treatment of coronavirus infections by mucosal administration of antibodies against coronaviruses.

Description

[Technical Field]

[0001] The present invention is in the field of medical treatment and relates to methods for the treatment of β-coronavirus infections in animals and humans, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2).

[0002] In particular, the present invention relates to a method for the prophylactic and / or therapeutic treatment of SARS-CoV-2 by transmucosal administration, in particular intranasal and / or oral inhalation, of antibodies against β-coronaviruses, in particular SARS-CoV-2. [Background technology]

[0003] The SARS-CoV-2 virus causes the disease COVID-19 in humans and is spreading rapidly, having a significant impact on human society. The SARS-CoV-2 virus and its variants of concern are expected to continue to cause high rates of infection and severe illness and death. As of January 2023, the ongoing COVID-19 pandemic caused by SARS-CoV-2 has resulted in more than 6.7 million deaths worldwide since 2019.

[0004] In recent years, other coronavirus outbreaks have caused severe illness and death. While scientists are developing therapeutic antibodies and vaccines against SARS-CoV-2, the risk of future emergence of novel SARS-CoV-2 variants and novel coronaviruses remains. These novel coronaviruses could cause morbidity and mortality and could spark pandemics. Identifying broad-spectrum protective therapies that can combat current and future coronavirus outbreaks remains crucial.

[0005] In addition to SARS-CoV-2, four other betacoronaviruses are known to cause disease in humans: betacoronaviruses HCoV-OC43 (human coronavirus OC43), HCoV-HKU1 (human coronavirus HKU1), SARS-CoV (severe acute respiratory syndrome coronavirus), and MERS-CoV (Middle East respiratory syndrome coronavirus). Spillover events, in which humans become infected with coronaviruses circulating in animal reservoirs, are common. Recently, three betacoronaviruses, including SARS-CoV-2, have crossed over from animals to humans, causing severe epidemics.

[0006] Furthermore, two coronaviruses previously associated only with animal infections have recently been detected in humans presenting with influenza-like symptoms.

[0007] The name coronavirus comes from its crown-like appearance. Coronaviruses are a large group of viruses that have spike proteins on their surface that resemble the spikes of a crown. Antibodies elicited by natural infection or by the COVID-19 vaccines used for herd immunity in 2020–2023 primarily target the variable receptor-binding domain (RBD) of these spike proteins.

[0008] Spike protein structure Coronavirus infection is a multistep process involving the enzymatic cleavage and rearrangement of the surface spike protein, which contains an S1 subunit that contains the receptor-binding domain (RBD) and an S2 subunit that is involved in the fusion of viral and (host) cell membranes, thereby facilitating cell entry.

[0009] The SARS-CoV-2 viral spike protein facilitates viral entry primarily by binding to the angiotensin-converting enzyme 2 (ACE2) receptor on host cells.

[0010] The SARS-CoV-2 spike contains two cleavage sites: a furin cleavage site at the interface of the S1 and S2 subunits, and an S2' site that is highly conserved among different coronaviruses.

[0011] S1 subunit The SARS-CoV-2 spike protein uses the RBD on the S1 subunit to engage with the ACE2 receptor on target cells.

[0012] The S1 subunit is more accessible and remains the primary target of many neutralizing antibodies. However, the S1 subunit is more genetically variable than the S2 subunit, especially when subjected to antibody selection pressure. This tendency toward genetic variability can lead to viral variants with major changes occurring in the S1 subunit. Changes in the receptor-binding domain in SARS-CoV-2 variants of concern dramatically reduce protection from antibodies elicited by previous infection and / or vaccines, often resulting in reinfection.

[0013] S2 subunit The viral spike component, which is important for infection, also contains the structurally complex S2 subunit, which contains dynamic elements important for fusion with host cells. Upon receptor binding, the S1 subunit is discarded, and the membrane enzyme transmembrane serine protease 2 (TMPRSS2) or endosomal cathepsins cleave the S2 region.

[0014] This cleavage leads to the insertion of the fusion peptide into the cell membrane, ultimately resulting in viral fusion.

[0015] These S2 subunit elements are less susceptible to genetic variability than the RBD, which has so far been able to retain or even increase its ability to bind ACE2 despite various mutations.

[0016] The S2 domain site presents an inaccessible target for novel therapeutics that protect against a broader range of coronaviruses.

[0017] Stem Helix The stem helix at the base of the viral spike protein is less accessible than the element on the S2 subunit, but has the advantage of being better conserved in amino acid sequence.

[0018] Treatment and Prevention There are limited viral treatment options for SARS-CoV-2, in the form of small molecule or antiviral antibody drugs. Few interventions exist to prevent SARS-CoV-2 infection; most are non-pharmaceutical, such as social distancing and wearing masks. These methods are non-specific, and efficacy is largely dependent on adherence. Therefore, there is a need for different treatments, especially those that simultaneously target multiple SARS-CoV-2 variants of concern, such as broadly neutralizing antibodies (bnAbs).

[0019] Although treatment of SARS-CoV-2 infection with one, two, or more antibodies is feasible, treatments involving a single antibody or a combination of antibodies would be more cost-effective. The currently predominant SARS-CoV-2 omicron subvariant BA.5 is resistant to most monoclonal antibody therapeutics. Betacoronaviruses, including SARS-CoV-2, also infect a range of animal species known to frequently come into contact with humans, thus increasing the risk of animal transmission and novel outbreaks caused by SARS-CoV-2 variants or viruses that have not previously infected humans and for which no treatment exists.

[0020] Therefore, there is an urgent need for the development of therapeutic mAbs that broadly target β-coronaviruses and novel SARS-CoV-2 variants of concern that are not neutralized by antibodies elicited by immunity and / or natural infection. Summary of the Invention [Problem to be solved by the invention]

[0021] Identifying broad-spectrum protective therapies that can combat the novel coronavirus remains crucial, and the inventors have surprisingly developed a novel array of broad-spectrum protective antibodies, as disclosed herein. [Means for solving the problem]

[0022] The present inventors have discovered that antibodies having the complementarity determining regions (CDRs) disclosed herein can be advantageously used in the treatment of coronavirus infections, particularly in the prophylactic treatment of coronavirus infections.

[0023] The inventors have surprisingly established that mucosal administration of antibodies having the CDRs disclosed herein is a more potent route of prophylactic administration compared to intraperitoneal administration when treating coronavirus infections.

[0024] The inventors have surprisingly established that intranasal administration of antibodies having the CDRs disclosed herein is a more potent route of prophylactic administration compared to intraperitoneal administration when treating coronavirus infection.

[0025] Significantly, the transmucosal administration method of the treatment of the present invention is effective against SARS-Cov-2 and variants of concern and does not require the use of additional antibodies.

[0026] The transmucosal treatment method of the present invention is effective at low dosages and prevents weight loss.

[0027] Complementarity-Determining Regions (CDRs) Preferably, the CDR regions are according to Kabat et al. (1991), as described in Sequences of Proteins of Immunological Interest.

[0028] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 257. The present invention provides antibodies comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574.

[0029] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 257; Provided is an antibody comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, or 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, or 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, or 569 to 574, wherein the antibody is administered to a mucosal epithelium.

[0030] In a preferred embodiment, the present invention provides a method for treating a coronavirus infection in an individual, the method comprising administering to an individual a heavy chain variable dopant comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, or 050 to 055, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, or 132 to 137, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, or 252 to 257. The method includes a step of administering to an individual in need thereof an antibody comprising a light chain variable domain including a main, a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363, a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480, and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574, wherein the antibody is administered to the mucosal epithelium.

[0031] In a preferred embodiment, the present invention provides a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 257; Provided is a composition for mucosal application comprising an antibody comprising a light chain variable domain including a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574.

[0032] In a preferred embodiment, the present invention provides an antibody for use in a method for treating a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, or 050 to 055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, or 132 to 137; and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, or 252 to 257. a heavy chain variable domain comprising a heavy chain CDR3 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574, and the antibody is administered to a mucosal epithelium.

[0033] In a preferred embodiment, the invention provides an antibody or a method or composition comprising an antibody, wherein the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising SEQ ID NO: 055, a heavy chain CDR2 region comprising SEQ ID NO: 137, and a heavy chain CDR3 region comprising SEQ ID NO: 257; a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 363, a light chain CDR2 region comprising SEQ ID NO: 480, and a light chain CDR3 region comprising SEQ ID NO: 574.

[0034] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 197-202, 208-213, 219-224, 230-235, 241-246, 252-257. The antibody comprises a heavy chain variable domain including a heavy chain CDR3 region; a light chain CDR1 region including any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region including any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region including any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574, and 0.1 mg to 20 mg of the antibody is administered to the mucosa.

[0035] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, and a heavy chain CDR2 region comprising any one of SEQ ID NOs: 197-202, 208-213, 219-224, 230-235, 241-246, 252-257. The antibody comprises a heavy chain variable domain including a heavy chain CDR3 region; a light chain CDR1 region including any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region including any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region including any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574, and 0.1 mg to 20 mg of the antibody is administered intranasally.

[0036] In a preferred embodiment, disclosed herein is an antibody for use in a method for the prophylactic treatment of a coronavirus infection in an individual, the antibody comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197-202, 208-213, 219-224, 230-235, 241-246, 252-257. The antibody comprises a light chain variable domain comprising a heavy chain variable domain including a DR3 region; a light chain CDR1 region including any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region including any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region including any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574, and 0.1 mg to 20 mg of the antibody is administered by oral inhalation.

[0037] In a preferred embodiment, disclosed herein is a composition comprising an antibody for use in a method for preventing or treating coronavirus infection, wherein the antibody comprises a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, or 050-055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077-082, 088-093, 099-104, 110-115, 121-126, or 132-137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197-202, 208-213, 219-224, 230-235, 241-246, or 252-257. the antibody comprises a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574; the route of administration includes at least one of pulmonary administration, nasal administration, and oropharyngeal administration; and the nominal dose of the antibody is 0.1 mg to 20 mg.

[0038] Preferably, the antibody disclosed herein comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 257.

[0039] Preferably, the antibody disclosed herein comprises a light chain variable domain comprising: a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574.

[0040] Preferably, in the method for treatment of the present invention, the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 257.

[0041] Preferably, in the method for treatment of the present invention, the antibody comprises a light chain variable domain comprising a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574.

[0042] In an alternative preferred embodiment, the present invention provides a heavy chain variable fragment comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 054, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 136, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 256. and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 573.

[0043] Embodiments with more than one CDR Preferably, the antibodies disclosed herein comprise a heavy chain variable domain comprising a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077-082, 088-093, 099-104, 110-115, 121-126, and 132-137.

[0044] Route of administration In a preferred embodiment, the invention provides an antibody disclosed herein or a method or composition comprising an antibody disclosed herein, wherein the antibody is administered by transmucosal administration.

[0045] In a preferred embodiment, the invention provides an antibody disclosed herein, or a method or composition comprising an antibody disclosed herein, wherein the antibody is administered by intravenous administration.

[0046] In a preferred embodiment, the invention provides an antibody disclosed herein or a method or composition comprising an antibody disclosed herein, wherein the antibody is administered by at least one of oral inhalation, nasal administration, intraocular administration, vaginal administration, rectal administration, and oropharyngeal administration.

[0047] In a preferred embodiment, the antibodies disclosed herein are administered intranasally.

[0048] Such antibodies are useful for treating a coronavirus infection in an individual. Preferably, the method for treating a coronavirus infection is a method for prophylactic and / or therapeutic treatment of a coronavirus infection. Preferably, the antibody is provided to an individual infected with a coronavirus. Preferably, the antibody is provided to an individual prophylactically. Preferably, the antibody is provided to an individual after infection but before the onset of symptoms.

[0049] In a preferred embodiment, the present disclosure provides a method of treating a coronavirus infection in an individual, the method comprising administering to an individual a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, or 050-055, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077-082, 088-093, 099-104, 110-115, 121-126, or 132-137, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197-202, 208-213, 219-224, 230-235, 241-246, or 252-257. and a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574, to an individual in need thereof, including transmucosal administration, preferably intranasal administration and / or oral inhalation.

[0050] In a preferred embodiment, the individual is infected with or at risk for coronavirus infection.

[0051] In a preferred embodiment, the present disclosure provides an antibody disclosed herein for use in the manufacture of a medicament for use in the treatment of a coronavirus infection.

[0052] section Each of the following numbered sections represents a preferred embodiment of the present invention and is part of this specification. Verse 1: 1. A method for treating a coronavirus infection in an individual, comprising administering to an individual a heavy chain variable domain comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, or 050-055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077-082, 088-093, 099-104, 110-115, 121-126, or 132-137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197-202, 208-213, 219-224, 230-235, 241-246, or 252-257; a light chain CDR1 region comprising any one of SEQ ID NOs: 3 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574, wherein the antibody is administered mucosally to an individual in need thereof. Verse 2: a heavy chain variable domain comprising a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 055, a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 137, and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 257; A composition for mucosal application comprising an antibody comprising a light chain variable domain comprising a light chain CDR1 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480, and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. Verse 3: 1. An antibody for use in a method for preventing or treating a coronavirus infection in an individual, comprising: a heavy chain CDR1 region comprising any one of SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, or 050 to 055; a heavy chain CDR2 region comprising any one of SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, or 132 to 137; and a heavy chain CDR3 region comprising any one of SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, or 252 to 257. an antibody for mucosal administration, the antibody comprising a light chain variable domain including a heavy chain variable domain containing an R3 region; a light chain CDR1 region comprising any one of SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363; a light chain CDR2 region comprising any one of SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480; and a light chain CDR3 region comprising any one of SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. Verse 4: The method, composition, or antibody according to any preceding clause, wherein the antibody comprises a heavy chain variable domain comprising a heavy chain CDR1 region comprising SEQ ID NO: 055, a heavy chain CDR2 region comprising SEQ ID NO: 137, and a heavy chain CDR3 region comprising SEQ ID NO: 257; a light chain variable domain comprising a light chain CDR1 region comprising SEQ ID NO: 363, a light chain CDR2 region comprising SEQ ID NO: 480, and a light chain CDR3 region comprising SEQ ID NO: 574. Verse 5: The method, composition, or antibody according to any preceding clause, wherein the method for treatment of coronavirus virus infection is a method for prophylactic and / or therapeutic treatment of coronavirus, preferably the method for treatment is for prophylactic treatment of SARS-COV-2 infection. Verse 6: The method, composition, or antibody according to any preceding clause, wherein the antibody comprises a heavy chain variable domain having the sequence of SEQ ID NO: 629 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, amino acid insertions, deletions, or substitutions that are not in the heavy chain CDRs. Verse 7: The method, composition, or antibody according to any preceding clause, wherein the antibody comprises a light chain variable domain having the sequence of SEQ ID NO: 630 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, amino acid insertions, deletions, or substitutions that are not in the light chain CDRs. Verse 8: The method, composition, or antibody according to any preceding clause, wherein the antibody is an IgG antibody, preferably an IgG1 antibody. Verse 9: The method, composition, or antibody according to any preceding clause, wherein the antibody is provided prophylactically to the individual. Verse 10: The method, composition, or antibody according to any preceding clause, wherein the route of administration comprises at least one of oral inhalation, nasal administration, intraocular administration, and oropharyngeal administration. Verse 11: The method, composition, or antibody according to any preceding clause, wherein the antibody is administered at least once or at least twice monthly. Verse 12: The method, composition, or antibody according to any preceding clause, wherein the antibody is administered to the individual in a dosage of between 0.01 mg and 20 mg. Verse 13: 16. A composition according to any preceding clause comprising a single dose unit of 0.01 mg to 20 mg, preferably 0.1 mg to 15 mg, or preferably 0.5 mg to 10 mg of the antibody, wherein the antibody is defined in any one of clauses 1 to 12. Verse 14: A composition according to any preceding clause that is self-administered. Verse 15: A pharmaceutical delivery device comprising a composition according to any preceding clause.

[0053] Dosage Preferably, 0.01 mg to 20 mg of antibody is administered to an individual, and more preferably, 0.1 mg to 20 mg of antibody is administered to an individual.

[0054] The fixed dose, or nominal dose, of the antibody is preferably 0.01 mg to 17 mg, preferably 0.011 mg to 16 mg, preferably 0.012 mg to 15 mg, preferably 0.013 mg to 14 mg, preferably 0.014 mg to 13 mg, preferably 0.015 mg to 12 mg, preferably 0.016 mg to 11 mg, preferably 0.017 mg to 10 mg, preferably 0.018 mg to 9 mg, preferably 0.020 mg to 8 mg, preferably 0.023 mg to 7 mg, preferably 0.025 mg to 6 mg, preferably 0.030 mg to 5 mg, preferably 0.040 mg to 4 mg, preferably 0.050 mg to 3 mg, preferably 0.075 mg to 2 mg, or preferably 0.10 mg to 1 mg.

[0055] The fixed dose, or nominal dose, of the antibody is preferably 0.010 mg to 5.0 mg, preferably 0.020 mg to 4.5 mg, preferably 0.030 mg to 4.0 mg, preferably 0.040 mg to 3.5 mg, preferably 0.050 mg to 3.0 mg, preferably 0.060 mg to 2.5 mg, preferably 0.070 mg to 2.0 mg, preferably 0.080 mg to 1.5 mg, preferably 0.090 mg to 1.0 mg, or preferably 0.100 mg to 0.5 mg.

[0056] The fixed dose, or nominal dose, of the antibody is preferably 0.010 mg to 15.0 mg, preferably 0.020 mg to 14.5 mg, preferably 0.030 mg to 4.0 mg, preferably 0.040 mg to 13.5 mg, preferably 0.050 mg to 13.0 mg, preferably 0.060 mg to 12.5 mg, preferably 0.070 mg to 12.0 mg, preferably 0.080 mg to 11.5 mg, preferably 0.090 mg to 11.0 mg, or preferably 0.100 mg to 10.5 mg.

[0057] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 0.1 μg to 10 μg, preferably 0.2 μg to 9 μg, preferably 0.3 μg to 8 μg, preferably 0.4 μg to 7 μg, preferably 0.5 μg to 6 μg, preferably 0.6 μg to 5 μg, preferably 0.7 μg to 4 μg, preferably 0.8 μg to 3 μg, preferably 0.9 μg to 2 μg, or preferably about 1 μg.

[0058] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 1 mg to 11 mg, preferably 2 mg to 12 mg, preferably 3 mg to 13 mg, preferably 4 mg to 14 mg, preferably 5 mg to 15 mg, preferably 6 mg to 16 mg, preferably 7 mg to 17 mg, preferably 8 mg to 18 mg, preferably 9 mg to 19 mg, or preferably 10 mg to 20 mg.

[0059] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 1 mg to 20 mg, preferably 2 mg to 19 mg, preferably 3 mg to 18 mg, preferably 4 mg to 17 mg, preferably 5 mg to 16 mg, preferably 6 mg to 15 mg, preferably 7 mg to 14 mg, preferably 8 mg to 13 mg, preferably 9 mg to 12 mg, or preferably 10 mg to 11 mg.

[0060] The fixed dose, or nominal dose, of antibody to be delivered to a human subject is preferably 11 mg to 20 mg, preferably 10 mg to 19 mg, preferably 9 mg to 18 mg, preferably 8 mg to 17 mg, preferably 7 mg to 16 mg, preferably 6 mg to 15 mg, preferably 5 mg to 14 mg, preferably 4 mg to 13 mg, preferably 3 mg to 12 mg, preferably 2 mg to 11 mg, or preferably 1 mg to 10 mg.

[0061] In preferred embodiments, the present disclosure further provides compositions formulated for mucosal administration comprising an antibody disclosed herein in a single dosage unit of between 0.1 mg and 20 mg, preferably between 5 mg and 15 mg, or preferably between 7.5 mg and 12.5 mg.

[0062] In preferred embodiments, the present disclosure further provides compositions formulated for intranasal administration comprising an antibody disclosed herein in a single dosage unit of between 0.1 mg and 20 mg, preferably between 5 mg and 15 mg, or preferably between 7.5 mg and 12.5 mg.

[0063] In preferred embodiments, the present disclosure further provides compositions formulated for oral inhalation comprising an antibody disclosed herein in a single dosage unit of between 0.1 mg and 20 mg, preferably between 5 mg and 15 mg, or preferably between 7.5 mg and 12.5 mg.

[0064] Timing and Spacing Preferably, the antibody is administered at least once or at least twice per month.

[0065] Preferably, the antibody is administered at least once or at least twice per week.

[0066] The fixed dose, or nominal dose, of antibody per week is 0.010 mg to 17 mg, preferably 0.011 mg to 16 mg, preferably 0.012 mg to 15 mg, preferably 0.013 mg to 14 mg, preferably 0.014 mg to 13 mg, preferably 0.015 mg to 12 mg, preferably 0.016 mg to 11 mg, preferably 0.017 mg to 10 mg, preferably 0.018 mg to 9 mg, preferably 0.020 mg to 8 mg, preferably 0.023 mg to 7 mg, preferably 0.025 mg to 6 mg, preferably 0.030 mg to 5 mg, preferably 0.040 mg to 4 mg, preferably 0.050 mg to 3 mg, preferably 0.075 mg to 2 mg, or preferably 0.10 mg to 1 mg.

[0067] The fixed dose, or nominal dose, of antibody per week is 0.010 mg to 5.0 mg, preferably 0.020 mg to 4.5 mg, preferably 0.030 mg to 4.0 mg, preferably 0.040 mg to 3.5 mg, preferably 0.050 mg to 3.0 mg, preferably 0.060 mg to 2.5 mg, preferably 0.070 mg to 2.0 mg, preferably 0.080 mg to 1.5 mg, preferably 0.090 mg to 1.0 mg, or preferably 0.100 mg to 0.5 mg.

[0068] Preferably, the antibody is administered at least once or at least twice per day. Preferably, the antibody is administered daily. Preferably, the antibody is administered twice daily.

[0069] Preferably, the daily fixed dose, or nominal dose, is between 0.001 mg and 17 mg, preferably between 0.011 mg and 16 mg, preferably between 0.012 mg and 15 mg, preferably between 0.013 mg and 14 mg, preferably between 0.014 mg and 13 mg, preferably between 0.015 mg and 12 mg, preferably between 0.016 mg and 11 mg, preferably between 0.017 mg and 10 mg, preferably between 0.018 mg and 9 mg, preferably between 0.020 mg and 8 mg, preferably between 0.023 mg and 7 mg, preferably between 0.025 mg and 6 mg, preferably between 0.030 mg and 5 mg, preferably between 0.040 mg and 4 mg, preferably between 0.050 mg and 3 mg, preferably between 0.075 mg and 2 mg, or preferably between 0.10 mg and 1 mg.

[0070] Framework Area Preferably, the antibody comprises a heavy chain variable domain having the sequence of SEQ ID NO: 629 and / or a light chain variable domain having the sequence of SEQ ID NO: 630.

[0071] Preferably, the heavy chain variable domain of the antibody further comprises a heavy chain framework region FR1 of SEQ ID NO: 631, a heavy chain framework region FR2 of SEQ ID NO: 632, a heavy chain framework region FR3 of SEQ ID NO: 633, and / or a heavy chain framework region FR4 of SEQ ID NO: 634.

[0072] Preferably, the light chain variable domain further comprises a light chain framework region RF1 of SEQ ID NO: 635, a light chain framework region FR2 of SEQ ID NO: 636, a light chain framework region FR3 of SEQ ID NO: 637, and / or a light chain framework region FR4 of SEQ ID NO: 638.

[0073] Combination Compositions In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-220, 221-222, 222-224, 223-225, 224-226, 225-228, 226-228, 227-229, 230-231, 232-233, 234-235, 236-237, 238-239, 240-242, 242-243, 244-245, 246-247, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 273-274, 275-276, 277-278, 279-280, 281-282, 282-283, 283-284, 285-286, 2 24, 230 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, 569 to 574. More preferably, the first fragment comprises a sequence comprising one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 639, 640, 641, 642, 643, or SEQ ID NO: 644. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0074] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-220, 221-222, 222-224, 223-225, 224-226, 225-228, 226-228, 227-229, 230-231, 232-233, 234-235, 236-237, 238-239, 240-242, 242-243, 244-245, 246-247, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 273-274, 275-276, 277-278, 279-280, 281-282, 282-283, 283-284, 285-286, 2 24, 230 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, 569 to 574. More preferably, the first fragment comprises one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 645, 646, 647, 648, 649, or SEQ ID NO: 650. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0075] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-220, 221-222, 222-224, 223-225, 224-226, 225-228, 226-228, 227-229, 230-231, 232-233, 234-235, 236-237, 238-239, 240-242, 242-243, 244-245, 246-247, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 273-274, 275-276, 277-278, 279-280, 281-282, 282-283, 283-284, 285-286, 2 24, 230 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, 569 to 574. More preferably, the first fragment comprises a sequence comprising one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 651, 652, 653, 654, 655, or SEQ ID NO: 656. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0076] In a preferred embodiment, the inventors disclose a composition comprising a first binding fragment disclosed herein and a second binding fragment, wherein the first fragment is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-220, 221-222, 222-224, 223-225, 224-226, 225-228, 226-228, 227-229, 230-231, 232-233, 234-235, 236-237, 238-239, 240-242, 242-243, 244-245, 246-247, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 273-274, 275-276, 277-278, 279-280, 281-282, 282-283, 283-284, 285-286, 2 24, 230 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first fragment comprises a sequence comprising one or more of SEQ ID NOs: 055, 137, 257, 363, 480, and 574. The second binding fragment comprises a sequence comprising any one of SEQ ID NOs: 657, 658, 659, 660, 661, or SEQ ID NO: 662. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0077] bispecific antibody In a preferred embodiment, the inventors disclose a bispecific antibody, which is capable of binding to the stem helix of a coronavirus. In a preferred embodiment, the inventors disclose a bispecific antibody, which is capable of binding to the stem helix of SARS-CoV-2.

[0078] In a preferred embodiment, the inventors disclose an anti-coronavirus bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of a coronavirus and a second Fab capable of binding to the fusion peptide of a coronavirus. In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2.

[0079] In a preferred embodiment, we disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of the preferred SEQ ID NOs: 055, 137, 257, 363, 480, or 574, or the first Fab comprises a sequence comprising any one or more of the preferred SEQ ID NOs: 651, 652, 653, 654, 655, 656, 657, 658, 659, 660, 661, 662, 663, 664, 665, 666, 667, 668, 669, 670, 671, 672, 673, 674, 675, 676, 677, 678, 679, 680, 681, 682, 683, 684, 685, 686, 687, 688, 689, 690, 700, 701, 702, 703, 704, 705, 706, 707, 708, 710, 711, 712, 713, 714, 715, 716, 717, 718, 719, 720, 721, 722, 723, 724, 725, 726, 727, 728, 729, 730, 731, 732, 733, Alternatively, the first Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 657, 658, 659, 660, 661-662, and the second Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 639, 640, 641, 642, 643-644, or the second Fab comprises a sequence comprising one or more of preferred SEQ ID NOs: 645, 646, 647, 648, 649-650.

[0080] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 639, 640, 641, 642, 643-644. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0081] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 645, 646, 647, 648, 649-650. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0082] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab also capable of binding to the stem helix of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 651, 652, 653, 654, 655-656. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0083] In a preferred embodiment, the inventors disclose an anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab also capable of binding to the stem helix of SARS-CoV-2, wherein the first Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second Fab comprises a sequence comprising any one or more of preferred SEQ ID NOs: 657, 658, 659, 660, 661-662. Preferably, the fragment is selected from any one of the group comprising a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab'), an Fv, a single-domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

[0084] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 055, 137 or 257, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 660, 661 or 662.

[0085] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 055, 137 or 257, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 642, 643 or 644.

[0086] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 055, 137 or 257, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 648, 649 or 650.

[0087] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 055, 137 or 257, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 654, 655 or 656.

[0088] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 657, 658 or 659, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 363, 480 or 574.

[0089] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 639, 640 or 641, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 363, 480 or 574.

[0090] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 645, 646 or 647, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 363, 480 or 574.

[0091] In a preferred embodiment, we disclose a bispecific antibody comprising a first Fab and a second Fab, wherein the first Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 651, 652 or 653, and the second Fab comprises any one or more of the sequences set forth in preferred SEQ ID NOs: 363, 480 or 574.

[0092] The present invention also provides methods for the construction, expression, and purification of bispecific antibodies having the function of binding to the stem helix of SARS-CoV-2, and the use of bispecific antibodies in the field of medicine, particularly in the prevention and / or treatment of SARS-CoV-2 infection.

[0093] section Each of the following numbered sections represents a preferred embodiment of the present invention and is part of this specification. Verse 26: An anti-coronavirus, preferably anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 639, 640, 641, 642, 643-644. Verse 27: 27. The bispecific antibody or antigen-binding fragment thereof according to clause 26, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, and / or 574, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, and / or 644. Verse 28: a first Fab that specifically binds to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2; a.) a first Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 055, the heavy chain CDR2 region of SEQ ID NO: 137, and the heavy chain CDR3 region of SEQ ID NO: 257, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 363, the light chain CDR2 region of SEQ ID NO: 480, and the light chain CDR3 region of SEQ ID NO: 574; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644; 28. The bispecific antibody or antigen-binding fragment thereof according to clause 26 or 27, Verse 29: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2; and b) a light chain comprising the VL-CL domains and a heavy chain comprising the VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2; Including, the constant domains CL and CH1 from an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 29. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 28. Verse 30: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 055, a heavy chain CDR2 region of SEQ ID NO: 137, and a heavy chain CDR3 region of SEQ ID NO: 257; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 363, a light chain CDR2 region of SEQ ID NO: 480, and a light chain CDR3 region of SEQ ID NO: 574; The VH domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644. 29. A bispecific antibody or antigen-binding fragment thereof according to clause 29. Verse 31: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the constant domains CL and CH1 from an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 29. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 28. Verse 32: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 055, a heavy chain CDR2 region of SEQ ID NO: 137, and a heavy chain CDR3 region of SEQ ID NO: 257; and the VL domain of the antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 363, a light chain CDR2 region of SEQ ID NO: 480, and a light chain CDR3 region of SEQ ID NO: 574; The VH domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641, and the VL domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644; 32. A bispecific antibody or antigen-binding fragment thereof according to clause 31. Verse 33: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the domains VL-CL and VH-CH1 from an antibody that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 29. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 28. Verse 34: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the first Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 629. Verse 35: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the second Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 673. Verse 36: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the first Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 630. Verse 37: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, wherein the second Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO:674. Verse 38: 34. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 33, selected from any one of the group comprising a full length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof. Verse 39: 39. A method for the production of a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, comprising: a) culturing a host cell comprising an expression vector comprising a polynucleotide encoding the bispecific antibody or antibody fragment according to any one of the preceding clauses under conditions that allow the production of said bispecific antibody or antigen-binding fragment thereof, and b) isolating the bispecific antibody or antigen-binding fragment thereof. A method comprising: Verse 40: 39. A nucleic acid molecule comprising a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, which is preferably an isolated nucleic acid molecule. Verse 41: a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38 and clause 40, comprising a nucleic acid sequence encoding the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof, The nucleic acid sequence is (i) a first nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 055, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 055 by at most one or two amino acids; (ii) a second nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 137, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 137 by at most 1, 2, 3, 4, or 5 amino acids; (iii) a third nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 257, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 257 by at most 1, 2, or 3 amino acids; (iv) a fourth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 363, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 363 by at most 1, 2, 3, or 4 amino acids; (v) a fifth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 480, or an amino acid sequence that differs from SEQ ID NO: 480 by at most one or two amino acids; (vi) a sixth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 574, or an amino acid sequence that differs from SEQ ID NO: 574 by at most 1, 2, 3, or 4 amino acids; (vii) a seventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 639, or an amino acid sequence that differs from SEQ ID NO: 639 by at most one or two amino acids; (viii) an eighth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 640, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 640 by at most 1, 2, 3, 4, or 5 amino acids; (ix) a ninth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 641, or an amino acid sequence that differs from SEQ ID NO: 641 by at most 1, 2, 3, or 4 amino acids; (x) a tenth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 642, or an amino acid sequence that differs from SEQ ID NO: 642 by at most 1, 2, 3, or 4 amino acids; (xi) an eleventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 643, or an eleventh nucleic acid sequence encoding an amino acid sequence differing from SEQ ID NO: 643 by at most one or two amino acids; and (xii) a twelfth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 644, or an amino acid sequence that differs from SEQ ID NO: 644 by at most 1, 2, or 3 amino acids; A nucleic acid molecule comprising at least one of the following: Verse 42: 39. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38 for use in a method of treating a coronavirus infection in a subject. Verse 43: 39. A composition comprising a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38. Verse 44: 44. A composition according to clause 43 further comprising a second medicament for simultaneous, separate or sequential administration. Verse 45: 45. The composition according to clause 44, wherein the second medicament comprises a second antibody or antigen-binding fragment thereof. Verse 46: 45. The composition according to clause 44, wherein the second medicament comprises a bispecific antibody or a bispecific antigen-binding fragment thereof. Verse 47: 44. The composition according to clause 43, further comprising a pharmaceutically acceptable excipient or carrier. Verse 48: 44. A composition according to clause 43 for use in preventing and / or treating a coronavirus infection in a subject, preferably a SARS-CoV-1 or SARS-CoV-2 infection in a subject, comprising an effective amount of the bispecific antibody or antigen-binding fragment thereof, nucleic acid molecule, or vector, and optionally a pharmaceutically acceptable excipient or carrier. Verse 49: 44. The composition of clause 43 for use as a medicament. Verse 50: 44. An inhalation device comprising the composition of clause 43. Verse 51: 39. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, which is bivalent. Verse 52: 39. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 26 to 38, capable of binding to at least one stem helix and / or fusion peptide of an α-coronavirus, a β-coronavirus, a γ-coronavirus, and a δ-coronavirus, preferably at least an α-coronavirus and / or a β-coronavirus.

[0094] section Each of the following numbered sections represents a preferred embodiment of the present invention and is part of this specification. Verse 53: An anti-coronavirus, preferably anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 645, 646, 647, 648, 649 to 650. Verse 54: 54. The bispecific antibody or antigen-binding fragment thereof according to clause 53, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, and / or 574, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 645, 646, 647, 648, 649, and / or 650. Verse 55: a first Fab that specifically binds to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2; a.) a first Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 055, the heavy chain CDR2 region of SEQ ID NO: 137, and the heavy chain CDR3 region of SEQ ID NO: 257, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 363, the light chain CDR2 region of SEQ ID NO: 480, and the light chain CDR3 region of SEQ ID NO: 574; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 645, the heavy chain CDR2 region of SEQ ID NO: 646, and the heavy chain CDR3 region of SEQ ID NO: 647, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 648, the light chain CDR2 region of SEQ ID NO: 649, and the light chain CDR3 region of SEQ ID NO: 650; 55. The bispecific antibody or antigen-binding fragment thereof according to clause 53 or 54, Verse 56: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2; and b) a light chain comprising the VL-CL domains and a heavy chain comprising the VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2; Including, 56. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 55, wherein the constant domains CL and CH1 from an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, are replaced by each other. Verse 57: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 055, a heavy chain CDR2 region of SEQ ID NO: 137, and a heavy chain CDR3 region of SEQ ID NO: 257; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 363, a light chain CDR2 region of SEQ ID NO: 480, and a light chain CDR3 region of SEQ ID NO: 574; The VH domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 645, the heavy chain CDR2 region of SEQ ID NO: 646, and the heavy chain CDR3 region of SEQ ID NO: 647, and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the light chain CDR1 region of SEQ ID NO: 648, the light chain CDR2 region of SEQ ID NO: 649, and the light chain CDR3 region of SEQ ID NO: 650. 57. A bispecific antibody or antigen-binding fragment thereof according to clause 56. Verse 58: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the constant domains CL and CH1 from an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 56. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 55. Verse 59: the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 055, a heavy chain CDR2 region of SEQ ID NO: 137, and a heavy chain CDR3 region of SEQ ID NO: 257; and the VL domain of the antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 363, a light chain CDR2 region of SEQ ID NO: 480, and a light chain CDR3 region of SEQ ID NO: 574; The VH domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the heavy chain CDR1 region of SEQ ID NO: 645, the heavy chain CDR2 region of SEQ ID NO: 646, and the heavy chain CDR3 region of SEQ ID NO: 647, and the VL domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the light chain CDR1 region of SEQ ID NO: 648, the light chain CDR2 region of SEQ ID NO: 649, and the light chain CDR3 region of SEQ ID NO: 650. 59. A bispecific antibody or antigen-binding fragment thereof according to clause 58. Verse 60: a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the domains VL-CL and VH-CH1 from an antibody that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 56. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 55. Verse 61: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the first Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 629. Verse 62: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the second Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 675. Verse 63: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the first Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 630. Verse 64: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, wherein the second Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO: 676. Verse 65: 61. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 60, selected from any one of the group comprising a full length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof. Verse 66: 66. A method for the production of a bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, comprising: a) culturing a host cell comprising an expression vector comprising a polynucleotide encoding the bispecific antibody or antibody fragment according to any one of the preceding clauses under conditions that allow the production of said bispecific antibody or antigen-binding fragment thereof, and b) isolating the bispecific antibody or antigen-binding fragment thereof. A method comprising: Verse 67: 66. A nucleic acid molecule comprising a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, which is preferably an isolated nucleic acid molecule. Verse 68: 68. A bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65 and clause 67, a nucleic acid molecule comprising a nucleic acid sequence encoding the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof, The nucleic acid sequence is (i) a first nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 055, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 055 by at most one or two amino acids; (ii) a second nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 137, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 137 by at most 1, 2, 3, 4, or 5 amino acids; (iii) a third nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 257, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 257 by at most 1, 2, or 3 amino acids; (iv) a fourth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 363, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 363 by at most 1, 2, 3, or 4 amino acids; (v) a fifth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 480, or an amino acid sequence that differs from SEQ ID NO: 480 by at most one or two amino acids; (vi) a sixth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 574, or an amino acid sequence that differs from SEQ ID NO: 574 by at most 1, 2, 3, or 4 amino acids; (vii) a seventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 645, or an amino acid sequence that differs from SEQ ID NO: 645 by at most one or two amino acids; (viii) an eighth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 646, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 646 by at most 1, 2, 3, 4, or 5 amino acids; (ix) a ninth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 647, or an amino acid sequence that differs from SEQ ID NO: 647 by at most 1, 2, 3, 4, or 5 amino acids; (x) a tenth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 648, or an amino acid sequence that differs from SEQ ID NO: 648 by at most 1, 2, or 3 amino acids; (xi) an eleventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 649, or an eleventh nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 649 by at most one or two amino acids; and (xii) a twelfth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 650, or an amino acid sequence that differs from SEQ ID NO: 650 by at most 1, 2, or 3 amino acids. A nucleic acid molecule comprising at least one of the following: Verse 69: 66. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65 for use in a method of treating a coronavirus infection in a subject. Verse 70: 66. A composition comprising a bispecific antibody and / or antigen-binding fragment thereof according to any one of clauses 53 to 65. Verse 71: 71. A composition according to paragraph 70 further comprising a second medicament for simultaneous, separate or sequential administration. Verse 72: 72. The composition according to clause 71, wherein the second medicament comprises a second antibody or antigen-binding fragment thereof. Verse 73: 72. The composition according to clause 71, wherein the second medicament comprises a bispecific antibody or a bispecific antigen-binding fragment thereof. Verse 74: 71. The composition according to clause 70, further comprising a pharmaceutically acceptable excipient or carrier. Verse 75: 71. A composition according to clause 70 for use in preventing and / or treating a coronavirus infection in a subject, preferably a SARS-CoV-1 or SARS-CoV-2 infection in a subject, comprising an effective amount of the bispecific antibody or antigen-binding fragment thereof, nucleic acid molecule, or vector, and optionally a pharmaceutically acceptable excipient or carrier. Verse 76: 71. The composition of clause 70 for use as a medicament. Verse 77: An inhalation device comprising the composition of clause 70. Verse 78: 66. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, which is bivalent. Verse 79: 66. The bispecific antibody or antigen-binding fragment thereof according to any one of clauses 53 to 65, capable of binding to at least one stem helix and / or fusion peptide of an α-coronavirus, a β-coronavirus, a γ-coronavirus, and a δ-coronavirus, preferably at least an α-coronavirus and / or a β-coronavirus. [Brief explanation of the drawings]

[0095] [Figure 1] Survival after lethal challenge - Kaplan-Meier survival curve for CV3-25 intraperitoneal prophylactic treatment group. Animals (n=10 per group) were treated at dose-adjusted intervals with the control antibody CV3-25 on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. The lines are slightly shifted on the y-axis to improve visual representation. [Figure 2]Survival after lethal challenge - Kaplan-Meier survival curves for CV3-25 intranasal prophylactic treatment groups. Animals (n=10 per group, except for n=8 at 15 mg / kg) were treated at dose titration with control antibody CV3-25 on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. Lines are slightly shifted on the y-axis to improve visual representation. [Figure 3] Survival after lethal challenge - Kaplan-Meier survival curves for the antibody according to the invention intraperitoneal prophylactic treatment group. Animals (n=10 per group) were treated at dose titration with an antibody having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574 on day 1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. The lines are slightly offset on the Y-axis to improve visual representation. [Figure 4A] Survival after lethal challenge - antibodies according to the invention Survival and body weight change for intraperitoneal administration of antibodies according to the invention comprising the following CDRs set forth in SEQ ID NOs: 055, 137, 257, 363, 480, and 574 in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=10 per treatment group) were treated intraperitoneally with titrated antibodies according to the invention on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS, n=10) was included. Kaplan-Meier survival curves of treatment groups are shown. Lines have been slightly shifted on the Y-axis to improve visual presentation. [Figure 4B]Survival after lethal challenge - antibodies according to the invention Survival and body weight change for intraperitoneal administration of antibodies according to the invention comprising the following CDRs set forth in SEQ ID NOs: 055, 137, 257, 363, 480, and 574 in a pre-exposure efficacy study in K18-hACE2 mice challenged with a lethal SARS-CoV-2 delta. Animals (n=10 per treatment group) were treated intraperitoneally with titrated antibodies of the invention on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS, n=10) was included. Body weight change (%) relative to day 0 is shown. Error bars represent the mean and 95% confidence interval. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. Asterisks indicate significant differences compared to the control group. [Figure 5] Survival after lethal challenge - Kaplan-Meier survival curves for the antibody according to the invention intranasal prophylactic treatment group. Animals (n=10 per group) were treated at dose titration with an antibody having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574 on day 1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. The lines are slightly offset on the Y-axis to improve visual representation. [Figure 6A] Figure 1 shows survival and body weight change following intranasal administration of antibodies according to the invention comprising the following CDRs set forth in SEQ ID NOs: 055, 137, 257, 363, 480, and 574 in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=10 per treatment group, except for n=9 at the 0.6 mg / kg dose) were treated intranasally with titrated antibodies according to the invention on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS, n=10) was included. Kaplan-Meier survival curves of the treatment groups are shown. Lines have been slightly shifted on the Y-axis to improve visual presentation. [Figure 6B] Survival and body weight change following intranasal administration of antibodies according to the invention comprising the following CDRs set forth in SEQ ID NOs: 055, 137, 257, 363, 480, and 574 in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=10 per treatment group, except for n=9 at the 0.6 mg / kg dose) were treated intranasally with titrated antibodies according to the invention on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS, n=10) was included. % body weight change relative to day 0 is shown. Error bars represent the mean and 95% confidence interval. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 7] Survival and body weight change following intranasal administration of two parental antibodies and a bispecific antibody according to the invention. Survival and body weight change following intranasal administration of two parental antibodies and a bispecific antibody according to the invention in a pre-exposure efficacy study in K18-hACE2 mice challenged with lethal SARS-CoV-2 delta. Animals (n=8 per group) were treated intranasally on day -1 with titrated parental mAb 1 (Figures 7A-7C), parental mAb 2 (Figures 7D-7F), or a bispecific antibody (Figures 7G-7I). Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS, n=10) was included. Figures 7A, 7D, and 7G: Kaplan-Meier survival curves for treatment groups are shown. Lines are slightly shifted relative to the Y-axis for improved visual presentation. Figures 7B, 7E, and 7H: Weight change (%) relative to day 0 are shown. Error bars represent the mean and 95% confidence interval. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. Figures 7G, ​​7H, and 7I: Survival dose-response curves using Probit regression without assuming parallel dose-response curves between treatments. A vehicle control group was included in the modeling at a dose of 0 mg / kg. Pointwise 95% reference confidence intervals (CI) for the predicted doses are shown. Asterisks indicate significant differences compared to the control group. [Figure 8] Body weight change - % body weight change relative to day 0 for the CV3-25 intraperitoneal prophylactic treatment group. Animals (n=10 per group) were treated intranasally with titrated doses of the control antibody CV3-25 on day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. Bars represent the 95% CI of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 9] Body Weight Change - % Body Weight Change relative to Day 0 for the CV3-25 intranasal prophylactic treatment groups. Animals (n=10 per group, except for n=8 in the 15 mg / kg group) were treated intranasally with titrated doses of the control antibody CV3-25 on Day -1. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on Day 0. A vehicle control group (PBS) was included. Bars represent the 95% CI of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 10] Body Weight Change - Antibody According to the Invention Body weight change (%) relative to day 0 for the intraperitoneal prophylactic treatment group. On day -1, animals (n=10 per group) were treated intranasally with a titrated antibody having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. Bars represent the 95% CI of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 11]Body Weight Change - Antibody According to the Invention Body weight change (%) relative to day 0 for the intranasal prophylactic treatment group. On day -1, animals (n=10 per group) were treated intranasally with a titrated antibody having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574. Animals were infected with 103.5 TCID50 of SARS-CoV-2 delta on day 0. A vehicle control group (PBS) was included. Bars represent the 95% CI of the mean. If mice died or were euthanized during study follow-up, the last observed body weight was carried forward. [Figure 12] Linear epitope mapping to the spike helix domains of alpha- and beta-coronaviruses. Amino acids highlighted in gray were identified as epitopes for antibody binding to the spike protein. Target sequences for the seven strains tested were aligned by Clustal. Asterisks indicate that the sequence may contain multiple small portions of the epitope. Epitope footprint (Figure 12A) and paratope hotspots (Figure 12B) for antibodies against the SH domain of SARS-CoV-2. CDRs for the heavy and light chains are shown. [Figure 13A] Graphical representation of the data presented in Example 17. Graphical representation of the extent to which parental mAb 1 (filled triangles) and parental mAb 2 (open triangles), bispecific antibody Bispecific 1 according to the invention (filled circles), bispecific antibody Bispecific 2 according to the invention (open diamonds), bispecific antibody Bispecific 3 according to the invention (open squares), and anti-S1 mAb DZIF-10c (x) bind to selected alpha- and beta-coronaviruses. [Figure 13B] Graphical representation of data presented in Example 17. Graphical representation of the extent to which parental mAb 1 (closed triangles) and parental mAb 2 (open triangles), and bispecific antibody Bispecific 1 (closed circles) bind to selected omicron variants. [Figure 13C]Graphical representation of the data presented in Example 17. Graphical representation of the extent to which the parental mAb 1 (filled triangles), the bispecific antibody Bispecific 1 according to the invention (filled circles), the bispecific antibody Bispecific 2 according to the invention (open diamonds), and the bispecific antibody Bispecific 3 according to the invention (open squares) bind to selected omicron variants. DETAILED DESCRIPTION OF THE INVENTION

[0096] For purposes of clarity and brevity, features may be described herein as part of the same or separate embodiments; however, it will be recognized that the present disclosure includes embodiments having all or any combination of the described features. References to "methods for treatment," "method of treatment," "antibody for use," and "use of an antibody in the manufacture of a medicament" may be used interchangeably, and embodiments disclosed with respect to any one of these aspects also apply with respect to the other aspects. In other words, they all refer to medical treatments involving the antibodies disclosed herein.

[0097] Preventive measures As used herein, the term "prophylactic treatment" includes reference to treatment to prevent an individual from contracting a coronavirus, or to prevent symptoms after a coronavirus infection, or to prevent severe symptoms after a known coronavirus infection, with or without symptoms, or to prevent hospitalization and death after a coronavirus infection.

[0098] Prevention of infection is preferably achieved by administration of the antibodies disclosed herein prior to exposure to coronavirus, i.e., pre-exposure prophylaxis.

[0099] Thus, "prophylactically" preferably means prior to viral exposure, although it can also include administration after infection, e.g., to reduce viral replication or spread or increase viral clearance, i.e., post-exposure prophylaxis.

[0100] An infected individual may be asymptomatic, or alternatively, an infected individual may be symptomatic.

[0101] Preferably, post-exposure prophylaxis involves administration of the antibodies disclosed herein after exposure to the coronavirus to prevent symptomatic disease.

[0102] Preferably, post-exposure prophylaxis involves administration of the antibodies disclosed herein after exposure to the coronavirus to prevent severe disease, particularly hospitalization.

[0103] In the treatment methods of the present invention, prophylactic treatment involves administering an antibody to coronavirus at a time when the individual is not infected with coronavirus. Preferably, the antibody binds to a conserved epitope of the spike protein of the coronavirus virion. In some embodiments, the individual in need thereof is not (yet) infected with coronavirus.

[0104] Therapeutic treatment As used herein, the term "therapeutic treatment" includes reference to treating viral infections (including coronavirus diseases) after the viral infection has occurred. Viral infection involves the entry of the virus into the body and / or replication of the virus within the body and / or the spread of the virus to previously uninfected cells, tissues, or locations within the body. Viral infections may cause one or more diseases, but may also remain latent, i.e., remain in the body without causing symptoms or disease.

[0105] coronavirus As used herein, the term "coronavirus" includes reference to positive-sense, single-stranded RNA viruses belonging to the Coronaviridae family.

[0106] Preferably, the antibodies disclosed herein are capable of specifically binding to the stem helix of coronaviruses, particularly SARS-CoV-2.

[0107] Preferably, the antibodies disclosed herein are capable of specifically binding to the stem helix of a SARS-CoV-2 variant of concern.

[0108] Preferably, the antibodies disclosed herein are capable of neutralizing coronaviruses, particularly SARS-CoV-2.

[0109] Preferably, the antibodies disclosed herein are capable of neutralizing at least one or more, preferably two or more, preferably three or more, preferably four or more, and even more preferably five or more coronavirus subtypes.

[0110] As used herein, the phrase "capable of" means that the ability of the antibodies or antigen-binding fragments thereof described herein is sufficient to bind to at least a portion of an antigen, such as the stem helix of a coronavirus, particularly SARS-CoV-2, and / or a fusion protein of a coronavirus, particularly SARS-CoV-2. The binding affinity or binding ability of the antibodies and / or antigen-binding fragments thereof described herein can be measured using any method known to one of skill in the art. As used herein, the phrase "capable of" includes, but is not limited to, the phrases "specifically bind" or "specifically binds to."

[0111] coronavirus infection As used herein, the term "coronavirus infection" includes reference to the pathological or non-pathological, preferably pathological, entry and retention of any type of coronavirus in a human host. The infecting virus may replicate within the host, its cells, or the cells of its microbiota. The infecting virus may or may not cause disease, e.g., COVID-19. The infection may or may not be detectable by methods known in the art for detecting viral infection. An infected individual may or may not be aware of the infection. Typical, but not exclusive, sites in the human body where SARS-CoV-2, for example, may be located in an infected individual are the respiratory system and / or its cells, and the cardiovascular system and / or its cells. As used herein, the term "coronavirus infection" further includes reference to the entry and retention of any type of coronavirus moiety capable of initiating viral replication in a human host. The term "coronavirus infection" encompasses symptoms or disease subsequent to infection, e.g., COVID-19. The term "coronavirus infection" encompasses "SARS-CoV-2 infection."

[0112] epitope As used herein, the term "epitope" includes reference to a moiety capable of binding to an antibody disclosed herein with sufficiently high affinity to form a detectable antigen-antibody complex.

[0113] individual As used herein, the term "individual" includes reference to a mammal or human subject to or at risk of contracting a viral infection. Infection can occur in any system, tissue, or cell belonging to the host, including the host's microbiota. Coronavirus infection and coronavirus disease can occur in individuals of all age groups and genders. Nevertheless, preferably, the individual is a human, particularly an elderly human, e.g., at least 60, 65, 70, 75, 80, or at least 85 years of age, or a human at increased risk of infection due to their occupation or living environment. Preferably, the individual is at risk of contracting a serious disease upon infection, e.g., COVID-19. In preferred embodiments, the individual has an underlying condition, such as (i) a respiratory disease, such as asthma, COPD, chronic bronchitis, and emphysema; (ii) a cardiovascular disease, such as cardiac arrhythmia or an individual who has undergone cardiac surgery; (iii) diabetes; (iv) renal failure; and / or (v) a disease affecting the immune system, e.g., an immunocompromised individual or an increased risk of viral infection due to their occupation.

[0114] As used herein, the term "individual" includes reference to a mammal, such as, but not limited to, a human, that will benefit from a specified therapy; for example, the term "individual" can encompass a non-human primate (NHP).

[0115] Preferably, the individual is a mammal, more preferably a human.

[0116] Administering and Administration As used herein, the terms "administering" and "administration" include reference to providing one or more drugs and optionally one or more adjuvants for the purpose of treating, curing, reducing, or preventing a disease or its symptoms in an individual, or promoting the individual's sense of well-being. Preferred methods of administering the antibodies disclosed herein include transmucosal administration, preferably intranasal administration, and oral inhalation.

[0117] Individuals who need it As used herein, the phrase "individual in need thereof" includes reference to a mammal, such as a human, that will benefit from the identified therapy. The treatment methods of the present invention may be used prophylactically and do not require the presentation of symptoms or signs of coronavirus infection. Individuals particularly in need of the methods or antibodies for use of the present invention are those at high risk of coronavirus infection, at high risk of developing severe symptoms (disease), such as COVID-19, and / or at high risk of dying from COVID-19. Those skilled in the art are familiar with risk factors for high risk of coronavirus infection, high risk of developing severe symptoms of coronavirus infection, and high risk of dying from coronavirus infection.

[0118] antibody The term "antibody," as used herein, includes reference to an intact immunoglobulin, including a monoclonal antibody, e.g., a chimeric, humanized, or human monoclonal antibody, or a binding molecule that comprises an antigen-binding domain (e.g., heavy chain CDRs 1-3 of the variable domain) of an antibody disclosed herein, or an antibody that competes with an antibody disclosed herein for specific binding to an immunoglobulin binding partner.

[0119] In other words, functional fragments of antibodies are also encompassed by the term "antibody." Antibodies are generally Y-shaped proteins. In antibodies, a constant domain and a variable domain are generally present. The variable domain facilitates antigen binding. Antibodies generally comprise two heavy chains and two light chains. Both the heavy and light chains are partly constant and partly variable. Antibodies occur in several classes, namely IgA, IgD, IgE, IgG, and IgM. Preferably, the antibodies of the present invention are of the IgG class, particularly the IgG1 class. Some classes are further subdivided into subclasses or isotypes. For example, the IgG class is subdivided into subclasses IgG1, IgG2, IgG3, and IgG4. Preferably, the antibodies of the present invention are of the IgG class, preferably the IgG1 class. Antigen-binding regions or antigen-binding fragments of antibodies are encompassed by the term "antibody" and are therefore part of the present invention, and may include, for example, Fab, F(ab'), F(ab')2, dAb, Fv, Fd, CDR fragments, diabodies, triabodies, tetrabodies, single-chain antibodies (scFv), scFv-Fc, bivalent single-chain antibodies, single-chain phage antibodies, (poly)peptides comprising at least a fragment of an immunoglobulin sufficient to confer specific antigen-binding properties to the (poly)peptide, and others. Such fragments may be produced synthetically or by enzymatic or chemical cleavage of the original immunoglobulin, or may be genetically engineered by recombinant DNA techniques. Methods for producing antibodies and antigen-binding fragments are well known to those skilled in the art. Antibodies may be conjugated or unconjugated. Antibodies may be conjugated, linked, or otherwise physically or functionally associated with effector moieties or tags, such as, for example, enzymes, liposomes, radioactive substances, fluorescent dyes, toxic substances, among others. Antibodies may be stabilized, multimerized, humanized, or otherwise engineered.

[0120] The antibody may be neutralizing. This includes reference to the inhibition of virus, e.g., as measured by an in vitro neutralization assay of virus entry into host cells and / or virus replication. Neutralization can be achieved, for example, by inhibiting attachment or adhesion of the virus to the cell surface, by inhibiting fusion of the virus with the cell membrane after the virus has attached to the target cell, or by inhibiting virus egress from the cell. Neutralization does not specify the method of neutralization. Preferably, the antibody is cross-neutralizing. This includes reference to the ability of the antibody of the present invention to bind to and neutralize different sets of molecules, preferably different sets of molecules of different subtypes belonging to the Coronaviridae family.

[0121] As used herein, the terms "domain" or "region" in reference to a portion of an antibody, bispecific antibody, or antigen-binding fragment thereof, may be used interchangeably herein.

[0122] As used herein, the phrases "antigen-binding fragment," "antigen-binding fragment," "antigen fragment," "fragment of an antibody," or "antigen-binding molecule" refer to a protein, polypeptide, or molecular complex that comprises or consists of at least one CDR, alone or in combination with one or more additional complementarity-determining regions (CDRs) and / or framework regions (FRs) described herein, that is capable of binding to the stem helix of a coronavirus, particularly SARS-CoV-2, and / or a portion of an antigen comprising the stem helix of a coronavirus, particularly SARS-CoV-2 and its variants of concern. As used herein, the phrases "antigen-binding fragment," "antigen-binding fragment," "antigen fragment," "fragment of an antibody," or "antigen-binding molecule" can be used interchangeably. For example, the phrase "antigen-binding fragment" as used herein is used interchangeably with the phrase "antibody fragment" as used herein.

[0123] Preferably, the antibodies of the invention disclosed herein are capable of cross-neutralizing coronaviruses, particularly SARS-CoV-2.

[0124] Antibodies contain complementarity-determining regions (CDRs) located in the variable domains of the heavy and light chains. CDRs contribute significantly to the antigen-binding site. Three CDRs can be identified: CDR1, CDR2, and CDR3. Because each CDR can be located on either the light or heavy chain, for each antigen receptor, there are generally six CDRs that collectively contact the antigen: light chain CDR1, light chain CDR2, light chain CDR3, heavy chain CDR1, heavy chain CDR2, and heavy chain CDR3. CDR3-type CDRs are the most variable. CDRs can be specific for linear, discontinuous, or conformational epitopes of proteins or protein fragments, whether present on the protein in its native conformation or, in some cases, on denatured or inactivated proteins. Epitopes can also consist of or include post-translational modifications of proteins. Of particular interest are antibodies of the present invention that contain CDRs that recognize coronavirus antigens, such as spike proteins, and in particular the stem helices of spike proteins. The antibodies disclosed herein bind to conserved epitopes in the stem helices at the base of the spike protein of the SARS-CoV-2 virion. The epitopes of the antibodies disclosed herein use the CDR loops of the light and heavy chains.

[0125] The antibodies disclosed herein can be used in isolated or non-isolated form.

[0126] Preferably, the compositions of the present invention comprise a single anti-coronavirus antibody disclosed herein.

[0127] Furthermore, the antibodies disclosed herein can be used alone or in mixtures comprising the antibodies (or variants, fragments, or bispecifics thereof) disclosed herein and / or with other antibodies that bind to coronavirus and have a coronavirus inhibitory effect. In other words, the antibodies disclosed herein can be used in combination, e.g., as pharmaceutical compositions or co-administered compositions comprising two or more antibodies that specifically bind to coronavirus. For example, antibodies with different but complementary activities can be combined in a single therapy to achieve a desired therapeutic or prophylactic effect. Alternatively, antibodies with the same activity can be combined in a single therapy to achieve a desired prophylactic or therapeutic effect. Optionally, the mixture further comprises at least one other therapeutic agent.

[0128] Preferably, the antibodies disclosed herein are bispecific antibodies and / or antigen-binding fragments thereof, and the bispecific antibodies are capable of binding to the stem helix of a coronavirus and / or the fusion peptide of a coronavirus. The phrase "capable of binding" encompasses "specifically binds to," in other words, the antibodies, bispecific antibodies, and / or antigen-binding fragments thereof disclosed herein that are capable of binding to an epitope may also bind to other epitopes.

[0129] Preferably, the coronaviruses described herein include SARS-CoV-2 and variants.

[0130] As used herein, the phrases "replace each other," "interchanged," "swap positions with each other," or "replace one another" include reference to when at least a portion of a heavy chain domain, e.g., CH1 and / or VH, of an antibody or antigen-binding fragment described herein is replaced by at least a portion of the light chain domain with which it is paired, e.g., CL and / or VL, and at the same time, when at least a portion of the light chain domain is replaced by at least a portion of the heavy chain domain. The phrases "replace each other," "interchanged," "swap positions with each other," or "replace one another" are used interchangeably herein.

[0131] Preferably, the antibodies disclosed herein are human antibodies.

[0132] Framework Area Antibodies also contain framework regions, generally four framework regions (FR1, FR2, FR3, and FR4), in each of the variable heavy and light domains. The CDRs are located between the framework regions (preferably in the order FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4), and together the CDRs and framework regions define a heavy chain variable domain (or region) and a light chain variable domain (or region), which are also called antigen-binding regions or domains.

[0133] IgG antibodies As used herein, the term "IgG antibody" includes reference to an antibody that contains two antigen-binding sites. IgG is the most common antibody in human serum. The heavy chain of an IgG antibody is of the γ type, which can be subdivided into γ1, γ2, γ3, and γ4. Preferably, the antibodies described herein are γ1 heavy chains. The light chain of an IgG antibody is of the λ or κ type. Preferably, the antibodies described herein are λ light chains.

[0134] Preferably, the antibody is an IgG antibody, preferably an IgG1 antibody.

[0135] IgA antibody As used herein, the term "IgA antibody" includes reference to antibodies containing two to four antigen-binding sites. IgA is the most abundant antibody in mucosal secretions. IgA antibodies are produced in monomeric, dimeric, or secretory forms, each with two or four antigen-binding sites, and can exist as IgA1 or IgA2 isotypes. The heavy chain of an IgA antibody is of the α type, which can be subdivided into α1 and α2. The light chain of an IgA antibody is of the λ or κ type.

[0136] Preferably, the antibody is an IgA antibody.

[0137] IgM antibody As used herein, the term "IgM antibody" includes reference to antibodies containing 10 to 12 antigen-binding sites. IgM antibodies can be produced in pentameric or hexamer forms. The heavy chains of IgM antibodies are of the μ type. The light chains of IgM antibodies are of the λ or κ type.

[0138] Preferably, the antibody is an IgM antibody.

[0139] Anti-coronavirus antibodies As used herein, the term "anti-coronavirus antibody" includes reference to the antibodies disclosed herein, i.e., antibodies that can be used in the treatment methods of the present invention. Preferably, the anti-coronavirus antibodies of the present invention can treat various subtypes of coronavirus.

[0140] For the avoidance of doubt, the term "anti-coronavirus antibody" can be used interchangeably with "coronavirus antibody."

[0141] composition In preferred embodiments, the antibodies disclosed herein are the only active ingredient in the composition administered for treatment, eg, the antibody is provided in the composition as the only active ingredient.

[0142] In a preferred embodiment, the active ingredient comprises an antibody disclosed herein, e.g., the antibody is provided as a composition as the sole active ingredient in the composition administered for treatment or in combination with another antibody.

[0143] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The CDR sequence includes any one of 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, or SEQ ID NO: 644. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0144] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The first antibody comprises a CDR sequence comprising any one of SEQ ID NOs: 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises a CDR sequence comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, and 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 645, 646, 647, 648, 649, or SEQ ID NO: 650. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0145] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The CDR sequence includes any one of 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 651, 652, 653, 654, 655, or SEQ ID NO: 656. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0146] In a preferred embodiment, a first antibody disclosed herein is provided as a composition in combination with a second antibody, wherein the first antibody is selected from the group consisting of SEQ ID NOs: 001-006, 012-017, 023-028, 033-038, 042-047, 050-055, 077-082, 088-093, 099-104, 110-115, 121-126, 132-137, 197-202, 208-213, 219-224, 230-232, 232-234, 234-236, 236-238, 240-242, 242-246, 244-248, 246-248, 248-249, 250-251, 252-253, 254-255, 256-257, 258-259, 260-261, 262-263, 264-265, 266-267, 268-269, 270-271, 272-273, 274-275, 276-277, 278-279, 280-281, 282-283, 284-285, 286-287, 288-289, 290-291, 300-302, 304-305, 306-307, 308-309, 310-311, 312-313, 314-315, 316 The CDR sequence includes any one of 0 to 235, 241 to 246, 252 to 257, 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, 358 to 363, 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, 475 to 480, 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. More preferably, the first antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second antibody comprises CDR sequences comprising any one or more of SEQ ID NOs: 657, 658, 659, 660, 661, or SEQ ID NO: 662. Preferably, the composition is administered for prophylactic and / or therapeutic treatment.

[0147] Preferably, the compositions of the present invention are water-based compositions, such as aqueous liquids. Preferably, the compositions of the present invention further comprise one or more salts, such as sodium chloride. The compositions of the present invention may further comprise one or more buffering agents, such as sodium acetate. The compositions of the present invention may further comprise one or more carbohydrates, such as sucrose, or other active ingredients, such as other antibodies, neuraminidase inhibitors, endonuclease inhibitors, and adjuvants, such as oils, cytokines, emulsifiers, or combinations thereof. The pH of the compositions of the present invention may be between 4 and 8, and more preferably, the pH is around 5.5. Preferably, the pH is around 7.4.

[0148] Composition for mucous membranes As used herein, the term "mucosal composition" refers to any pharmaceutical preparation that is topically applied to a mucosal surface to retard or resist the effects of mucosal shedding or removal.

[0149] The term "mucosal composition" is used in reference to the route of administration in which the antibodies disclosed herein are provided to the nasal cavity, oropharyngeal region, or upper respiratory tract, preferably through the nasal passages, as part of the prophylactic and / or therapeutic treatments disclosed herein.

[0150] Transmucosal Mucous membranes line the internal cavities of the human body, cover internal organs, and comprise membranes composed of one or more layers of epithelial cells and loose connective tissue, which may produce mucus. As used herein, the term "transmucosal," also referred to as "mucosal administration," refers to a route of administration in which a drug is provided prophylactically and / or therapeutically to a mucous membrane, such as those found in the nose, mouth, lungs, vagina, rectum, or stomach. Mucous membranes lubricate these organs and cavities, protecting them from abrasive particles and fluids, as well as invasive pathogens.

[0151] Preferably, the antibodies of the invention are administered transmucosally.

[0152] Intranasal As used herein, the term "intranasal," also referred to as "nasal administration," includes reference to a route of administration in which a drug is preferably provided to the upper and / or lower respiratory tract through the nostrils as part of the prophylactic and / or therapeutic treatments disclosed herein. Preferably, administration provides the drug to the nasal cavity. The back of the nasal cavity is also referred to as the pharynx. Nasal administration preferably provides delivery of the antibodies disclosed herein to the mucous membranes lining the nasal cavity. Nasal administration can be performed, for example, using a nasal spray or nasal drops. In some embodiments, the drug is delivered to the nasal cavity via the oral route. For example, RetroNose uses a breath-activated pressurized metered-dose inhaler (pMDI) to administer the drug through the oral cavity during the nasal exhalation phase. This method allows drug particles to enter the nasal cavity through the pharynx.

[0153] Preferably, the antibodies of the invention are administered intranasally.

[0154] In nasal administration, a drug is provided to the upper respiratory tract through the nostrils as part of the preventive and / or therapeutic treatments disclosed herein.Preferably, the administration provides the drug to the nasal cavity.Since the drug delivered locally in this way can continue to have a local or systemic effect, nasal administration can be a form of local or systemic administration.In the present case with the antibody disclosed herein, nasal administration is preferably a form of local administration.

[0155] The intranasal administration disclosed herein may be carried out using a medicament in liquid form, preferably in the form of drops or nasal spray. The aqueous liquid may contain an adjuvant. These adjuvants may be, for example, salts, oils, cytokines, emulsifiers, buffers, carbohydrates, and combinations thereof. Nasal administration may also be carried out using a medicament in solid form, such as a powder.

[0156] Oral inhalation As used herein, the term "oral inhalation," also referred to as "mouth inhalation," includes reference to a route of administration in which a drug is provided through the mouth to the upper and / or lower respiratory tract, e.g., the lungs, as part of the prophylactic and / or therapeutic treatment of the invention. Oral inhalation may apply, for example, to drugs in powder form as well as drugs in the form of liquid droplets or aerosols.

[0157] Preferably, the antibodies of the invention are administered by oral inhalation.

[0158] In oral inhalation, a drug is provided through the mouth to the respiratory tract, preferably the lower respiratory tract, e.g., the lungs, as part of the prophylactic and / or therapeutic treatment of the invention. As further described herein, oral inhalation also includes nasal drug delivery (also referred to as nasal drug delivery via the oral route).

[0159] Oral inhalation may be applied to, for example, drugs in powder form and drugs in droplet or aerosol form. Oral inhalation may involve the use of an inhaler. The inhaler may be involved in achieving a determined dose. Drugs administered by oral inhalation may reach the lungs, but may also be partially excreted through exhaled air.

[0160] The oral inhalation administration disclosed herein may be carried out using a medicament containing an aerosol in powder (solid) or liquid form. Powdered aerosols containing particles smaller than 3 μm in diameter will primarily reach the respiratory portion of the lungs and therefore will be better absorbed than larger particles. The medicament may contain adjuvants. These adjuvants may be, for example, salts, oils, cytokines, emulsifiers, buffers, carbohydrates, and combinations thereof.

[0161] Oral pharyngeal administration As used herein, the term "oropharyngeal administration," also referred to as delivery to the portion of the pharynx located between the soft palate and the hyoid bone, includes reference to routes of administration in which a drug is provided via the oral or nasal passages as part of a prophylactic and / or therapeutic treatment. Oropharyngeal administration may be used, for example, for drugs in powder form as well as for drugs in the form of liquid drops or aerosols.

[0162] Preferably, the antibodies of the invention are administered by oral pharyngeal administration.

[0163] in front As used herein, the term "prior" includes reference to administration of the antibody before an individual is exposed to or infected with coronavirus.

[0164] Preferably, the disclosed antibodies are administered to an individual up to 24 hours before exposure to a coronavirus, for example, 0-24 hours before the individual is exposed to said coronavirus.

[0165] Preferably, the disclosed antibodies are administered to an individual up to 48 hours before exposure to a coronavirus, for example, 0-48 hours before the individual is exposed to said coronavirus.

[0166] In a preferred embodiment, the antibody is administered two or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered three or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered four or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered five or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered six or more days prior to exposure to coronavirus. In a preferred embodiment, the antibody is administered seven or more days prior to exposure to coronavirus.

[0167] Dosage As used herein, the term "dosage" refers to an amount of antibody given over a particular period of time (e.g., over the course of 24 hours, 12 hours, 30 minutes, etc.). A dose refers to a single administration episode (e.g., taking two or more tablets, receiving two or more nasal doses), whether the dose is a unit dosage form or a multiple unit dosage form combined together. A dosage includes reference to pharmaceutical dosage forms in which the medication is packaged for administration, e.g., as a single unit dose or multiple unit doses. A dosage may be administered, for example, as one or more drops of an antibody-containing composition (e.g., a nasal spray) or one or more sprays of an antibody-containing composition (e.g., a nasal spray).

[0168] Preferably, suitable dosages of the antibodies disclosed herein, for example, when the dosage is for intranasal administration, include doses of 0.01 mg to 20 mg, preferably 0.1 mg to 15 mg, more preferably about 0.5 mg and 10 mg, or about 1 mg. Such dosages are also referred to as "fixed dosages" or "nominal dosages," as opposed to dosages based on patient weight. Fixed dosages have the advantage that the medicament can be packaged, for example, in a single unit dose. A single dose of the antibodies according to the present invention can provide protection from coronavirus infection for several days and can be provided "on demand" or "as needed." For example, an individual can receive the antibody before leaving home or before coming into contact with other individuals.

[0169] To provide long-lasting protection, the antibody may be administered periodically. For example, the antibody is administered once a month, or at least once a month. In a preferred embodiment, the antibody is administered once a week, or at least once a week, for example, twice a week. In a preferred embodiment, the antibody is administered once a day, or at least once a day. As will be apparent to those skilled in the art, less antibody may be administered if the antibody is administered more frequently (e.g., daily). In a preferred embodiment, 0.01 mg to 20 mg of antibody per week is administered (e.g., once or twice a week or daily). In an exemplary embodiment, 0.1 mg to 25 mg (i.e., 0.7 mg to 175 mg per week) is administered daily. In an exemplary embodiment, 0.5 mg to 3.5 mg (i.e., 3.5 mg to 24.5 mg per week) is administered daily.

[0170] The present invention also provides compositions formulated for intranasal administration and / or oral inhalation comprising an antibody disclosed herein, preferably in a single dosage unit of between 0.1 mg and 20 mg, preferably between 0.5 mg and 15 mg, or preferably between 1 mg and 12.5 mg.

[0171] Use in methods for treatment The present invention particularly relates to the antibodies disclosed herein for use in methods for the treatment of coronavirus infection in an individual. More specifically, the antibodies can be used in methods for the prophylactic and / or therapeutic treatment of coronavirus infection in an individual.

[0172] Complementarity-Determining Regions (CDRs) Preferably, the CDR regions are identified according to Kabat et al. (1991) as described in Sequences of Proteins of Immunological Interest.

[0173] In a preferred embodiment, the binding interaction between the binding molecule, preferably an antibody, and the stem helix is ​​mediated exclusively by the variable sequences of the light and heavy chains.

[0174] The antibodies disclosed herein can specifically bind to coronavirus in attenuated or inactivated form or in live and / or infectious form.

[0175] The antibodies disclosed herein can also specifically bind to one or more fragments of a coronavirus.

[0176] The antibodies disclosed herein preferably comprise heavy chain CDR1 sequences comprising any one of the following SEQ ID NOs: 001 to 006, 012 to 017, 023 to 028, 033 to 038, 042 to 047, and 050 to 055. A CDR comprising the amino acid sequence of SEQ ID NO: 055 is particularly preferred.

[0177] The antibodies disclosed herein preferably comprise a heavy chain CDR2 sequence comprising any one of the following SEQ ID NOs: 077 to 082, 088 to 093, 099 to 104, 110 to 115, 121 to 126, and 132 to 137. A CDR comprising the amino acid sequence of SEQ ID NO: 137 is particularly preferred.

[0178] The antibodies disclosed herein preferably comprise heavy chain CDR3 sequences comprising any one of the following SEQ ID NOs: 197 to 202, 208 to 213, 219 to 224, 230 to 235, 241 to 246, and 252 to 257. A CDR comprising the amino acid sequence of SEQ ID NO: 257 is particularly preferred.

[0179] The antibodies disclosed herein preferably comprise a light chain CDR1 sequence comprising any one of the following SEQ ID NOs: 303 to 308, 314 to 319, 325 to 330, 336 to 341, 347 to 352, and 358 to 363. A CDR comprising the amino acid sequence of SEQ ID NO: 363 is particularly preferred.

[0180] The antibodies disclosed herein preferably comprise a light chain CDR2 sequence comprising any one of the following SEQ ID NOs: 421 to 426, 432 to 437, 443 to 448, 454 to 459, 465 to 470, and 475 to 480. A CDR comprising the amino acid sequence of SEQ ID NO: 480 is particularly preferred.

[0181] The antibodies disclosed herein preferably comprise a light chain CDR3 sequence comprising any one of the following SEQ ID NOs: 514 to 519, 525 to 530, 536 to 541, 547 to 552, 558 to 563, and 569 to 574. A CDR comprising the amino acid sequence of SEQ ID NO: 574 is particularly preferred.

[0182] The antibodies disclosed herein comprise a heavy chain variable domain further comprising at least the heavy chain framework region FR1 of SEQ ID NO:631.

[0183] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the heavy chain framework region FR2 of SEQ ID NO:632.

[0184] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the heavy chain framework region FR3 of SEQ ID NO:633.

[0185] The antibodies disclosed herein comprise a heavy chain variable domain further comprising at least the heavy chain framework region FR4 of SEQ ID NO:634.

[0186] In a preferred embodiment, the heavy chain variable domain of the antibody comprises a heavy chain framework region FR1 of SEQ ID NO: 631, a heavy chain framework region FR2 of SEQ ID NO: 632, a heavy chain framework region FR3 of SEQ ID NO: 633, and / or a heavy chain framework region FR4 of SEQ ID NO: 634, preferably all of the heavy chain framework regions FR1 to FR4.

[0187] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR1 of SEQ ID NO:635.

[0188] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR2 of SEQ ID NO:636.

[0189] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR3 of SEQ ID NO:637.

[0190] The antibodies disclosed herein comprise a heavy chain variable domain that further comprises at least the light chain framework region FR4 of SEQ ID NO:638.

[0191] In a preferred embodiment, the light chain variable domain of the antibody comprises a light chain framework region FR1 of SEQ ID NO: 635, a light chain framework region FR2 of SEQ ID NO: 636, a light chain framework region FR3 of SEQ ID NO: 637, and / or a light chain framework region FR4 of SEQ ID NO: 638, preferably all of the light chain framework regions FR1 to FR4.

[0192] Most preferably, the antibodies disclosed herein comprise a heavy chain variable domain having the sequence of SEQ ID NO:629 and / or a light chain variable domain having the sequence of SEQ ID NO:630.

[0193] In the treatment methods or compositions of the present invention, the heavy chain variable domain (VH) of the antibody is preferably set forth as SEQ ID NO: 629. In the treatment methods or compositions of the present invention, the light chain variable domain (VL) of the antibody is preferably set forth as SEQ ID NO: 630. In the treatment methods or compositions of the present invention, the antibody may be an antibody disclosed herein or a binding molecule comprising an antigen-binding domain thereof, such as a variable domain (V).

[0194] Preferably, the antibodies disclosed herein comprise a heavy chain variable domain having the sequence of SEQ ID NO: 629 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, more preferably 0, 1, 2, 3, 4, or 5 amino acid insertions, deletions, or substitutions.

[0195] Preferably, the antibodies disclosed herein comprise a light chain variable domain having the sequence of SEQ ID NO: 630 with at most 15, preferably 0, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10, more preferably 0, 1, 2, 3, 4, or 5 amino acid insertions, deletions, or substitutions.

[0196] Preferably, said amino acid insertions, deletions or substitutions for the heavy and / or light chain variable domains are not within the CDRs.

[0197] array CDR and framework regions as defined herein are based on the Kabat numbering scheme set forth in Sequences of Proteins of Immunological Interest (Kabat et al., 1991). Heavy chain CDR1 region (SEQ ID NOs: 001 to 076) Sequence number 001 GFTFSYFYLHWVRQA Sequence number 002 FTFSYFYLHWVRQA Sequence number 003 TFSYFYLHWVRQA Sequence number 004 FSYFYLHWVRQA Sequence number 005 SYFYLHWVRQA Sequence number 006 YFYLHWVRQA Sequence number 007 FYLHWVRQA Sequence number 008 YLHWVRQA Sequence number 009 LHWVRQA Sequence number 010 HWVRQA Sequence number 011 WVRQA Sequence number 012 GFTFSYFYLHWVRQ Sequence number 013 FTFSYFYLHWVRQ Sequence number 014 TFSYFYLHWVRQ Sequence number 015 FSYFYLHWVRQ Sequence number 016 SYFYLHWVRQ SEQ ID NO: 017 YFYLHWVRQ Sequence number 018 FYLHWVRQ SEQ ID NO: 019 YLHWVRQ Sequence number 020 LHWVRQ Sequence number 021 HWVRQ SEQ ID NO. 022 WVRQ Sequence number 023 GFTFSYFYLHWVR Sequence number 024 FTFSYFYLHWVR Sequence number 025 TFSYFYLHWVR Sequence number 026 FSYFYLHWVR Sequence number 027 SYFYLHWVR SEQ ID NO: 028 YFYLHWVR Sequence number 029 FYLHWVR Sequence number 030 YLHWVR Sequence number 031 LHWVR Sequence number 032 HWVR SEQ ID NO: 033 GFTFSYFYLHWV SEQ ID NO: 034 FTFSYFYLHWV SEQ ID NO: 035 TFSYFYLHWV SEQ ID NO: 036 FSYFYLHWV SEQ ID NO: 037 SYFYLHWV SEQ ID NO: 038 YFYLHWV Sequence number 039 FYLHWV SEQ ID NO: 040 YLHWV SEQ ID NO. 041 LHWV Sequence number 042 GFTFSYFYLHW Sequence number 043 FTFSYFYLHW Sequence number 044 TFSYFYLHW Sequence number 045 FSYFYLHW Sequence number 046 SYFYLHW SEQ ID NO. 047 YFYLHW Sequence number 048 FYLHW Sequence number 049 YLHW SEQ ID NO: 050 GFTFSYFYLH Sequence number 051 FTFSYFYLH SEQ ID NO: 052 TFSYFYLH SEQ ID NO: 053 FSYFYLH SEQ ID NO: 054 SYFYLH SEQ ID NO: 055 YFYLH Sequence number 056 FYLH SEQ ID NO: 057 GFTFSYFYL SEQ ID NO: 058 FTFSYFYL SEQ ID NO: 059 TFSYFYL Sequence number 060 FSYFYL SEQ ID NO: 061 SYFYL SEQ ID NO: 062 YFYL SEQ ID NO: 063 GFTFSYFY SEQ ID NO: 064 FTFSYFY SEQ ID NO: 065 TFSYFY SEQ ID NO: 066 FSYFY SEQ ID NO: 067 SYFY SEQ ID NO: 068 GFTFSYF Sequence number 069 FTFSYF Sequence number 070 TFSYF SEQ ID NO. 071 FSYF SEQ ID NO: 072 GFTFSY SEQ ID NO: 073 FTFSY SEQ ID NO: 074 TFSY Sequence number 075 GFTFS Sequence number 076 FTFS Heavy chain CDR2 region (SEQ ID NOs: 077 to 196) SEQ ID NO: 077 LEWMGIINPRGDGTRYAQKFQGRVTMT Sequence number 078 EWMGIINPRGDGTRYAQKFQGRVTMT SEQ ID NO: 079 WMGIINPRGDGTRYAQKFQGRVTMT SEQ ID NO: 080 MGIINPRGDGTRYAQKFQGRVTMT SEQ ID NO: 081 GIINPRGDGTRYAQKFQGRVTMT SEQ ID NO: 082 IINPRGDGTRYAQKFQGRVTMT SEQ ID NO: 083 INPRGDGTRYAQKFQGRVTMT SEQ ID NO: 084 NPRGDGTRYAQKFQGRVTMT SEQ ID NO: 085 PRGDGTRYAQKFQGRVTMT SEQ ID NO: 086 RGDGTRYAQKFQGRVTMT SEQ ID NO: 087 GDGTRYAQKFQGRVTMT Sequence number 088 LEWMGIINPRGDGTRYAQKFQGRVTM Sequence number 089 EWMGIINPRGDGTRYAQKFQGRVTM Sequence number 090 WMGIINPRGDGTRYAQKFQGRVTM Sequence number 091 MGIINPRGDGTRYAQKFQGRVTM Sequence number 092 GIINPRGDGTRYAQKFQGRVTM Sequence number 093 IINPRGDGTRYAQKFQGRVTM Sequence number 094 INPRGDGTRYAQKFQGRVTM Sequence number 095 NPRGDGTRYAQKFQGRVTM Sequence number 096 PRGDGTRYAQKFQGRVTM Sequence number 097 RGDGTRYAQKFQGRVTM Sequence number 098 GDGTRYAQKFQGRVTM Sequence number 099 LEWMGIINPRGDGTRYAQKFQGRVT Sequence number 100 EWMGIINPRGDGTRYAQKFQGRVT SEQ ID NO: 101 WMGIINPRGDGTRYAQKFQGRVT SEQ ID NO: 102 MGIINPRGDGTRYAQKFQGRVT SEQ ID NO: 103 GIINPRGDGTRYAQKFQGRVT SEQ ID NO: 104 IINPRGDGTRYAQKFQGRVT Sequence number 105 INPRGDGTRYAQKFQGRVT SEQ ID NO: 106 NPRGDGTRYAQKFQGRVT SEQ ID NO: 107 PRGDGTRYAQKFQGRVT SEQ ID NO: 108 RGDGTRYAQKFQGRVT SEQ ID NO: 109 GDGTRYAQKFQGRVT SEQ ID NO: 110 LEWMGIINPRGDGTRYAQKFQGRV Sequence number 111 EWMGIINPRGDGTRYAQKFQGRV SEQ ID NO: 112 WMGIINPRGDGTRYAQKFQGRV SEQ ID NO: 113 MGIINPRGDGTRYAQKFQGRV SEQ ID NO: 114 GIINPRGDGTRYAQKFQGRV SEQ ID NO: 115 IINPRGDGTRYAQKFQGRV SEQ ID NO: 116 INPRGDGTRYAQKFQGRV SEQ ID NO: 117 NPRGDGTRYAQKFQGRV SEQ ID NO: 118 PRGDGTRYAQKFQGRV SEQ ID NO: 119 RGDGTRYAQKFQGRV SEQ ID NO: 120 GDGTRYAQKFQGRV SEQ ID NO: 121 LEWMGIINPRGDGTRYAQKFQGR Sequence number 122 EWMGIINPRGDGTRYAQKFQGR SEQ ID NO: 123 WMGIINPRGDGTRYAQKFQGR SEQ ID NO: 124 MGIINPRGDGTRYAQKFQGR SEQ ID NO: 125 GIINPRGDGTRYAQKFQGR SEQ ID NO: 126 IINPRGDGTRYAQKFQGR SEQ ID NO: 127 INPRGDGTRYAQKFQGR SEQ ID NO: 128 NPRGDGTRYAQKFQGR SEQ ID NO: 129 PRGDGTRYAQKFQGR SEQ ID NO: 130 RGDGTRYAQKFQGR SEQ ID NO: 131 GDGTRYAQKFQGR SEQ ID NO: 132 LEWMGIINPRGDGTRYAQKFQG Sequence number 133 EWMGIINPRGDGTRYAQKFQG SEQ ID NO: 134 WMGIINPRGDGTRYAQKFQG SEQ ID NO: 135 MGIINPRGDGTRYAQKFQG SEQ ID NO: 136 GIINPRGDGTRYAQKFQG SEQ ID NO: 137 IINPRGDGTRYAQKFQG SEQ ID NO: 138 INPRGDGTRYAQKFQG SEQ ID NO: 139 NPRGDGTRYAQKFQG SEQ ID NO: 140 PRGDGTRYAQKFQG SEQ ID NO: 141 RGDGTRYAQKFQG SEQ ID NO: 142 GDGTRYAQKFQG SEQ ID NO: 143 LEWMGIINPRGDGTRYAQKFQ Sequence number 144 EWMGIINPRGDGTRYAQKFQ SEQ ID NO: 145 WMGIINPRGDGTRYAQKFQ SEQ ID NO: 146 MGIINPRGDGTRYAQKFQ SEQ ID NO: 147 GIINPRGDGTRYAQKFQ SEQ ID NO: 148 IINPRGDGTRYAQKFQ SEQ ID NO: 149 INPRGDGTRYAQKFQ SEQ ID NO: 150 NPRGDGTRYAQKFQ SEQ ID NO: 151 PRGDGTRYAQKFQ SEQ ID NO: 152 RGDGTRYAQKFQ SEQ ID NO: 153 GDGTRYAQKFQ SEQ ID NO: 154 LEWMGIINPRGDGTRYAQKF Sequence number 155 EWMGIINPRGDGTRYAQKF SEQ ID NO: 156 WMGIINPRGDGTRYAQKF SEQ ID NO: 157 MGIINPRGDGTRYAQKF SEQ ID NO: 158 GIINPRGDGTRYAQKF Sequence number 159 IINPRGDGTRYAQKF Sequence number 160 INPRGDGTRYAQKF SEQ ID NO: 161 NPRGDGTRYAQKF SEQ ID NO: 162 PRGDGTRYAQKF SEQ ID NO: 163 RGDGTRYAQKF SEQ ID NO: 164 GDGTRYAQKF Sequence number 165 LEWMGIINPRGDGTRYAQK Sequence number 166 EWMGIINPRGDGTRYAQK SEQ ID NO: 167 WMGIINPRGDGTRYAQK SEQ ID NO: 168 MGIINPRGDGTRYAQK SEQ ID NO: 169 GIINPRGDGTRYAQK SEQ ID NO: 170 IINPRGDGTRYAQK SEQ ID NO: 171 INPRGDGTRYAQK SEQ ID NO: 172 NPRGDGTRYAQK SEQ ID NO: 173 PRGDGTRYAQK SEQ ID NO: 174 RGDGTRYAQK SEQ ID NO: 175 GDGTRYAQK SEQ ID NO: 176 LEWMGIINPRGDGTRYAQ Sequence number 177 EWMGIINPRGDGTRYAQ SEQ ID NO: 178 WMGIINPRGDGTRYAQ SEQ ID NO: 179 MGIINPRGDGTRYAQ SEQ ID NO: 180 GIINPRGDGTRYAQ SEQ ID NO: 181 IINPRGDGTRYAQ SEQ ID NO: 182 INPRGDGTRYAQ SEQ ID NO: 183 NPRGDGTRYAQ SEQ ID NO: 184 PRGDGTRYAQ SEQ ID NO: 185 RGDGTRYAQ SEQ ID NO: 186 GDGTRYAQ SEQ ID NO: 187 LEWMGIINPRGDGTRYA Sequence number 188 EWMGIINPRGDGTRYA SEQ ID NO: 189 WMGIINPRGDGTRYA SEQ ID NO: 190 MGIINPRGDGTRYA SEQ ID NO: 191 GIINPRGDGTRYA SEQ ID NO: 192 IINPRGDGTRYA SEQ ID NO: 193 INPRGDGTRYA SEQ ID NO: 194 NPRGDGTRYA SEQ ID NO: 195 PRGDGTRYA SEQ ID NO: 196 RGDGTRYA Heavy chain CDR3 region (SEQ ID NOs: 197 to 302) SEQ ID NO: 197 YYCARGADHGAFDIWGQGT Sequence number 198 YCARGADHGAFDIWGQGT Sequence number 199 CARGADHGAFDIWGQGT Sequence number 200 ARGADHGAFDIWGQGT Sequence number 201 RGADHGAFDIWGQGT Sequence number 202 GADHGAFDIWGQGT SEQ ID NO: 203 ADHGAFDIWGQGT Sequence number 204 DHGAFDIWGQGT Sequence number 205 HGAFDIWGQGT Sequence number 206 GAFDIWGQGT Sequence number 207 AFDIWGQGT Sequence number 208 YYCARGADHGAFDIWGQG Sequence number 209 YCARGADHGAFDIWGQG SEQ ID NO: 210 CARGADHGAFDIWGQG SEQ ID NO: 211 ARGADHGAFDIWGQG SEQ ID NO: 212 RGADHGAFDIWGQG Sequence number 213 GADHGAFDIWGQG SEQ ID NO: 214 ADHGAFDIWGQG SEQ ID NO: 215 DHGAFDIWGQG SEQ ID NO: 216 HGAFDIWGQG Sequence number 217 GAFDIWGQG SEQ ID NO: 218 AFDIWGQG SEQ ID NO: 219 YYCARGADHGAFDIWGQ SEQ ID NO: 220 YCARGADHGAFDIWGQ SEQ ID NO: 221 CARGADHGAFDIWGQ SEQ ID NO: 222 ARGADHGAFDIWGQ SEQ ID NO: 223 RGADHGAFDIWGQ Sequence number 224 GADHGAFDIWGQ SEQ ID NO: 225 ADHGAFDIWGQ SEQ ID NO: 226 DHGAFDIWGQ SEQ ID NO: 227 HGAFDIWGQ SEQ ID NO: 228 GAFDIWGQ SEQ ID NO: 229 AFDIWGQ Sequence number 230 YYCARGADHGAFDIWG SEQ ID NO: 231 YCARGADHGAFDIWG SEQ ID NO: 232 CARGADHGAFDIWG SEQ ID NO: 233 ARGADHGAFDIWG SEQ ID NO: 234 RGADHGAFDIWG Sequence number 235 GADHGAFDIWG SEQ ID NO: 236 ADHGAFDIWG Sequence number 237 DHGAFDIWG SEQ ID NO: 238 HGAFDIWG Sequence number 239 GAFDIWG SEQ ID NO: 240 AFDIWG SEQ ID NO: 241 YYCARGADHGAFDIW SEQ ID NO: 242 YCARGADHGAFDIW SEQ ID NO: 243 CARGADHGAFDIW SEQ ID NO: 244 ARGADHGAFDIW SEQ ID NO: 245 RGADHGAFDIW SEQ ID NO: 246 GADHGAFDIW SEQ ID NO: 247 ADHGAFDIW SEQ ID NO: 248 DHGAFDIW SEQ ID NO: 249 HGAFDIW Sequence number 250 GAFDIW Sequence number 251 AFDIW SEQ ID NO: 252 YYCARGADHGAFDI SEQ ID NO: 253 YCARGADHGAFDI SEQ ID NO: 254 CARGADHGAFDI SEQ ID NO: 255 ARGADHGAFDI SEQ ID NO: 256 RGADHGAFDI SEQ ID NO: 257 GADHGAFDI SEQ ID NO: 258 ADHGAFDI SEQ ID NO: 259 DHGAFDI SEQ ID NO: 260 HGAFDI SEQ ID NO. 261 GAFDI SEQ ID NO. 262 AFDI SEQ ID NO: 263 YYCARGADHGAFD SEQ ID NO: 264 YCARGADHGAFD SEQ ID NO: 265 CARGADHGAFD SEQ ID NO: 266 ARGADHGAFD SEQ ID NO: 267 RGADHGAFD SEQ ID NO: 268 GADHGAFD SEQ ID NO: 269 ADHGAFD SEQ ID NO: 270 DHGAFD SEQ ID NO: 271 HGAFD SEQ ID NO: 272 GAFD SEQ ID NO: 273 YYCARGADHGAF SEQ ID NO: 274 YCARGADHGAF SEQ ID NO: 275 CARGADHGAF SEQ ID NO: 276 ARGADHGAF SEQ ID NO: 277 RGADHGAF SEQ ID NO: 278 GADHGAF SEQ ID NO: 279 ADHGAF Sequence number 280 DHGAF SEQ ID NO: 281 HGAF SEQ ID NO: 282 YYCARGADHGA SEQ ID NO: 283 YCARGADHGA SEQ ID NO: 284 CARGADHGA SEQ ID NO: 285 ARGADHGA SEQ ID NO: 286 RGADHGA Sequence number 287 GADHGA SEQ ID NO: 288 ADHGA SEQ ID NO: 289 DHGA SEQ ID NO: 290 YYCARGADHG SEQ ID NO: 291 YCARGADHG SEQ ID NO: 292 CARGADHG SEQ ID NO: 293 ARGADHG SEQ ID NO: 294 RGADHG Sequence number 295 GADHG SEQ ID NO: 296 ADHG SEQ ID NO: 297 YYCARGADH SEQ ID NO: 298 YCARGADH SEQ ID NO: 299 CARGADH Sequence number 300 ARGADH SEQ ID NO: 301 RGADH SEQ ID NO: 302 GADH Light chain CDR1 region (SEQ ID NOs: 303 to 420) SEQ ID NO: 303 ATLSCRASQSVRRNYFAWYQQK SEQ ID NO: 304 TLSCRASQSVRRNYFAWYQQK Sequence number 305 LSCRASQSVRRNYFAWYQQK SEQ ID NO: 306 SCRASQSVRRNYFAWYQQK SEQ ID NO: 307 CRASQSVRRNYFAWYQQK Sequence number 308 RASQSVRRNYFAWYQQK Sequence number 309 ASQSVRRNYFAWYQQK SEQ ID NO: 310 SQSVRRNYFAWYQQK SEQ ID NO: 311 QSVRRNYFAWYQQK SEQ ID NO: 312 SVRRNYFAWYQQK Sequence number 313 VRRNYFAWYQQK SEQ ID NO: 314 ATLSCRASQSVRRNYFAWYQQ SEQ ID NO: 315 TLSCRASQSVRRNYFAWYQQ SEQ ID NO: 316 LSCRASQSVRRNYFAWYQQ SEQ ID NO: 317 SCRASQSVRRNYFAWYQQ SEQ ID NO: 318 CRASQSVRRNYFAWYQQ SEQ ID NO: 319 RASQSVRRNYFAWYQQ SEQ ID NO: 320 ASQSVRRNYFAWYQQ SEQ ID NO: 321 SQSVRRNYFAWYQQ SEQ ID NO: 322 QSVRRNYFAWYQQ SEQ ID NO: 323 SVRRNYFAWYQQ SEQ ID NO: 324 VRRNYFAWYQQ SEQ ID NO: 325 ATLSCRASQSVRRNYFAWYQ SEQ ID NO: 326 TLSCRASQSVRRNYFAWYQ SEQ ID NO: 327 LSCRASQSVRRNYFAWYQ SEQ ID NO: 328 SCRASQSVRRNYFAWYQ SEQ ID NO: 329 CRASQSVRRNYFAWYQ SEQ ID NO: 330 RASQSVRRNYFAWYQ SEQ ID NO: 331 ASQSVRRNYFAWYQ SEQ ID NO:332 SQSVRRNYFAWYQ SEQ ID NO: 333 QSVRRNYFAWYQ SEQ ID NO: 334 SVRRNYFAWYQ Sequence number 335 VRRNYFAWYQ SEQ ID NO: 336 ATLSCRASQSVRRNYFAWY SEQ ID NO: 337 TLSCRASQSVRRNYFAWY SEQ ID NO: 338 LSCRASQSVRRNYFAWY SEQ ID NO: 339 SCRASQSVRRNYFAWY SEQ ID NO: 340 CRASQSVRRNYFAWY SEQ ID NO:341 RASQSVRRNYFAWY SEQ ID NO:342 ASQSVRRNYFAWY SEQ ID NO:343 SQSVRRNYFAWY SEQ ID NO:344 QSVRRNYFAWY SEQ ID NO: 345 SVRRNYFAWY SEQ ID NO:346 VRRNYFAWY SEQ ID NO: 347 ATLSCRASQSVRRNYFAW SEQ ID NO: 348 TLSCRASQSVRRNYFAW SEQ ID NO: 349 LSCRASQSVRRNYFAW SEQ ID NO: 350 SCRASQSVRRNYFAW SEQ ID NO: 351 CRASQSVRRNYFAW SEQ ID NO: 352 RASQSVRRNYFAW SEQ ID NO: 353 ASQSVRRNYFAW SEQ ID NO:354 SQSVRRNYFAW Sequence number 355 QSVRRNYFAW SEQ ID NO: 356 SVRRNYFAW Sequence number 357 VRRNYFAW SEQ ID NO: 358 ATLSCRASQSVRRNYFA SEQ ID NO: 359 TLSCRASQSVRRNYFA SEQ ID NO: 360 LSCRASQSVRRNYFA SEQ ID NO: 361 SCRASQSVRRNYFA SEQ ID NO: 362 CRASQSVRRNYFA SEQ ID NO: 363 RASQSVRRNYFA SEQ ID NO: 364 ASQSVRRNYFA SEQ ID NO: 365 SQSVRRNYFA SEQ ID NO:366 QSVRRNYFA SEQ ID NO: 367 SVRRNYFA SEQ ID NO: 368 VRRNYFA SEQ ID NO: 369 ATLSCRASQSVRRNYF SEQ ID NO: 370 TLSCRASQSVRRNYF SEQ ID NO: 371 LSCRASQSVRRNYF SEQ ID NO: 372 SCRASQSVRRNYF SEQ ID NO: 373 CRASQSVRRNYF SEQ ID NO: 374 RASQSVRRNYF SEQ ID NO: 375 ASQSVRRNYF SEQ ID NO:376 SQSVRRNYF SEQ ID NO:377 QSVRRNYF SEQ ID NO: 378 SVRRNYF Sequence number 379 VRRNYF SEQ ID NO: 380 ATLSCRASQSVRRNY SEQ ID NO:381 TLSCRASQSVRRNY SEQ ID NO: 382 LSCRASQSVRRNY SEQ ID NO: 383 SCRASQSVRRNY SEQ ID NO: 384 CRASQSVRRNY SEQ ID NO: 385 RASQSVRRNY SEQ ID NO: 386 ASQSVRRNY SEQ ID NO:387 SQSVRRNY SEQ ID NO: 388 QSVRRNY SEQ ID NO: 389 SVRRNY SEQ ID NO: 390 VRRNY SEQ ID NO: 391 ATLSCRASQSVRRN SEQ ID NO: 392 TLSCRASQSVRRN SEQ ID NO: 393 LSCRASQSVRRN SEQ ID NO: 394 SCRASQSVRRN SEQ ID NO: 395 CRASQSVRRN SEQ ID NO: 396 RASQSVRRN SEQ ID NO: 397 ASQSVRRN Sequence number 398 SQSVRRN Sequence number 399 QSVRRN Sequence number 400 SVRRN Sequence number 401 VRRN SEQ ID NO: 402 ATLSCRASQSVRR SEQ ID NO: 403 TLSCRASQSVRR SEQ ID NO: 404 LSCRASQSVRR SEQ ID NO: 405 SCRASQSVRR SEQ ID NO: 406 CRASQSVRR SEQ ID NO: 407 RASQSVRR SEQ ID NO: 408 ASQSVRR SEQ ID NO: 409 SQSVRR SEQ ID NO: 410 QSVRR SEQ ID NO: 411 SVRR SEQ ID NO: 412 ATLSCRASQSVR SEQ ID NO: 413 TLSCRASQSVR SEQ ID NO: 414 LSCRASQSVR SEQ ID NO: 415 SCRASQSVR SEQ ID NO: 416 CRASQSVR SEQ ID NO: 417 RASQSVR SEQ ID NO: 418 ASQSVR Sequence number 419 SQSVR SEQ ID NO: 420 QSVR Light chain CDR2 region (SEQ ID NOs: 421 to 513) SEQ ID NO: 421 RLLIYDASTRATGIPDR SEQ ID NO: 422 LLIYDASTRATGIPDR SEQ ID NO: 423 LIYDASTRATGIPDR SEQ ID NO: 424 IYDASTRATGIPDR SEQ ID NO: 425 YDASTRATGIPDR SEQ ID NO: 426 DASTRATGIPDR SEQ ID NO: 427 ASTRATGIPDR SEQ ID NO: 428 STRATGIPDR SEQ ID NO: 429 TRATGIPDR SEQ ID NO: 430 RATGIPDR SEQ ID NO: 431 ATGIPDR SEQ ID NO: 432 RLLIYDASTRATGIPD SEQ ID NO: 433 LLIYDASTRATGIPD SEQ ID NO: 434 LIYDASTRATGIPD SEQ ID NO: 435 IYDASTRATGIPD SEQ ID NO: 436 YDASTRATGIPD SEQ ID NO: 437 DASTRATGIPD SEQ ID NO: 438 ASTRATGIPD SEQ ID NO: 439 STRATGIPD SEQ ID NO: 440 TRATGIPD SEQ ID NO: 441 RATGIPD SEQ ID NO: 442 ATGIPD SEQ ID NO: 443 RLLIYDASTRATGIP SEQ ID NO: 444 LLIYDASTRATGIP SEQ ID NO: 445 LIYDASTRATGIP SEQ ID NO: 446 IYDASTRATGIP SEQ ID NO: 447 YDASTRATGIP SEQ ID NO: 448 DASTRATGIP SEQ ID NO: 449 ASTRATGIP SEQ ID NO: 450 STRATGIP SEQ ID NO: 451 TRATGIP SEQ ID NO: 452 RATGIP SEQ ID NO: 453 ATGIP SEQ ID NO: 454 RLLIYDASTRATGI SEQ ID NO: 455 LLIYDASTRATGI SEQ ID NO: 456 LIYDASTRATGI SEQ ID NO: 457 IYDASTRATGI SEQ ID NO: 458 YDASTRATGI SEQ ID NO: 459 DASTRATGI SEQ ID NO: 460 ASTRATGI SEQ ID NO: 461 STRATGI SEQ ID NO: 462 TRATGI SEQ ID NO: 463 RATGI SEQ ID NO: 464 ATGI SEQ ID NO: 465 RLLIYDASTRATG SEQ ID NO: 466 LLIYDASTRATG SEQ ID NO: 467 LIYDASTRATG SEQ ID NO: 468 IYDASTRATG SEQ ID NO: 469 YDASTRATG SEQ ID NO: 470 DASTRATG SEQ ID NO: 471 ASTRATG SEQ ID NO: 472 STRATG SEQ ID NO: 473 TRATG SEQ ID NO: 474 RATG SEQ ID NO: 475 RLLIYDASTRAT SEQ ID NO: 476 LLIYDASTRAT SEQ ID NO: 477 LIYDASTRAT SEQ ID NO: 478 IYDASTRAT SEQ ID NO: 479 YDASTRAT SEQ ID NO: 480 DASTRAT Sequence number 481 ASTRAT SEQ ID NO: 482 STRAT SEQ ID NO: 483 TRAT SEQ ID NO: 484 RLLIYDASTRA Sequence number 485 LLIYDASTRA Sequence number 486 LIYDASTRA Sequence number 487 IYDASTRA Sequence number 488 YDASTRA Sequence number 489 DASTRA Sequence number 490 ASTRA Sequence number 491 STRA SEQ ID NO: 492 RLLIYDASTR SEQ ID NO. 493 LLIYDASTR SEQ ID NO: 494 LIYDASTR SEQ ID NO. 495 IYDASTR SEQ ID NO: 496 YDASTR SEQ ID NO: 497 DASTR Sequence number 498 ASTR Sequence number 499 RLLIYDAST Sequence number 500 LLIYDAST Sequence number 501 LIYDAST Sequence number 502 IYDAST SEQ ID NO: 503 YDAST SEQ ID NO:504 DAST Sequence number 505 RLLIYDAS Sequence number 506 LLIYDAS Sequence number 507 LIYDAS Sequence number 508 IYDAS Sequence number 509 YDAS SEQ ID NO: 510 RLLIYDA SEQ ID NO:511 LLIYDA SEQ ID NO:512 LIYDA SEQ ID NO: 513 IYDA Light chain CDR3 region (SEQ ID NOs: 514 to 628) SEQ ID NO: 514 AVYYCQQYDSSPPMYIFGQGT SEQ ID NO: 515 VYYCQQYDSSPPMYIFGQGT SEQ ID NO: 516 YYCQQYDSSPPMYIFGQGT SEQ ID NO: 517 YCQQYDSSPPMYIFGQGT SEQ ID NO: 518 CQQYDSSPPMYIFGQGT Sequence number 519 QQYDSSPPMYIFGQGT SEQ ID NO: 520 QYDSSPPMYIFGQGT SEQ ID NO: 521 YDSSPPMYIFGQGT SEQ ID NO: 522 DSSPPMYIFGQGT SEQ ID NO: 523 SSPPMYIFGQGT SEQ ID NO: 524 SPPMYIFGQGT SEQ ID NO: 525 AVYYCQQYDSSPPMYIFGQG SEQ ID NO: 526 VYYCQQYDSSPPMYIFGQG SEQ ID NO: 527 YYCQQYDSSPPMYIFGQG SEQ ID NO: 528 YCQQYDSSPPMYIFGQG SEQ ID NO:529 CQQYDSSPPMYIFGQG SEQ ID NO: 530 QQYDSSPPMYIFGQG SEQ ID NO:531 QYDSSPPMYIFGQG SEQ ID NO: 532 YDSSPPMYIFGQG SEQ ID NO: 533 DSSPPMYIFGQG SEQ ID NO:534 SSPPMYIFGQG SEQ ID NO:535 SPPMYIFGQG SEQ ID NO:536 AVYYCQQYDSSPPMYIFGQ SEQ ID NO:537 VYYCQQYDSSPPMYIFGQ SEQ ID NO: 538 YYCQQYDSSPPMYIFGQ SEQ ID NO: 539 YCQQYDSSPPMYIFGQ SEQ ID NO: 540 CQQYDSSPPMYIFGQ SEQ ID NO:541 QQYDSSPPMYIFGQ SEQ ID NO:542 QYDSSPPMYIFGQ SEQ ID NO: 543 YDSSPPMYIFGQ SEQ ID NO:544 DSSPPMYIFGQ SEQ ID NO:545 SSPPMYIFGQ SEQ ID NO:546 SPPMYIFGQ SEQ ID NO:547 AVYYCQQYDSSPPMYIFG SEQ ID NO:548 VYYCQQYDSSPPMYIFG SEQ ID NO:549 YYCQQYDSSPPMYIFG SEQ ID NO: 550 YCQQYDSSPPMYIFG SEQ ID NO:551 CQQYDSSPPMYIFG SEQ ID NO:552 QQYDSSPPMYIFG SEQ ID NO:553 QYDSSPPMYIFG SEQ ID NO:554 YDSSPPMYIFG SEQ ID NO:555 DSSPPMYIFG SEQ ID NO:556 SSPPMYIFG SEQ ID NO:557 SPPMYIFG SEQ ID NO:558 AVYYCQQYDSSPPMYIF Sequence number 559 VYYCQQYDSSPPMYIF Sequence number 560 YYCQQYDSSPPMYIF SEQ ID NO: 561 YCQQYDSSPPMYIF SEQ ID NO:562 CQQYDSSPPMYIF Sequence number 563 QQYDSSPPMYIF SEQ ID NO:564 QYDSSPPMYIF SEQ ID NO: 565 YDSSPPMYIF SEQ ID NO:566 DSSPPMYIF SEQ ID NO:567 SSPPMYIF SEQ ID NO:568 SPPMYIF SEQ ID NO:569 AVYYCQQYDSSPPMYI SEQ ID NO:570 VYYCQQYDSSPPMYI SEQ ID NO:571 YYCQQYDSSPPMYI SEQ ID NO:572 YCQQYDSSPPMYI SEQ ID NO:573 CQQYDSSPPMYI SEQ ID NO:574 QQYDSSPPMYI SEQ ID NO:575 QYDSSPPMYI SEQ ID NO:576 YDSSPPMYI SEQ ID NO:577 DSSPPMYI SEQ ID NO:578 SSPPMYI SEQ ID NO:579 SPPMYI SEQ ID NO:580 AVYYCQQYDSSPPMY SEQ ID NO:581 VYYCQQYDSSPPMY SEQ ID NO:582 YYCQQYDSSPPMY SEQ ID NO:583 YCQQYDSSPPMY SEQ ID NO:584 CQQYDSSPPMY Sequence number 585 QQYDSSPPMY SEQ ID NO:586 QYDSSPPMY SEQ ID NO:587 YDSSPPMY SEQ ID NO:588 DSSPPMY SEQ ID NO:589 SSPPMY SEQ ID NO:590 SPPMY SEQ ID NO:591 AVYYCQQYDSSPPM SEQ ID NO:592 VYYCQQYDSSPPM SEQ ID NO: 593 YYCQQYDSSPPM SEQ ID NO: 594 YCQQYDSSPPM SEQ ID NO:595 CQQYDSSPPM SEQ ID NO:596 QQYDSSPPM SEQ ID NO:597 QYDSSPPM SEQ ID NO: 598 YDSSPPM SEQ ID NO: 599 DSSPPM Sequence number 600 SSPPM SEQ ID NO: 601 SPPM SEQ ID NO: 602 AVYYCQQYDSSPP Sequence number 603 VYYCQQYDSSPP SEQ ID NO: 604 YYCQQYDSSPP SEQ ID NO: 605 YCQQYDSSPP SEQ ID NO: 606 CQQYDSSPP Sequence number 607 QQYDSSPP SEQ ID NO: 608 QYDSSPP SEQ ID NO: 609 YDSSPP SEQ ID NO: 610 DSSPP SEQ ID NO: 611 SSPP SEQ ID NO: 612 AVYYCQQYDSSP SEQ ID NO: 613 VYYCQQYDSSP SEQ ID NO: 614 YYCQQYDSSP SEQ ID NO: 615 YCQQYDSSP SEQ ID NO: 616 CQQYDSSP SEQ ID NO: 617 QQYDSSP SEQ ID NO: 618 QYDSSP SEQ ID NO: 619 YDSSP SEQ ID NO: 620 DSSP SEQ ID NO: 621 AVYYCQQYDSS Sequence number 622 VYYCQQYDSS SEQ ID NO: 623 YYCQQYDSS SEQ ID NO: 624 YCQQYDSS Sequence number 625 CQQYDSS SEQ ID NO: 626 QQYDSS SEQ ID NO: 627 QYDSS SEQ ID NO: 628 YDSS Heavy Chain Variable Domain (SEQ ID NO: 629) QEQLVQSGAEVKKPGASVKVSCKSSGFTFSYFYLHWVRQAPGQGLEWMGIINPRGDGTRYAQKFQGRVTMTRDASTGTLYMELRSLRSEDTAVYYCARGADHGAFDIWGQGTMVTVSS Light Chain Variable Domain (SEQ ID NO: 630) EIVLTQSPGTLSLSPGERATLSCRASQSVRRNYFAWYQQKRGQAPRLLIYDASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYDSSPPMYIFGQGTKLEIK Heavy chain FR1 region (SEQ ID NO: 631) SEQ ID NO: 631 QEQLVQSGAEVKKPGASVKVSCKSSGFTFS Heavy chain FR2 region (SEQ ID NO: 632) Sequence number 632 WVRQAPGQGLEWMG Heavy chain FR3 region (SEQ ID NO: 633) SEQ ID NO: 633 RVTMTRDASTGTLYMELRSLRSEDTAVYYCAR Heavy chain FR4 region (SEQ ID NO: 634) Sequence number 634 WGQGTMVTVSS Light chain FR1 region (SEQ ID NO: 635) SEQ ID NO: 635 EIVLTQSPGTLSLSPGERATLSC Light chain FR2 region (SEQ ID NO: 636) SEQ ID NO: 636 WYQQKRGQAPRLLIY Light chain FR3 region (SEQ ID NO: 637) SEQ ID NO: 637 GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC Light chain FR4 region (SEQ ID NO: 638) Sequence number 638 FGQGTKLEIK Heavy chain CDR1 region (SEQ ID NO: 639) SEQ ID NO: 639 DYRIH Heavy chain CDR2 region (SEQ ID NO: 640) Sequence number 640 RMNPKSGDTNFAQKFQG Heavy chain CDR3 region (SEQ ID NO: 641) SEQ ID NO: 641 LLIVGGFDPLDDFEV Light chain CDR1 region (SEQ ID NO: 642) SEQ ID NO: 642 SGTSSDVGGYNFVS Light chain CDR2 region (SEQ ID NO: 643) SEQ ID NO: 643 EVTKRPS Light chain CDR3 region (SEQ ID NO: 644) SEQ ID NO: 644 SSYGGTNNLL Heavy chain CDR1 region (SEQ ID NO: 645) SEQ ID NO: 645 GYAMH Heavy chain CDR2 region (SEQ ID NO: 646) SEQ ID NO: 646 VISRDARNKYYADSVKG Heavy chain CDR3 region (SEQ ID NO: 647) SEQ ID NO: 647 LIIPGITEPGSPDALDI Light chain CDR1 region (SEQ ID NO: 648) Sequence number 648 RASQDISKWLA Light chain CDR2 region (SEQ ID NO: 649) SEQ ID NO: 649 AASSLQS Light chain CDR3 region (SEQ ID NO: 650) Sequence number 650 QQASSFPWSIT Heavy chain CDR1 region (SEQ ID NO: 651) SEQ ID NO: 651 SHYMH Heavy chain CDR2 region (SEQ ID NO: 652) Sequence number 652 IINPSGSGTAYGQKFQG Heavy chain CDR3 region (SEQ ID NO: 653) SEQ ID NO: 653 GSGGLFAY Light chain CDR1 region (SEQ ID NO: 654) SEQ ID NO: 654 RASQIVRSNYLA Light chain CDR2 region (SEQ ID NO: 655) Sequence number 655 GASSRAT Light chain CDR3 region (SEQ ID NO: 656) Sequence number 656 LQYDSSPPTYI Heavy chain CDR1 region (SEQ ID NO: 657) SEQ ID NO: 657 SYYMH Heavy chain CDR2 region (SEQ ID NO: 658) Sequence number 658 LITPSGDDTYYAQRFQG Heavy chain CDR3 region (SEQ ID NO: 659) SEQ ID NO: 659 MSRAGGFDV Light chain CDR1 region (SEQ ID NO: 660) SEQ ID NO: 660 RASQSITGRYLA Light chain CDR2 region (SEQ ID NO: 661) SEQ ID NO: 661 GESSRVT Light chain CDR3 region (SEQ ID NO: 662) Sequence number 662 QHFASSPPTYT Heavy chain arm (SEQ ID NO: 663) SEQ ID NO: 663 QEQLVQSGAEVKKPGASVKVSCKSSGFTFSYFYLHWVRQAPGQGLEWMGIINPRGDGTRYAQKFQGRVTMTRDASTGTLYMELRSLRSEDTAVYYCARGADHGAFDIWGQGTM VTVSSASVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECD KTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain arm (SEQ ID NO: 664) SEQ ID NO: 664 EIVLTQSPGTLSLSPGERATLSCRASQSVRRNYFAWYQQKRGQAPRLLIYDASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYDSSPPMYIFGQGTKL EIKSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC Heavy chain arm (SEQ ID NO: 665) SEQ ID NO: 665 QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGFDPLDDFEV WGQGTMVTISSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC DKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIE KTISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain arm (SEQ ID NO: 666) SEQ ID NO: 666 QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKLT VLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS Heavy chain arm (SEQ ID NO: 667) SEQ ID NO: 667 QEQLVQSGAEVKKPGASVKVSCKSSGFTFSYFYLHWVRQAPGQGLEWMGIINPRGDGTRYAQKFQGRVTMTRDASTGTLYMELRSLRSEDTAVYYCARGADHGAFDIWGQGT MVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKT HTCPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKT ISKAKGQPREPQVYTLPPCRDELTKNQVSLWCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain arm (SEQ ID NO: 668) SEQ ID NO: 668 EIVLTQSPGTLSLSPGERATLSCRASQSVRRNYFAWYQQKRGQAPRLLIYDASTRATGIPDRFSGSGSGTDFTLTISRLEPEDFAVYYCQQYDSSPPMYIFGQGTKLE IKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC Heavy chain arm (SEQ ID NO: 669) SEQ ID NO: 669 QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGFDPLDDFEVW GQGTMVTISGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTEC SDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALAPIE KTISKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Light chain arm (SEQ ID NO: 670) SEQ ID NO: 670 QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKL TVLSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC Heavy chain arm (SEQ ID NO: 671) SEQ ID NO: 671 QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKLTVL GQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECSDKTHT CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTI SKAKGQPREPQVCTLPPSRDELTKNQVSLSCAVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLVSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK Arm light chain (SEQ ID NO: 672) SEQ ID NO: 672 QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGFDPLDDFEV WGQGTMVTISSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSC Heavy Chain Variable Domain (SEQ ID NO: 673) QMQLMQSGAEVKKPGASVTVSCKASGDTFSDYRIHWVRQAPGQGLEWMGRMNPKSGDTNFAQKFQGRVTMTRDMSINTAYMTLSGLTFDDTALYYCASLLIVGGDPLDDFEVWGQGTMVTISS Light Chain Variable Domain (SEQ ID NO: 674) QSALTQPPSASGSPGQSVTISCSGTSSDVGGYNFVSWYQHHPGKAPKILIYEVTKRPSGVPDRFSGSKSGNTASLTVSGLQAEDEADYYCSSYGGTNNLLFGGGTKLTVL Heavy Chain Variable Domain (SEQ ID NO: 675) QVQLVESGGGVVQPGRSLRLSCAASGLTFSGYAMHWVRQAPGKGLEWVAVISRDARNKYYADSVKGRFTISRDNSKKTVYLEMNSLRVEDTAVYYCAILIIPGITEPGSPDALDIWGQGTMVSVSS Light Chain Variable Domain (SEQ ID NO: 676) DIQMTQSPSSMSASVGDRVTITCRASQDISKWLAWYQQRPGKAPKLLIYAASSLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQASSFPWSITFGQGTRLEIR [Example]

[0198] The monoclonal antibody CV3-25 specifically binds to the stem helix of SARS-CoV-2, a binding region shared with the antibodies of the present invention, and has demonstrated in vitro protection against the original Wuhan strain of SARS-CoV-2. When administered mucosally at low dosages, CV3-25 provides a lower level of protection compared to its systemic administration. In contrast, when administered mucosally, the antibodies of the present invention provide a greater level of protection, as demonstrated by the in vivo studies presented below.

[0199] Example 1 CV3-25 The objective of this study was to evaluate the pre-exposure efficacy of monoclonal antibody CV3-25 after systemic intraperitoneal administration in a SARS-CoV-2 delta K18 hACE2 Tg mouse model.

[0200] On day -1 of the study, animals (n=10 per group) were treated intraperitoneally with a range of doses of test CV3-25 (15-0.2 mg / kg) or PBS control as vehicle. On day 0 of the study, animals were administered 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0201] Materials and Methods The monoclonal antibody CV3-25 was dissolved and diluted in phosphate buffered saline (PBS) to prepare a CV3-25 dilution.

[0202] Dilutions of CV3-25 were made at various concentrations so that administration of 200 μL resulted in doses of CV3-25 ranging from 15 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg relative to the mean body weight of the dosing group.

[0203] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0204] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0205] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ Stock Number: 034860) mouse model, weighing approximately 17-26 g on day -9. Sixty mice, 11-13 weeks old on the day of test material administration, were used, with 83.3% female. Ten animals were randomly assigned to six treatment groups based on their body weight on day -9 to create groups with similar average body weights. Males comprised 20% of the animals per group, except for the 15 mg / kg group, which was all female. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets and acidified water for animal nutrition, and red plastic tunnels. Each cage system housed 3-5 mice per cage. All mice were moved to new cages every 14 days. Food and water were checked daily and replenished as needed.

[0206] Study design The dose levels of CV3-25 administered in this example were based on the dose range proven to have prophylactic activity. A total of 60 mice, 11-13 weeks old, were transported to the animal facility and assigned to six experimental groups according to Table 1 (see below). Mice were allowed more than 3 days for acclimatization.

[0207] [Table 1]

[0208] SARS-CoV-2 Delta K18 hACE2 mice (20% males per group, except for the 15 mg / kg group, which were all female) were treated via the intraperitoneal route with antibody at doses ranging from 15 mg / kg to 0.2 mg / kg, based on the mean group weight on Day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on Day -1). On Day 0, all mice received a lethal dose (10 3.5 TCID 50) and monitored for weight loss and mortality until the study was terminated on day 11.

[0209] Antibody administration Upon arrival, the test antibodies were stored at -80°C ± 10°C. The appropriate dose according to the treatment schedule (Table 1) was prescribed according to the mean group weight on day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on day -1).

[0210] Prior to dosing, the material was drawn up into a 1 mL syringe with a 26 G needle, briefly warmed to room temperature, and then administered to each mouse. Mice received the indicated dose by intraperitoneal administration of 200 μL of antibody solution into the peritoneal cavity. For administration, mice were grasped by the neck and held upside down at a -45° angle (200 μL per mouse).

[0211] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in sterile PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0212] Laboratory analysis The administered virus dose was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0213] Clinical monitoring General health observations were performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal was weighed daily, beginning on the day of infection (day 0).

[0214] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 11. No complete necropsy was performed.

[0215] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 11 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0216] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0217] survival rate Prophylactic treatment with test antibody at 0.5 mg / kg or greater provided a statistically significant increase in survival compared to the control group (see Figure 1). Animals treated with test antibody at 0.5 mg / kg had an 80% survival rate, and animals treated with 1.7 mg / kg or greater had a 100% survival rate, while the control group had a 20% survival rate at day 11. The median survival time in the control group was 6.5 days.

[0218] Prophylactic treatment with 0.5 mg / kg or greater of the test antibody resulted in a significant improvement in survival compared to the control group.

[0219] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The body weight per mouse on day 0 was used as the baseline, and body weight change was determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 0.5 mg / kg or higher of the test antibody resulted in a significant reduction in body weight loss compared to the control group (see Figure 8).

[0220] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intraperitoneal administration of the test antibody at 0.5 mg / kg or higher provided a statistically significant improvement in survival and reduced weight loss compared to the control group, whereas 8 of 10 animals in the control group did not survive.

[0221] Example 2 CV3-25 The objective of this study was to evaluate the pre-exposure efficacy of monoclonal antibody CV3-25 after intranasal administration in the SARS-CoV-2 delta K18 hACE2 Tg mouse model.

[0222] On day -1 of the study, animals (n=10 per group) were treated with a range of intranasal doses of test CV3-25 (15-0.2 mg / kg) or PBS control as vehicle. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0223] Materials and Methods The monoclonal antibody CV3-25 was dissolved and diluted in phosphate buffered saline (PBS) to prepare a CV3-25 dilution.

[0224] Dilutions of CV3-25 were made at various concentrations so that administration of 50 μL resulted in doses of CV3-25 ranging from 15 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg relative to the mean body weight of the dosing group.

[0225] The formulations were stored at -80°C ± 10°C and the temperature of the storage units was monitored.

[0226] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0227] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ Stock Number: 034860) mouse model, weighing approximately 17-26 g on day -9. Sixty mice, 11-13 weeks old on the day of test material administration, were used, with 83.3% female. Ten animals were randomly assigned to six treatment groups based on their body weight on day -9 to create groups with similar average body weights. Males comprised 20% of the animals per group, except for the 15 mg / kg group, which was all female. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets and acidified water for animal nutrition, and red plastic tunnels. Each cage system housed 3-5 mice per cage. All mice were moved to new cages every 14 days. Food and water were checked daily and replenished as needed.

[0228] Study design The dose levels of CV3-25 administered in this example were extrapolated from the dose range proven to have prophylactic activity intravenously. Upon arrival, a total of 60 mice, aged 11-13 weeks, were transported to the animal facility and assigned to six experimental groups according to Table 2 (see below). Mice were allowed more than 3 days for acclimatization.

[0229] [Table 2]

[0230] Female SARS-CoV-2 Delta K18 hACE2 mice (20% males per group, except for the 15 mg / kg group, which was all female) were treated via the intranasal route with antibody at doses ranging from 15 mg / kg to 0.2 mg / kg, based on the mean group weight on Day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on Day -1). On Day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for weight loss and mortality until the study was terminated on day 11.

[0231] Antibody administration Upon arrival, the test antibodies were stored at -80°C ± 10°C. The appropriate dose according to the treatment schedule (Table 2) was prescribed according to the mean group weight on day -9, adjusted for a mean weight gain of 4% (based on the subset of animals weighed on day -1).

[0232] Immediately prior to dosing, the material was loaded into a syringe, briefly warmed to room temperature, and then administered to each mouse. Mice were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. The mouse was then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure compound delivery to the lower respiratory tract, the mouse was held for an additional 5-10 seconds before being returned to the nest box.

[0233] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in cold PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Approximately 10 mL of the material was added to each animal using a pipette tip. 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0234] Laboratory analysis The inoculum was returned to the laboratory and replicate samples were titrated on Vero cells to verify the administered virus dose.

[0235] Clinical monitoring General health observations were performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal was weighed daily, beginning one day before infection (day -1).

[0236] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 11. No complete necropsy was performed.

[0237] Data analysis and statistical methods Two animals in the CV3-25 15 mg / kg group failed to dosing (lost part of their antibody volume by sneezing), and these two animals were excluded from statistical analysis.

[0238] Survival fraction, survival time, and body weight change (area under the curve) at day 11 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0239] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0240] survival rate Prophylactic treatment with test antibody at 1.7 mg / kg or greater provided a statistically significant increase in survival compared to the control group (see Figure 2). The median survival time in the control group was 6.0 days.

[0241] Prophylactic treatment with test antibody at 1.7 mg / kg or greater resulted in a significant improvement in survival compared to the control group.

[0242] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The body weight per mouse on day 0 was used as the baseline, and body weight change was determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 0.2 g / kg or higher of the test antibody resulted in a significant reduction in body weight loss compared to the control group (see Figure 9).

[0243] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of the test antibody at doses of 1.7 mg / kg or higher provided a statistically significant improvement in survival and reduced weight loss compared to controls, which exhibited a 10% survival rate and a median survival time of 6 days.

[0244] Example 3 Antibodies according to the present invention The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574 after systemic intraperitoneal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0245] On day -1 of the study, animals (n=10 per group) were treated intraperitoneally with a range of doses of the test antibodies listed above (10-0.2 mg / kg) or a PBS control as vehicle. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0246] Materials and Methods The monoclonal test antibodies described above were buffered with phosphate buffered saline (PBS) and diluted to a final concentration for administration (10-0.2 mg / kg in 200 μL), thereby preparing the test antibody dilutions.

[0247] Test antibody dilutions were made at various concentrations such that administration of 200 μL relative to the average body weight per cage for the dosing group resulted in test antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0248] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0249] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0250] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ Stock Number: 034860) mouse model, weighing approximately 17–22 g on day −5. Sixty mice, 7–11 weeks old, all female, were used. Ten animals were assigned to six treatment groups based on their weight on day −5 to create groups with similar average body weights. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets and acidified water for animal nutrition, and a red plastic tunnel. Each cage system housed 3–5 mice per cage. All mice were transferred to new cages every 14 days. Food and water were inspected daily and replenished as needed. Each animal was weighed daily beginning on the day of infection (day 0).

[0251] Study design The dose levels of the test antibodies applied in this example were based on the maximum possible dosage for the stock concentration of the material (4 mg / mL), as well as on literature and previous experience with an anti-S2 antibody (CV3-25) that demonstrated protective IP at doses of 0.5 mg / kg or higher. A total of 60 mice, 7-11 weeks old, were assigned to six experimental groups according to Table 3 (see below). Mice were allowed more than 3 days for acclimatization.

[0252] [Table 3]

[0253] Female SARS-CoV-2 Delta K18 hACE2 mice were treated via intraperitoneal route with antibody at doses ranging from 10 mg / kg to 0.2 mg / kg, based on the average group weight on day -5. On day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for weight loss and mortality until the study was terminated on day 14.

[0254] Antibody administration Upon arrival, the test antibodies were stored at -80° C.±10° C. Appropriate doses according to the treatment schedule (Table 3) were formulated using the average body weight per cage.

[0255] Prior to dosing, the material was loaded into a 1 mL syringe with a 26G needle and then administered to each mouse. Mice in the treatment group received the indicated dose via intraperitoneal administration of 200 μL of antibody solution into the peritoneal cavity. For administration (200 μL per mouse), mice were grasped by the neck and held upside down at a -45° angle. Mice in the vehicle group, which received PBS, were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. Mice were then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure delivery of the compound to the lower respiratory tract, mice were held for an additional 5–10 seconds before being returned to the nest box.

[0256] Virus administration The virus material was stored at -80°C ± 10°C and kept at room temperature for 10 minutes before administration. 6.3 TCID 50 / mL. Upon thawing, the titer was approximately 10 3.5 TCID 50 The material was diluted in sterile PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0257] Laboratory analysis The actual dose of virus administered was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0258] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0259] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0260] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0261] Viability - i.p. Prophylactic treatment with test antibody at 1.7 mg / kg or higher provides a statistically significant increase in survival compared to the control group (see Figure 3). Animals treated with test antibody at 1.7 mg / kg had a 100% survival rate, 70% at 5 mg / kg, and 90% at 10 mg / kg, while the control group had a 0% survival rate at day 6. The median survival time for the control group was 6 days.

[0262] Prophylactic treatment with 0.2 mg / kg or greater of the test antibody resulted in a significant improvement in survival compared to the control group.

[0263] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed weight was carried forward. The weight per mouse on day 0 was used as the baseline, and body weight changes were determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 0.5 mg / kg or higher of the test antibody resulted in a significant reduction in weight loss compared to the control group.

[0264] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intraperitoneal administration of test antibody at 1.7 mg / kg or higher provided significantly improved survival and reduced weight loss compared to controls, while none of the animals in the control group survived.

[0265] Example 4 Antibodies according to the present invention (intraperitoneal administration test (IP)) The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574 after systemic intraperitoneal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0266] On day -1 of the study, animals (n=10 per group) were treated intraperitoneally with a range of doses of the test antibodies listed above (1.7-0.06 mg / kg) or phosphate buffered saline (PBS) control as a vehicle control. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0267] Materials and Methods Test antibody dilutions were prepared by diluting the above monoclonal test antibodies at 20.41 mg / ml in 20 mM NaAC, 75 mM NaCl, 5% sucrose, pH 5 to a final concentration for administration (1.7 - 0.06 mg / kg in 100 μL).

[0268] Test antibody dilutions were made at various concentrations such that administration of 100 μL for an assumed average body weight of 20 g per mouse resulted in test antibody doses ranging from 1.7 mg / kg, 0.6 mg / kg, 0.2 mg / kg to 0.06 mg / kg.

[0269] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0270] The virus strain tested was the SARS-CoV-2 delta variant (hCOV-19 / Lelystad / B1.617.2 / PW / 2021, lineage AY.5), which had been passaged once in VERO-TMPRSS2.

[0271] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg(B6.Cg-Tg(K18-ACE2)2Prlmn / J (IMSR_JAX:034860) mouse model, weighing approximately 18-20 g. Fifty 7-week-old mice were used, all female. Ten animals were assigned to four treatment groups to create groups with similar average body weights.

[0272] Mice were provided with a shelter for hiding places and nesting material to build nests for sleeping and temperature control. Commercially available food pellets and drinking water were available ad libitum. Water was tested annually for contaminants. Standard laboratory animal feed, RMH-B (Altromin, Germany), was provided. The temperature and relative humidity of the animal room were continuously monitored and recorded daily. The room was maintained at negative pressure, and both inlet and outlet air was HEPA filtered.

[0273] Each animal was weighed daily beginning on the day of infection (day 0).

[0274] Study design The dose levels of the test antibodies applied in this study were based on dose ranges from previous experience. A total of 50 7-week-old mice were assigned to five experimental groups according to Table 4. Mice were allowed more than 3 days for acclimatization.

[0275] [Table 4]

[0276] Female SARS-CoV-2 delta K18 hACE2 mice were treated with antibody at doses ranging from 1.7 mg / kg to 0.06 mg / kg via intraperitoneal administration. On day 0, all mice received a lethal dose (10 per mouse). 3.5 TCID 50 ) and monitored for weight loss and mortality until the study was terminated on day 14.

[0277] Antibody administration Upon arrival, the test antibodies were stored at −80° C.±10° C. Appropriate doses according to the treatment schedule (Table 4) were prescribed based on an average body weight of 20 g per mouse.

[0278] Mice in the treatment and vehicle control groups received the indicated doses by intraperitoneal administration of 100 μL of antibody solution into the peritoneal cavity.

[0279] Virus administration Virus material was stored at -80°C ± 10°C and maintained at a constant constant of 5.6 log per mL before administration. 10 TCID 50 Upon thawing, the titer was approximately 10 per 50 μL. 3.5 TCID 50 The material was diluted with sterile PBS equivalent to 100 mL. Animals were anesthetized with 0.2 mL of a ketamine / xylazine mixture as needed. Each animal received approximately 50 μL (25 μL in each nostril) of the virus dilution. Unused material was frozen at -20°C to -80°C for back titration.

[0280] Laboratory analysis The actual dose of virus administered was verified by back titrating the inoculum and titrating replicate samples on Vero E6 cells.

[0281] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0282] Data analysis and statistical methods Survival rates, survival times, and body weight changes (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted using a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0283] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0284] Viability - i.p. The vehicle control group showed 0% survival and a median survival time of 6 days. Intraperitoneal administration of the test antibody at 1.7 mg / kg provided a statistically significant increase in survival compared to the control group, with 100% survival at this dose (see Figure 4A).

[0285] Prophylactic treatment with 1.7 mg / kg of test antibody resulted in a significant increase in survival compared to the control group.

[0286] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed weight was carried forward. The weight per mouse on day 0 was used as the baseline, and body weight change was determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 1.7 mg / kg of test antibody was required for a significant reduction in weight loss compared to the control group (Figure 4B).

[0287] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intraperitoneal administration of 1.7 mg / kg of the test antibody significantly improved survival and reduced weight loss compared to controls, while none of the animals in the control group survived (see Figure 4A).

[0288] Example 5 Antibodies according to the present invention The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574 after intranasal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0289] On day -1 of the study, animals (n=10 per group) were treated with a range of intranasal doses of the test antibodies described above (10-0.2 mg / kg) or a PBS control. On day 0 of the study, animals were treated with 10 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0290] Materials and Methods The monoclonal antibodies according to the invention were buffered in phosphate buffered saline (PBS) and diluted to a final concentration for administration (10-0.2 mg / kg in 50 μL).

[0291] Monoclonal dilutions were made at various concentrations such that administration of 50 μL relative to the average body weight per cage for the dosing group resulted in test antibody doses ranging from 10 mg / kg, 5 mg / kg, 1.7 mg / kg, 0.5 mg / kg to 0.2 mg / kg.

[0292] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0293] The virus strain tested was SARS-CoV-2 delta variant (Aus / VIC / 18440 / 2021 (B.1.617.2)), which had been passaged three times in Vero-hSLAM cells.

[0294] animal The animal species used was the SARS-CoV-2 Delta K18 hACE2 Tg mouse model, weighing approximately 16.4–21.8 g on the day of test material administration. Sixty mice, 7–11 weeks old, all female, were used. Ten animals were assigned to six treatment groups based on the creation of groups with similar mean body weights based on day -5 weight. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets, acidified water for animal nutrition, and a red plastic tunnel. Each cage system housed three to five mice per cage. All mice were transferred to new cages every 14 days. Food and water were inspected daily and replenished as needed. Each animal was weighed daily, starting on the day of infection (day 0).

[0295] Study design The dose levels of the monoclonal antibodies applied in this example were based on the maximum possible dosage for the stock concentration of the material (4 mg / mL) and on literature and previous experience with an anti-S2 antibody (CV3-25) that demonstrated protective IN at doses of 1.7 mg / kg or higher. A total of 60 mice, 7-11 weeks old, were transported to the animal facility and assigned to six experimental groups according to Table 5 (see below). Mice were allowed more than 3 days for acclimatization.

[0296] [Table 5]

[0297] Female SARS-CoV-2 DeltaK18 hACE2 mice were treated with antibody at doses ranging from 10 mg / kg to 0.2 mg / kg, based on group mean mouse weight per cage, via the intranasal route. On day 0, all mice received a lethal dose (10 3.5 TCID 50 ) and monitored for mortality and weight loss until the study was terminated on day 14.

[0298] Antibody administration Upon arrival, the test antibodies were stored at −80° C.±10° C. Appropriate doses according to the treatment schedule (Table 5) were prescribed based on the average weight per cage.

[0299] Immediately prior to dosing, the material was drawn into a 1 mL syringe equipped with a 26G needle and then administered to each mouse. Mice were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. The mice were then grasped by the neck and held at a +45° angle. 50 μL of compound or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure compound delivery to the lower respiratory tract, the mice were held for an additional 5-10 seconds before being returned to the nest box.

[0300] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in cold PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus equivalent to 100 μg of IgG was given by intranasal inoculation. Unused material was frozen at -20°C to -80°C for back titration.

[0301] Laboratory analysis The administered virus dose was verified by back titrating the inoculum and titrating replicate samples on Vero cells.

[0302] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0303] Data analysis and statistical methods Survival fraction, survival time, and body weight change (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted according to Bonferroni (for two comparisons with vehicle), following a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0304] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0305] Survival rate – intranasal Prophylactic treatment with 0.2 mg / kg or greater of the test antibody provided statistically significant protection compared to the control group (see Figure 5), whose median survival was 6 days.

[0306] Prophylactic treatment with 0.2 mg / kg or greater of the test antibody resulted in a significant improvement in survival compared to the control group.

[0307] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The body weight per mouse on day 0 was used as the baseline, and body weight changes were determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic treatment with 0.2 mg / kg or higher of the test antibody resulted in a significant reduction in body weight loss compared to the control group.

[0308] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of test antibodies at 0.2 mg / kg or higher significantly improved survival and reduced weight loss compared to controls, in which all animals had died by day 6.

[0309] Example 6 Antibodies according to the invention (intranasal administration (IN)) The aim of this study was to evaluate the pre-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574 after intranasal administration in a SARS-CoV-2 Delta K18 hACE2 Tg mouse model.

[0310] In this study, animals (n=10 per group, except for n=9 at 0.6 mg / kg) were treated with intranasal dose ranges of the test antibodies listed above (1.7-0.06 mg / kg) or PBS control as vehicle on day -1 of the study. 3.5 TCID 50 were loaded with the SARS-CoV-2 delta variant.

[0311] Materials and Methods The monoclonal test antibody described above was buffered at 20.41 mg / ml in 20 mM NaAC, 75 mM NaCl, 5% sucrose, pH 5, and diluted to a final concentration for administration (1.7-0.06 mg / kg in 50 μL), thereby preparing the test antibody dilution.

[0312] Test antibody dilutions were made at various concentrations such that administration of 50 μL for an assumed average body weight of 20 g per mouse resulted in test antibody doses ranging from 1.7 mg / kg, 0.6 mg / kg, 0.2 mg / kg to 0.06 mg / kg.

[0313] Antibodies were stored at -80°C ± 10°C, diluents were stored at 4°C until use, and the temperature of the storage units was monitored.

[0314] The virus strain tested was the SARS-CoV-2 delta variant (hCOV-19 / Lelystad / B1.617.2 / PW / 2021, lineage AY.5), which had been passaged once in VERO-TMPRSS2.

[0315] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg(B6.Cg-Tg(K18-ACE2)2Prlmn / J (IMSR_JAX:034860) mouse model, weighing approximately 18-20 g. Forty-nine 7-week-old mice were used, all female. Ten animals (n = 9 for the 0.6 mg / kg group) were assigned to five treatment groups to create groups with similar average body weights.

[0316] Mice were provided with a shelter for hiding places and nesting material to build nests for sleeping and temperature control. Commercially available food pellets and drinking water were available ad libitum. Water was tested annually for contaminants. Standard laboratory animal feed, RMH-B (Altromin, Germany), was provided. The temperature and relative humidity of the animal room were continuously monitored and recorded daily. The room was maintained at negative pressure, and inhaled and exhaled air was HEPA filtered.

[0317] Each animal was weighed daily beginning on the day of infection (day 0).

[0318] Study design The dose levels of the test antibodies applied were based on previous experience with antibodies according to the invention in the K18-hACE2 mouse model. A total of 49 7-week-old mice were assigned to five experimental groups according to Table 6. Mice were allowed more than 3 days for acclimatization.

[0319] [Table 6]

[0320] On day -1, female SARS-CoV-2 delta K18 hACE2 mice were treated with antibody at doses ranging from 1.7 mg / kg to 0.06 mg / kg via the intranasal route. On day 0, all mice received a lethal dose (10 per mouse). 3.5 TCID 50) and monitored for weight loss and mortality until the study was terminated on day 14.

[0321] Antibody administration Upon arrival, the test antibodies were stored at −80° C.±10° C. Appropriate doses according to the treatment schedule (Table 6) were prescribed based on an average body weight of 20 g per mouse.

[0322] Prior to antibody administration, mice in the treatment and vehicle control groups were anesthetized with 0.2 mL of a ketamine / xylazine mixture. Animals received the indicated dose by intranasally administering 25 μL of antibody solution (50 μL per mouse) into each nostril using a sterile pipette tip.

[0323] Virus administration Virus material was stored at -80°C ± 10°C and maintained at a constant constant of 5.6 log per mL before administration. 10 TCID 50 Upon thawing, the titer was approximately 10 per 50 μL. 3.5 TCID 50 The material was diluted with sterile PBS equivalent to 1000 mg / mL. Animals were anesthetized with 0.2 mL of a ketamine / xylazine mixture as needed. Approximately 50 μL (25 μL in each nostril) of the virus dilution was administered to each animal using a sterile pipette tip. Unused material was frozen at -20°C to -80°C for back titration.

[0324] Laboratory analysis The administered virus dose was verified by back titrating the inoculum and titrating replicate samples on Vero E6 cells.

[0325] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0326] Data analysis and statistical methods Survival rates, survival times, and body weight changes (area under the curve) at day 14 were compared with the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups were compared with the vehicle (PBS solution) control group. P values ​​were adjusted using a stepwise approach (starting with the highest antibody dose and conditionally testing lower doses if the previous step was statistically significant).

[0327] Statistical analysis was performed using R, and statistical significance was set at α = 0.05.

[0328] Survival rate – intranasal For the study, the vehicle control group showed a 0% survival rate and a median survival time of 6 days. Prophylactic intranasal treatment with 0.2 mg / kg or higher test antibody provided a statistically significant increase in survival compared to the control group (see Figure 6A).

[0329] Prophylactic intranasal treatment with test antibody at 0.06 mg / kg or higher resulted in a significant increase in survival compared to the control group (FIG. 6A).

[0330] body weight Body weight change was analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed body weight was carried forward. The body weight per mouse on day 0 was used as the baseline, and body weight change was determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day. Prophylactic intranasal treatment with test antibody at 0.06 mg / kg or higher resulted in a significant reduction in body weight loss compared to the control group (Figure 6B).

[0331] conclusion In this lethal SARS-CoV-2 DeltaK18 hACE2 Tg mouse model, prophylactic intranasal administration of the test antibody at 0.2 mg / kg or higher significantly increased survival and reduced weight loss at 0.06 mg / kg or higher compared to the control group (Figure 6). In contrast, none of the animals in the control group survived.

[0332] Example 7 Antibodies according to the present invention The objective of this study is to evaluate the pre- and post-exposure efficacy of a monoclonal antibody according to the invention having heavy chain CDR1 as SEQ ID NO: 055, heavy chain CDR2 as SEQ ID NO: 137, heavy chain CDR3 as SEQ ID NO: 257, light chain CDR1 as SEQ ID NO: 363, light chain CDR2 as SEQ ID NO: 480, and light chain CDR3 as SEQ ID NO: 574 after intranasal administration in an NHP model in which each subject has been pre-screened for the absence of serum antibodies against the SARS-CoV-2 virus.

[0333] NHP subjects are treated with an intranasal dose of a composition comprising the antibody described above. The fixed, or nominal, dose of antibody administered to each NHP subject is 0.01 mg to 10 mg (e.g., 0.01 to 2.0 mg ("low dose") or 2.0 to 10 mg ("high dose") or a control (e.g., vehicle only) on day -1 of the study.

[0334] Materials and Methods Monoclonal antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from a nominal dose of 0.01 mg to 2.0 mg or 2.0 to 10 mg, administered in 50 μL to 500 μL per nostril.

[0335] NHP subjects assigned to the control group (sodium acetate buffer) will receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 50 µL–500 µL per nostril.

[0336] All NHPs will receive an intranasal challenge with SARS-CoV-2 on day 0.

[0337] Samples will be collected during the study, and sample types will include nasal sampling (e.g., nasal swabs, washes, or scrapes), serum and / or bronchoalveolar lavage (BAL) samples. Sampling periods will encompass the duration of the study within the time frame of Day -1 to the end of the study (e.g., a 7- to 21-day follow-up period).

[0338] Samples are analyzed for viral quantification (RT-PCR and / or TCID) as needed. 50 The antibody is analyzed for antibody titer (by ELISA, HPLC, or similar methods).

[0339] animal NHP subjects are used at a weight of 3-12 kg (e.g., 5 kg per animal) at the start of the study. Four to 12 animals are assigned to each treatment group.

[0340] Study design The dose levels of monoclonal antibodies applied in this example are based on maximum doses and lower doses based on allometric calculations derived from mouse studies.

[0341] [Table 7]

[0342] NHP subjects will receive intranasal administration of a composition containing an antibody at a dose (nominal dose) of 0.01 mg to 2.0 mg or 2.0 mg to 10 mg. On day 0, all NHP subjects will be challenged with a dose of SARS-CoV-2 and will be observed for clinical symptoms (e.g., weight loss and / or temperature) and virological measures until the end of the study (e.g., 7 to 21 days).

[0343] Antibody administration Test antibody or vehicle alone is administered (eg, by pipette or spray) to each nostril according to the treatment schedule (Table 7), using a volume of 50 μL to 500 μL per nostril.

[0344] Virus administration The viral material is thawed prior to administration. Once thawed, the material is diluted and each animal receives 50 μL to 500 μL of viral solution per nostril. NHP subjects are administered 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 5 TCID 50 They are infected with SARS-CoV-2.

[0345] Data analysis and statistical methods Clinical symptoms (eg, weight loss and temperature) and virological measures are used to compare treated and control groups.

[0346] Results - Clinical symptoms Prophylactic treatment with low and high dose antibody administration provides a reduction in clinical symptom measures compared to control groups.

[0347] Results – Virological Measurements Prophylactic treatment with low and high doses of antibody administration provides a reduction in virological measurements compared to the control group, with all control NHPs showing a viral load in samples following SARS-CoV-2 challenge.

[0348] conclusion In this SARS-CoV-2 NHP model, prophylactic intranasal administration of 2.0-10 mg (high dose) and 0.01-2.0 mg (low dose) of a test antibody according to the invention provides a reduction in virological measures and / or a reduction in clinical symptoms compared to control NHPs.

[0349] Example 8 Compositions comprising antibodies according to the present invention The objective of this study is to evaluate the pre- and post-exposure efficacy of a composition comprising a first monoclonal antibody according to the invention ("first antibody") having a heavy chain CDR1 as SEQ ID NO: 055, a heavy chain CDR2 as SEQ ID NO: 137, a heavy chain CDR3 as SEQ ID NO: 257, a light chain CDR1 as SEQ ID NO: 363, a light chain CDR2 as SEQ ID NO: 480, and a light chain CDR3 as SEQ ID NO: 574, and a second monoclonal antibody according to the invention ("second antibody") having a heavy chain CDR1 as SEQ ID NO: 645, a heavy chain CDR2 as SEQ ID NO: 646, a heavy chain CDR3 as SEQ ID NO: 647, a light chain CDR1 as SEQ ID NO: 648, a light chain CDR2 as SEQ ID NO: 649, and a light chain CDR3 as SEQ ID NO: 650, following intranasal administration in an NHP model in which each subject has been pre-screened for the absence of serum antibodies to the SARS-CoV-2 virus.

[0350] NHP subjects are treated with an intranasal dose of a composition containing the above-described antibodies (either a single antibody or a combination of both antibodies). The fixed, or nominal, dose of antibody administered to each NHP subject is 0.003 mg to 5 mg or a control (e.g., vehicle only) on day -1 of the study.

[0351] Materials and Methods Monoclonal antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from a nominal dose of 0.01 mg to 20 mg, administered in 50 μL to 500 μL per nostril.

[0352] NHP subjects assigned to the control group (sodium acetate buffer) will receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 50 µL–500 µL per nostril.

[0353] All NHPs will receive an intranasal challenge with SARS-CoV-2 on day 0.

[0354] Samples will be collected during the study, and sample types will include nasal sampling (e.g., nasal swabs, washes, or scrapes), serum and / or bronchoalveolar lavage (BAL) samples. Sampling periods will encompass the duration of the study within the time frame of Day -1 to the end of the study (e.g., a 7- to 21-day follow-up period).

[0355] Samples are analyzed for viral quantification (RT-PCR and / or TCID) as needed. 50 The antibody is analyzed for antibody titer (by ELISA, HPLC, or similar methods).

[0356] animal NHP subjects are used at a weight of 3-12 kg (e.g., 5 kg per animal) at the start of the study. Four to 12 animals are assigned to each treatment group.

[0357] Study design The dose levels of the monoclonal antibodies and combinations applied in this example are based on maximum dosages and on allometric calculations derived from mouse studies.

[0358] [Table 8]

[0359] NHP subjects will receive intranasal administration of a composition containing any single antibody at a nominal dose of 0.003-5 mg (treatment group 1 or 2). Other NHP subjects will receive intranasal administration of a composition containing a mixture of the first and second antibodies at a nominal dose of 0.003-5 mg (treatment group 3). On day 0, all NHP subjects will be challenged with a dose of SARS-CoV-2 and will be observed for clinical symptoms (e.g., weight loss and / or temperature) and virological measures until the end of the study (e.g., 7-21 days).

[0360] Antibody administration According to the treatment schedule (Table 8), antibody compositions (treatment groups 1-3) or vehicle alone are administered (e.g., by pipette or spray) to each nostril using volumes of 50 μL to 500 μL per nostril.

[0361] Virus administration The viral material is thawed prior to administration. Once thawed, the material is diluted and each animal receives 50 μL to 500 μL of viral solution per nostril. NHP subjects are administered 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 5 TCID 50 They are infected with SARS-CoV-2.

[0362] Data analysis and statistical methods Clinical symptoms (e.g., weight loss and temperature) and virological measures are used to compare treatment groups (e.g., treatment groups 1-3) with the control group and / or between treatment groups (e.g., treatment group 1 versus treatment group 3).

[0363] Results - Clinical symptoms Prophylactic treatment with treatments (shown in Table 8) provides a reduction in clinical symptom measures compared to the control group or between treatment groups.

[0364] Results – Virological Measurements Prophylactic treatment with treatments (shown in Table 8) provides a reduction in virological measures compared to the control group or between treatment groups.

[0365] conclusion In this SARS-CoV-2 NHP model, prophylactic intranasal administration of a treatment according to the invention (shown in Table 8) provides a reduction in clinical symptoms and / or a reduction in virological measures compared to control NHPs and in comparisons between treatments.

[0366] Example 9 Bispecific antibodies according to the present invention The purpose of this intranasal mouse study was to evaluate the pre- and post-exposure efficacy of bispecific monoclonal antibodies according to the invention, which have a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, compared to parental antibodies. The first parent antibody ("Parent mAb 1") has the following CDRs represented by SEQ ID NOs: 055, 137, 257, 363, 480, and 574. The second parent antibody ("Parent mAb 2") has the following CDRs represented by SEQ ID NOs: 645, 646, 647, 648, 649, and 650. Bispecific antibodies according to the invention have a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 645, 646, 647, 648, 649-650.

[0367] Animals are treated on day -1 of the study with intranasal nominal doses (0.001 mg / kg to 15 mg / kg) containing either parental mAb 1 or parental mAb 2, or the bispecific antibodies described above, or vehicle control.

[0368] Materials and Methods The parent monoclonal antibodies and bispecific antibodies according to the invention are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from 0.001 mg / kg to 15 mg / kg, administered in 25 μL to 50 μL per nostril.

[0369] Mice assigned to the control group (sodium acetate buffer) receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 25 µL–50 µL per nostril.

[0370] All mice will receive an intranasal challenge with SARS-CoV-2 delta on day 0.

[0371] animal SARS-CoV-2 Delta K18 hACE2 Tg mice are used at the start of the study, weighing approximately 15 g to 22 g (e.g., 20 g per animal). Six to ten animals are assigned to each treatment group.

[0372] Study design The dose levels of the monoclonal and bispecific antibodies applied in this example are based on maximum dosages and on previous results of efficacy studies in mice using the parental antibody alone. Starting at a maximum dose of 15 mg / kg, a 3- or 4-fold dilution curve is applied for each parental or bispecific antibody.

[0373] [Table 9]

[0374] Mice are treated with compositions containing either the parental or bispecific antibodies at doses ranging from 0.001 to 15 mg / kg via the intranasal route. On day 0, all mice are challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (e.g., 10-15 days).

[0375] Antibody administration According to the treatment schedule (Table 9), the parental monoclonal or bispecific antibody or vehicle alone will be administered (e.g., by pipette or spray) to each nostril using a volume of 25 μL to 50 μL per nostril.

[0376] Virus administration The viral material was thawed prior to administration. Once thawed, the material was diluted and each animal received 25 μL to 50 μL of virus per nostril. Mice were 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 3.5 TCID 50 They are infected with SARS-CoV-2.

[0377] Laboratory analysis The inoculum is returned to the laboratory and replicate samples are titrated on Vero cells to verify the administered virus dose.

[0378] Clinical monitoring General health observations will be performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal will be weighed daily beginning one day before infection (day -1).

[0379] Data analysis and statistical methods Survival fraction, survival time, and weight change (area under the curve) are compared with corresponding control groups using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups are compared to the vehicle control group. P values ​​are adjusted according to Bonferroni (for triplicate comparisons with vehicle), followed by a stepwise approach within the antibody (starting with the highest dose and conditionally testing lower doses if the previous step is statistically significant).

[0380] A survival dose-response curve was fitted for each of the treatments (parental mAb 1, parental mAb 2, and bispecific) and the ED 50 The efficacy of the bispecific antibody is then compared to the parent antibody.

[0381] survival rate Prophylactic treatment with parental mAb 1, parental mAb 2, or the bispecific antibody (shown in Table 9) provided statistically significant protection against mortality compared to the control group and a significant improvement in survival compared to the control group.

[0382] body weight Weight change is analyzed using area under the curve (AUC) analysis. If a mouse dies or is euthanized during the study, the last observed weight is carried forward. The weight per mouse on day 0 is used as the baseline, and weight change is determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day.

[0383] Prophylactic treatment with the parental or bispecific antibodies (shown in Table 9) provides a statistically significant reduction in weight loss compared to the control group.

[0384] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of bispecific antibodies according to the invention (shown in Table 9) provides a significant improvement in survival and reduced weight loss compared to the control group. Combination index results suggest at least comparable, additive, or possibly synergistic effects of the bispecific antibodies compared to the parental antibodies.

[0385] Example 10 Bispecifics according to the present invention The aim of this intranasal mouse study was to evaluate the pre- and post-exposure efficacy of a bispecific monoclonal antibody according to the invention, having a first Fab capable of binding to the stem helix of SARS-CoV-2 and a second Fab capable of binding to the fusion peptide of SARS-CoV-2, compared to the parent antibodies. The first parent antibody ("Parent mAb 1") has the following CDRs represented by SEQ ID NOs: 055, 137, 257, 363, 480, and 574. The second parent antibody ("Parent mAb 2") has the following CDRs represented by SEQ ID NOs: 639, 640, 641, 642, 643, and 644. The bispecific antibody according to the invention has a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, and 644.

[0386] Animals (n=8 per group) were treated on day −1 of the study with intranasal doses including 0.007 mg / kg to 1.7 mg / kg (parental mAb 1 or parental mAb 2), or 0.002 mg / kg to 10 mg / kg (bispecific antibody), as described above, or vehicle control.

[0387] Materials and Methods The parent monoclonal antibodies were provided in sodium acetate buffer (20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and the bispecific antibodies according to the invention were provided in phosphate buffered saline (PBS), with all antibodies diluted to a final concentration for administration ranging from either 0.007 mg / kg to 1.7 mg / kg (parent mAb 1 or parent mAb 2) or 0.002 mg / kg to 10 mg / kg (bispecific antibody) and administered in 25 μL per nostril.

[0388] Mice assigned to the control group received vehicle (phosphate buffered saline, PBS), administered at 25 μL per nostril.

[0389] All mice will receive an intranasal challenge with SARS-CoV-2 delta on day 0.

[0390] animal The animal species used was the SARS-CoV-2 delta K18 hACE2 Tg (B6.Cg-Tg(K18-ACE2)2Prlmn / JArc; Jax™ Stock Number: 034860) mouse model, weighing approximately 13.6-22.6 g on day 0. A total of 144 mice, all female, were used, aged 7-11 weeks. Eight animals were assigned to each of the 17 treatment groups, creating groups with similar average body weights. Eight mice were assigned to the vehicle control group. All mice were housed in individually ventilated cages (IVCs) with corncob bedding, tissue or shredded paper as nesting material, wooden chew blocks, accessible food pellets and acidified water for animal nutrition, and a red plastic tunnel. Each cage system housed 3-5 mice per cage. All mice were moved to new cages every 14 days. Food and water were checked daily and replenished as needed. Each animal was weighed daily beginning on the day of infection (day 0).

[0391] Study design The dose levels of the parental and bispecific antibodies applied in this example were based on previous experience with the parental antibodies, which demonstrated intranasal protection at doses of 0.2 mg / kg or higher (parental mAb 1) and 0.5 mg / kg or higher (parental mAb 2) administered intranasally. The efficacy of the two parental bispecific antibodies was expected to be comparable, and the bispecific antibodies were included at doses one dose step up and down from the parental antibody dose range. A total of 144 mice, aged 7 to 11 weeks, were assigned to 18 experimental groups according to Table 10 (see below). Mice were allowed more than 3 days for acclimatization.

[0392] [Table 10]

[0393] Mice were treated with compositions containing either the parental or bispecific antibodies at doses ranging from 0.002 to 10 mg / kg via the intranasal route. On day 0, all mice were challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (day 14).

[0394] Antibody administration Upon arrival, the test antibodies were stored at -80° C.±10° C. Appropriate doses according to the treatment schedule (Table 10) were prescribed based on the average weight per cage.

[0395] Immediately prior to dosing, materials prepared according to Table 10 were loaded into a 1 mL syringe with a 26G needle and then administered to each mouse. Mice were anesthetized with isoflurane (4% v / v in 2 L / min O2) for 2 minutes. The mice were then grasped by the neck and held at a +45° angle. 50 μL of material or vehicle control was then slowly administered into each mouse's nostril (25 μL in each nostril). To ensure delivery of the compound to the lower respiratory tract, the mice were held for an additional 5–10 seconds before being returned to the nest box.

[0396] Virus administration The virus material was stored at -80°C ± 10°C and thawed before administration. 3.5 TCID 50 The material was diluted in cold PBS equivalent to 50 μL / mL. Animals were anesthetized with isoflurane (4% v / v in 2 L / min O) as needed. Each animal received approximately 10 3.5 TCID 50 Approximately 50 μL (25 μL in each nostril) of virus, equivalent to 100 μL of nasal inoculation, was given by intranasal inoculation.

[0397] Final Inspection Mice were euthanized by cervical dislocation at the end of the study on day 14. No complete necropsy was performed.

[0398] Data analysis and statistical methods Survival fraction, survival time, and body weight change (using area under the curve) at day 14 were compared with the vehicle control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. P values ​​were adjusted according to Bonferroni for the four comparisons to the vehicle control group, followed by a stepwise approach starting with the highest dose within an arm and conditionally testing lower doses if the previous dose was statistically significant.

[0399] Probit regression was used to estimate survival dose-response curves without assuming parallel dose-response curves for treatments using the survival fraction at day 14. The model was used to estimate the median effective dose and to determine the ratio between treatments.

[0400] Statistical analyses were performed using R, and Probit regression was performed using the R package drc (v 3.2.0). Statistical significance was set at α = 0.05.

[0401] survival rate The vehicle control group showed 0% survival and a median survival time of 7 days. Prophylactic intranasal treatment with 0.11 mg / kg or more of either parental antibody intranasally provided a statistically significant increase in survival compared to the control group (see Figures 7A and 7D). Prophylactic treatment with 0.16 mg / kg or more of the bispecific antibody provided a statistically significant increase in survival compared to the control group (Figure 7G).

[0402] Prophylactic intranasal treatment with either parental antibody at 0.03 mg / kg or higher or bispecific antibody at 0.039 mg / kg or higher intranasally provides a statistically significant increase in survival compared to the control group (see Figures 7A, 7D, and 7G).

[0403] Survival dose-response curves Probit regression was used to estimate survival dose-response curves without assuming parallel dose-response curves between treatments. A vehicle control group was included in the model at a dose of 0 mg / kg. Probit regression models were used to estimate the antibody dose corresponding to 50% survival (ED 50) was estimated (see Table 11). The intranasal median effective dose was lowest for the bispecific antibody, at 0.013 mg / kg.

[0404] [Table 11]

[0405] body weight Body weight changes were analyzed using area under the curve (AUC) analysis. If a mouse died or was euthanized during the study, the last observed weight was carried forward. The weight per mouse on day 0 was used as the baseline, and body weight changes were determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day.

[0406] Prophylactic intranasal treatment with ≥0.039 mg / kg of the bispecific antibody (Figure 7H) or ≥0.03 mg / kg of the parental antibody (Figures 7B and 7E) resulted in a significant reduction in weight loss compared to the control group.

[0407] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of a bispecific antibody according to the invention (shown in Table 11 and Figure 7G) provides a significant increase in survival compared to the control group at doses of 0.16 mg / kg and above.

[0408] Example 11 Compositions Comprising Antibody Combinations According to the Invention The purpose of this intranasal mouse study is to evaluate the pre- and post-exposure efficacy of a composition comprising a first antibody of the invention capable of binding to the stem helix of SARS-CoV-2 and a second antibody capable of binding to the fusion peptide of SARS-CoV-2. The first antibody ("first mAb") has a heavy chain CDR1 as SEQ ID NO: 055, a heavy chain CDR2 as SEQ ID NO: 137, a heavy chain CDR3 as SEQ ID NO: 257, a light chain CDR1 as SEQ ID NO: 363, a light chain CDR2 as SEQ ID NO: 480, and a light chain CDR3 as SEQ ID NO: 574. The second antibody ("second mAb") has a heavy chain CDR1 as SEQ ID NO: 645, a heavy chain CDR2 as SEQ ID NO: 646, a heavy chain CDR3 as SEQ ID NO: 647, a light chain CDR1 as SEQ ID NO: 648, a light chain CDR2 as SEQ ID NO: 649, and a light chain CDR3 as SEQ ID NO: 650.

[0409] Animals are treated on day -1 of the study with either an intranasal dose (0.001 mg / kg to 15 mg / kg) containing the first mAb or the second mAb, or a composition containing the first and second antibodies described above, or a vehicle control.

[0410] Materials and Methods The first and second mAbs are provided in sodium acetate buffer (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5) and diluted to a final concentration for administration ranging from 0.001 mg / kg to 20 mg / kg, administered as 25 μL to 50 μL per nostril.

[0411] Mice assigned to the control group (sodium acetate buffer) receive vehicle (e.g., 20 mM sodium acetate, 75 mM sodium chloride, 5% sucrose, pH 5.5), administered as 25 µL–50 µL per nostril.

[0412] All mice will receive an intranasal challenge with SARS-CoV-2 delta on day 0.

[0413] animal SARS-CoV-2 Delta K18 hACE2 Tg mice are used at the start of the study, weighing approximately 15 g to 22 g (e.g., 20 g per animal). Six to ten animals are assigned to each treatment group.

[0414] Study design The dose levels for the first or second mAb, or compositions comprising the first and second antibodies described above, in this example are based on maximum dosages and on previous results of efficacy studies in mice using the parent antibody alone, with a 3- or 4-fold dilution curve applied for each parent antibody or bispecific antibody, starting at a maximum dose of 15 mg / kg or less.

[0415] [Table 12]

[0416] Mice are treated via intranasal administration with a composition containing either the first or second antibody or a composition containing the combination at a dose ranging from 0.001 to 15 mg / kg. On day 0, all mice are challenged with a dose of SARS-CoV-2 and monitored for survival and body weight until the end of the study (e.g., 10-15 days).

[0417] Antibody administration According to the treatment schedule (Table 12), a composition containing the first or second antibody or combination, or vehicle alone, is administered to each nostril (e.g., by pipette or spray) using a volume of 25 μL to 50 μL per nostril.

[0418] Virus administration The viral material was thawed prior to administration. Once thawed, the material was diluted and each animal received 25 μL to 50 μL of viral solution per nostril. Mice were 10 2 ~10 8 TCID 50 of SARS-CoV-2 (e.g., 10 3.5 TCID 50 They are infected with SARS-CoV-2.

[0419] Laboratory analysis The inoculum is returned to the laboratory and replicate samples are titrated on Vero cells to verify the administered virus dose.

[0420] Clinical monitoring General health observations will be performed on each animal at least once daily (during normal work procedures) from the day of arrival until the end of the study. Each animal will be weighed daily beginning one day before infection (day -1).

[0421] Data analysis and statistical methods Survival fraction, survival time, and weight change (area under the curve) are compared to the control group using Fisher's exact test, log-rank, and Welch's t-test, respectively. All groups are compared to the vehicle control group. P values ​​are adjusted according to Bonferroni (for triplicate comparisons with vehicle), followed by a stepwise approach within the antibody (starting with the highest dose and conditionally testing lower doses if the previous step is statistically significant).

[0422] Combination Index A survival dose-response curve was fitted for each of the treatments (compositions containing the first or second antibody or combination) and the ED 50 The effectiveness of the combination is then compared to the first and second antibodies by calculating a combination index.

[0423] survival rate Prophylactic treatment with a composition comprising the first or second antibody or combination (shown in Table 12) compared to the control group provides statistically significant protection against mortality and significant improvement in survival compared to the control group.

[0424] body weight Weight change is analyzed using area under the curve (AUC) analysis. If a mouse dies or is euthanized during the study, the last observed weight is carried forward. The weight per mouse on day 0 is used as the baseline, and weight change is determined relative to the baseline, with the net AUC defined as the sum of the areas above and below the baseline using the percentage change per day.

[0425] Prophylactic treatment with a composition containing the combination (shown in Table 12) provides a statistically significant reduction in weight loss compared to the control group.

[0426] conclusion In this SARS-CoV-2 delta mouse model, prophylactic intranasal administration of a composition comprising a combination according to the invention (shown in Table 12) provides a significant improvement in survival and reduced weight loss compared to the control group. The combination index results suggest at least comparable, additive, or possibly synergistic effects for the combination compared to the first or second antibody administered alone.

[0427] Example 12 ELISA binding assessment of antibodies according to the invention The purpose of this study was to evaluate antibody binding to various coronaspike antigens of various alpha- and beta-coronaviruses.

[0428] Binding was tested by enzyme-linked immunosorbent assay (ELISA). Briefly, spike antigens were used to coat the surface of microwells of an ELISA plate. Decreasing concentrations of antibodies were then added in duplicate to the coated, pre-blocked microwell surface, allowing them to attach to the coated antigen. Detection antibodies were added, labeled with horseradish peroxidase (HRP), and a colorimetric signal was generated upon addition of substrate. The readout was optical density, reflecting binding. Antibodies were assayed in duplicate against several alpha- and beta-coronavirus spike antigens, including, but not limited to, SARS-CoV-2 delta, Omicron XBB.1.5, and human coronavirus NL63. Antibodies were tested in a range of 10.0–0.000002 nM relative to other anti-S1 antibodies. The half-maximal effective concentration (EC) was used for each antibody on each antigen as an estimate of binding affinity. 50 ) was calculated.

[0429] [Table 13]

[0430] ELISA binding assays (Table 13) showed that the antibodies according to the invention were able to effectively bind to all spike antigens tested from SARS-CoV-2 delta and betacoronaviruses, including Omicron XBB.1.5. The alphacoronavirus NL63 did not bind to the antibodies. The anti-S1 antibody tested bound only to SARS-CoV-2 delta and showed no breadth among the alphacoronaviruses NL63, as expected from the literature. The isotype control antibody did not show any binding to coronavirus antigens.

[0431] Example 13 Affinity binding evaluation of the antibody according to the present invention by MSD The goal of this study was to evaluate the relative binding affinity for spike antigens in a multiplexed method with extremely high specificity and low sample input. Therefore, antibodies were tested in Mesoscale Discovery (MSD), a multiplexed assay using electrochemiluminescent labels conjugated to detection antibodies. In this assay, up to 10 trimeric viral antigens are individually printed onto spots in each well of a plate, allowing for multiplexed assay readout. Antibodies according to the present invention bind to the viral antigens through Fab-mediated recognition, followed by the addition of an anti-human IgG sulfotag detection antibody, which recognizes the human IgG Fab. Addition of a read buffer containing substrate and passage of current through the plate electrodes initiates an electrochemical (current), buffer-substrate, and luminescent (light) cascade, resulting in light emission. The intensity of the emitted light is measured for each spot, revealing the level of analyte binding. Antibodies were assayed in duplicate in separate assays against spike antigens of SARS-CoV-2, SARS-CoV-1, MERS, hCoV NL63, hCoV HKU, hCoV OC43, and hCoV 229E (V-Plex COVID-19 Coronavirus Panel 3 (IgG) Kit cat# K15399U-2) (Table 15), as well as a set of Omicron variants (V-Plex SARS-CoV-2 Panel 34 (IgG) Kit cat# K15690U-2) (Table 15). Antibodies were tested at a dilution range starting at 555 ng / ml for Panel 3 and 10 ng / ml for the Omicron Panel 24 Kit. Light emission from the MSD sulfo-tag antibodies was quantified on an MSD Discovery WorkBench. Using the calibration curve, antibody concentrations were calculated by fitting raw electrochemiluminescence unit (eCLU) data from the calibrators to a logistic regression for curve fitting (sigmoidal, 4PL, X = concentration). Model asymptote was constrained to the lower limit of detection (LLOQ) and upper limit of detection (ULOQ). The concentration (pg / ml) corresponding to the midpoint of the dynamic range (i.e., the range between the lower and upper limits of quantitation (LLOQ and ULOQ)) was reported.

[0432] [Table 14]

[0433] MSD binding arrays (see Table 14) demonstrated that the antibodies described herein effectively bind to all spike antigens from betacoronaviruses, including SARS-CoV-2, MERS, HKU1, and OC43. The alphacoronavirus NL63 did not bind to the antibodies, while the alphacoronavirus 229E bound at high concentrations. Other hCoV betacoronaviruses, HKU1 and OC43, effectively bound the antibodies described herein, demonstrating breadth among betacoronaviruses, including MERS and the seasonal cold coronaviruses OC43 and HKU1, and limited breadth among alphacoronaviruses for 229E. The SARS-CoV-2 receptor-binding domain (RBD) in the S1 portion of the spike bound to the antibodies described herein only at very high concentrations. The anti-S1 antibodies tested here bound to the RBD domain and wild-type SARS-CoV-2 at low concentrations, as expected from the literature, and did not demonstrate breadth among other coronaviruses. An isotype control antibody showed no binding to any coronavirus antigens.

[0434] [Table 15]

[0435] The MSD binding array showed that the antibodies of the present invention were able to effectively bind to all omicron variants tested in this assay, even at concentrations lower than Wuhan (Table 15). This is consistent with the results of omicron XBB.1.5 binding in the ELISA assay (Table 14). The BA.1 and BN.1 variants of SARS-CoV-2 bound particularly at low concentrations, indicating that the antibodies of the present invention retained binding efficacy against all omicron variants tested. The anti-S1 control antibody bound to the SARS-CoV-2 spike at lower concentrations than the antibodies of the present invention, but bound early omicron variants only at very high concentrations and did not bind later omicron variants such as BQ.1 and XBB.1. The isotype control antibody showed no binding to coronavirus antigens at all.

[0436] Example 14 Live virus neutralization of antibodies according to the invention The purpose of this study was to evaluate the ability of antibodies to neutralize live coronaviruses. Antibodies were tested for functional activity in live virus microneutralization assays against MERS, SARS-CoV-1, and SARS-CoV-2 Wuhan. Briefly, serial dilutions of antibodies were pre-incubated with the corresponding viruses and then added in quadruplicate to the respective cell lines (Vero: MERS and SARS-CoV-1, SARS-CoV-2: Vero E6 cells). After incubation, cells were fixed and stained with anti-nucleocapsid antibodies, and enzyme-tagged detection antibodies were added. The colored precipitate, which signals the presence of nucleocapsid, was read on an Immunospot analyzer and the 50% inhibitory concentration (IC) was determined by the Zielinska method (REF: https: / / doi.org / 10.1186 / 1743-422X-2-84). 50 ) reported. Antibodies were tested for neutralizing activity in the range of 0.025 to 500 μg / ml.

[0437] [Table 16]

[0438] Overall, the antibodies were able to neutralize the tested viruses SARS-CoV-1, SARS-CoV-2, and MERS at various concentrations (see Table 16), with IC for MERS-CoV. 50 The concentrations were minimal, thus confirming that the antibodies according to the invention are able to neutralize these various betacoronaviruses, as suggested by the binding assessment by MSD (Table 15a). The positive serum control for SARS-CoV-2 showed a high MN 50 The titer is shown.

[0439] Example 15 Neutralization of pseudovirions by antibodies of the present invention The purpose of this study was to evaluate the ability of antibodies to neutralize various pseudotyped virus particles. Advantages over live virus assays include the ability to use pseudotyped viruses in laboratories with lower biosafety levels, the ability to produce and test novel variants of concern more rapidly, and broader testing across variants of concern or interest. Briefly, pseudovirions were produced by cotransfecting HEK293T cells with a virus-expressing plasmid and the pHIV-1NL43 ΔEnv-NanoLuc reporter virus plasmid. A dilution series of antibodies was then preincubated with the corresponding pseudovirus and added to HEK293T cells expressing ACE2, the entry receptor for SARS-CoV-2, SARS-CoV-1, and NL63. For hCoV 229E, diluted antibodies mixed with the virus were added to Huh7 cells expressing the aminopeptidase N (APN) receptor, which facilitates hCoV 229E entry into host cells. After incubation, cells were washed and lysed, and luciferase activity in the cell lysates was measured by reading relative optical units (RLU) using the Nano-Glo Luciferase Assay System and the GloMax System. The 50% inhibitory concentration (IC) was determined as the antibody concentration at which infectivity was inhibited by 50% using a four-parameter logistic regression (4PL) curve fit.50 The bispecific antibodies were tested for neutralizing activity in the range of 0.0042 to 250 μg / ml against SARS-CoV-1, SARS-CoV-2 Wuhan, Delta, Omicron BA4, Omicron BQ.1.1, Omicron XBB.1, hCoV NL63, and hCoV 229E.

[0440] [Table 17]

[0441] Overall, the antibodies were able to neutralize the tested betaviruses SARS-CoV-1 and SARS-CoV-2, including variants of concern, at various concentrations (see Table 17), with an overall trend of higher IC for the Omicron variants. 50 As expected based on the lack of binding in the MSD assay for NL63, the antibody did not neutralize the alphacoronaviruses hCoV NL63 and 229E. Although 229E bound in the MSD assay (Table 16a), the activity did not translate into detectable neutralizing activity in this pseudotype neutralization assay.

[0442] Example 16 Epitope mapping of the antibody according to the present invention The goal of this study was to precisely determine antibody-bound amino acid sequences in spike proteins across various alpha- and beta-coronaviruses. The method involves mapping linear epitopes using a library of overlapping synthetic peptides derived from spike proteins. Briefly, linear epitopes of sequences of interest were synthesized directly on a solid-state chip, generating a library of linear mimetics to aid in the identification of the correct amino acid sequence for target antibodies. The peptide library consisted of overlapping 15-amino acid fragments, with a 14-amino acid overlap. Antibody binding to each of the synthesized peptide chips was tested by enzyme-linked immunosorbent assay (ELISA) assay via incubation of the peptide array with an antibody solution. After washing, the peptide array was incubated with an antibody-peroxidase conjugate, substrate was added, and the reaction was stopped with hydrogen peroxide. The color development was measured and the intensity was reported.

[0443] The antibodies bound to peptide arrays of alpha- and beta-coronaviruses in specific epitopes of 10-20 amino acids (see Figure 16). This method allowed for high confidence in the definition of the epitopes, although there was some variability in the outer regions of the epitopes for each strain, indicated by asterisks.

[0444] Example 17 Affinity binding assessment of bispecific antibodies by MSD The purpose of this study was to evaluate the relative binding affinities to spike antigens in a multiplexed method with extremely high specificity and low sample input. Therefore, bispecific antibodies were tested in comparison to the parent antibodies in Mesoscale Discovery (MSD), a multiplexed assay that uses electrochemiluminescent labels conjugated to the detection antibodies.

[0445] The first parent antibody ("Parent mAb 1") has the following CDRs set forth in SEQ ID NOs: 055, 137, 257, 363, 480, and 574. The second parent antibody ("Parent mAb 2") has the following CDRs set forth in SEQ ID NOs: 639, 640, 641, 642, 643, and 644. A bispecific antibody according to the invention has a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, and 644.

[0446] Using the same production technology, which allows for slightly varying formats, three versions of the bispecific antibody were produced and all tested with the parental mAb 1 and parental mAb 2 combination.

[0447] Bispecific 1 (filled circle) (according to the present invention) is also described inter alia in section 26 and sections 29 and 30, and has a first arm heavy chain amino acid sequence set forth in SEQ ID NO: 663 and a light chain amino acid sequence set forth in SEQ ID NO: 664, and a second arm having a heavy chain amino acid sequence set forth in SEQ ID NO: 665 and a light chain amino acid sequence set forth in SEQ ID NO: 666.

[0448] Bispecific 2 (open diamond) (according to the present invention) is also described inter alia in section 26 and sections 31 and 32 and has a first arm heavy chain amino acid sequence set forth in SEQ ID NO: 667 and a light chain amino acid sequence set forth in SEQ ID NO: 668, and a second arm having a heavy chain amino acid sequence set forth in SEQ ID NO: 669 and a light chain amino acid sequence set forth in SEQ ID NO: 670.

[0449] Bispecific 3 (open box) (according to the present invention) is also described inter alia in paragraphs 26 and 33 and has a first arm heavy chain amino acid sequence set forth in SEQ ID NO: 667 and a light chain amino acid sequence set forth in SEQ ID NO: 668, and a second arm having a heavy chain amino acid sequence set forth in SEQ ID NO: 671 and a light chain amino acid sequence set forth in SEQ ID NO: 672.

[0450] In this assay, up to 10 trimeric viral antigens are individually printed onto spots in each well of the plate, allowing for a multiplexed assay readout. The parent and bispecific antibodies bind to the viral antigens through Fab-mediated recognition, followed by the addition of an anti-human IgG sulfotag detection antibody, which recognizes the human IgG Fab. Addition of a read buffer containing substrate and passage of current through the plate electrodes initiates an electrochemical (current), chemiluminescent (buffer substrate), and luminescent (light) cascade, resulting in light emission. The intensity of the emitted light is measured for each spot, revealing the level of analyte binding. Antibodies were assayed in duplicate against spike antigens from SARS-CoV-2, SARS-CoV-1, MERS, hCoV NL63, hCoV HKU, hCoV OC43, and hCoV 229E (V-Plex COVID-19 Coronavirus Panel 3 (IgG) Kit cat# K15399U-2), as well as a set of Omicron variants (V-Plex SARS-CoV-2 Panel 34 (IgG) Kit cat# K15690U-2) in separate assays. Parental and bispecific antibodies were tested in a dilution curve ranging from 0.25 to 555 ng / ml. The dilution range was adapted to include a wide range, taking into account the potential for reduced potency of bispecific antibodies. Luminescence from the MSD sulfo-tag antibodies was quantified on an MSD Discovery WorkBench. Using the calibration curve, antibody concentrations were back-calculated by fitting raw electrochemiluminescence unit (eCLU) data from the calibrators to a logistic regression for curve fitting (sigmoidal, 4PL, X = concentration). Model asymptote was constrained to the lower limit of detection (LLOQ) and upper limit of detection (ULOQ). The concentration (ng / ml) corresponding to the midpoint of the dynamic range (i.e., the range between the lower and upper limits of quantitation (LLOQ and ULOQ)) was reported.

[0451] [Table 18]

[0452] MSD binding arrays (see Table 18) showed that the bispecific antibodies can effectively bind to all spike antigens from alpha- and beta-coronaviruses. Table 18 reports the binding evaluation of parental mAb 1, parental mAb 2, the three constructed bispecific antibodies (according to the present invention), and the anti-S1 monoclonal antibody against various representative viruses from the α-coronavirus and β-coronavirus genera. These data are presented in Figure 17A.

[0453] The alphacoronavirus NL63 did not bind to the parental anti-stem helix antibody (parental mAb 1) and bound only to the anti-fusion peptide antibody and bispecific antibody, albeit at slightly higher concentrations than the parental antibody. The alphacoronavirus 229E bound to parental mAb 1 at high concentrations and to parental mAb 2 at low concentrations. Other hCoV betacoronaviruses, HKU1 and OC43, bound effectively to parental mAb 1 and also to parental mAb 2 at high concentrations. In contrast, the bispecific antibody bound at intermediate concentrations, suggesting the availability of the Fab fragment of parental mAb 1. The SARS-CoV-2 spike RBD antigen bound to parental mAb 1 (stem helix mAb) and the bispecific antibody only at high concentrations. The control anti-S1 mAb bound only to the SARS-CoV-2 spike and RBD at low concentrations. The data suggest that bispecific antibodies retain the binding capacity derived from each parent antibody, as demonstrated here by utilizing the Fab arm of parent mAb 2 and retaining alphacoronavirus binding activity, particularly for NL63.

[0454] Typically, for bispecific antibodies, binding levels intermediate between those exhibited by parental mAb 1 and parental mAb 2 are expected. Therefore, it is important to note that Bispecific 1, Bispecific 2, and Bispecific 3 (all according to the invention) have acquired broader binding capabilities compared to the parental monoclonal antibodies as far as α- and β-coronaviruses are concerned. Of particular importance is that Bispecific 1, Bispecific 2, and Bispecific 3 have acquired the superior binding properties exhibited by parental mAb 1 with respect to binding to β-coronaviruses, while also acquiring the superior binding properties derived from parental mAb 2 with respect to the α-coronaviruses tested.

[0455] [Table 19]

[0456] MSD binding arrays (see Table 19) showed that the bispecific antibodies could effectively bind to all spike proteins from SARS-CoV-2 omicron variants arranged chronologically from left to right, starting with the original SARS-CoV-2 strain. Only parental mAb 1 retained efficient binding to the later omicron variants, and parental mAb 2 required high antibody concentrations, while the bispecific antibodies retained binding, suggesting that the bispecific antibodies could utilize the Fab arm of parental Fab 1.

[0457] Figure 17B shows the binding capacity of parental mAb 1 (closed triangles) (according to the invention) and parental mAb 2 (open triangles), demonstrating the superior binding capacity of parental mAb 1 to the original SARS-CoV-2 strain and its subsequent SARS-CoV-2 omicron variants.

[0458] Figure 17C retains the parental mAb 1 (closed triangles) (according to the present invention) as a reference point, but instead focuses on the relative binding capacities of the three constructed bispecific antibodies, Bispecific 1 (closed circles), Bispecific 2 (open diamonds), and Bispecific 3 (open squares), against the original SARS-CoV-2 strain and its subsequent SARS-CoV-2 omicron variants.

[0459] Recognizing that Bispecific 2 and Bispecific 3 have broader binding capacities compared to both parental mAbs, while both Bispecific 2 and Bispecific 3 have superior binding capacities for SARS-CoV-2 omicron variants, they do not replace the binding capacity of parental mAb 1.

[0460] Using highly sensitive MSD binding arrays, we surprisingly discovered that Bispecific 1 not only has broader binding capabilities than both parental mAbs, but also consistently exhibits superior binding to classic and recent SARS-CoV-2 omicron variants compared to the parental mAbs as well as Bispecific 2 and Bispecific 3.

[0461] Example 18 Neutralization of live virus by bispecific antibodies The objective of this study was to evaluate the ability of bispecific antibodies to neutralize live coronaviruses and pseudovirions. Bispecific antibodies were tested for functional activity in live virus microneutralization assays against MERS, SARS-CoV-1, SARS-CoV-2 Wuhan, Delta, and Omicron BA.4 / 5.

[0462] The first parent antibody ("Parent mAb 1") has the following CDRs set forth in SEQ ID NOs: 055, 137, 257, 363, 480, and 574. The second parent antibody ("Parent mAb 2") has the following CDRs set forth in SEQ ID NOs: 639, 640, 641, 642, 643, and 644. A bispecific antibody according to the invention comprises a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, and 644.

[0463] The bispecific antibody Bispecific 1 comprises the first Fab and second Fab described above.

[0464] Briefly, serial dilutions of antibodies were pre-incubated with the corresponding viruses and then added to the respective cell lines (Vero: MERS and SARS-CoV-1, all others Vero E6 cells). After incubation, cells were fixed and stained with anti-nucleocapsid antibodies, and enzyme-tagged detection antibodies were added. The colored precipitate, which signals the presence of nucleocapsid, was read on an Immunospot analyzer and the 50% inhibitory concentration (IC 50 ) reported. The bispecific antibodies were tested for neutralizing activity in the range of 0.02 to 500 μg / ml compared to individual control antibodies.

[0465] [Table 20]

[0466] Overall, the bispecific antibodies were able to neutralize the tested viruses SARS-CoV-1, SARS-CoV-2 Wuhan, Delta and Omicron BA.4 / 5, and MERS at various concentrations (see Table 20). The anti-stem helix parental antibodies neutralized SARS-CoV-2 Wuhan and MERS with lower IC compared to the anti-fusion peptide antibodies. 50This trend was also observed for the bispecific antibody, demonstrating that the antibody was able to utilize the anti-stem helix Fab arm, indicating that the bispecific antibody, derived from both Fab arms, retained affinity across the betacoronaviruses tested.

[0467] Example 19 Neutralization of pseudovirions by bispecific antibodies Pseudovirion neutralization assays against pseudotyped virus particles were performed.

[0468] The first parent antibody ("Parent mAb 1") has the following CDRs set forth in SEQ ID NOs: 055, 137, 257, 363, 480, and 574. The second parent antibody ("Parent mAb 2") has the following CDRs set forth in SEQ ID NOs: 639, 640, 641, 642, 643, and 644. A bispecific antibody according to the invention has a first Fab comprising a sequence comprising any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and a second Fab comprising a sequence comprising any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, and 644.

[0469] Bispecific antibody Bispecific 1 comprises the first Fab and second Fab described above.

[0470] The advantage over live virus assays is that pseudotyped viruses can be used in laboratories with low biosafety levels, allowing for more rapid production and testing of novel variants of concern. Briefly, pseudovirions were produced by cotransfecting HEK293T cells with a virus-expressing plasmid and the pHIV-1NL43 ΔEnv-NanoLuc reporter virus plasmid. A dilution series of antibodies was then preincubated with the corresponding pseudovirus and added to HEK293T cells expressing ACE2, the entry receptor for SARS-CoV-2, SARS-CoV-1, and NL63. After incubation, cells were washed and lysed, and luciferase activity in the cell lysates was measured using the Nano-Glo Luciferase Assay System and GloMax System. The 50% inhibitory concentration (IC) was determined as the antibody concentration at which infectivity was inhibited by 50% using a four-parameter logistic regression (4PL) curve fit. 50 The bispecific antibodies were tested for neutralizing activity against SARS-CoV-2 Wuhan, Delta, Omicron BA4 / 5, NQ.1, and XBB.1, hCoV NL63, and hCoV 229E in the range of 0.01 to 250 μg / ml, compared to individual control antibodies.

[0471] [Table 21]

[0472] Pseudovirion neutralization assays (see Table 21) show that parental antibody 1 according to the invention is able to neutralize Omicron variants, especially late-arising variants such as XBB.1.1, at lower concentrations than anti-fusogenic peptide parental antibody mAb 2, with the exception of SARS-CoV-2 Wuhan, which is neutralized by the bispecific antibody at the lowest concentration compared to the parental antibody, and the bispecific antibody has an IC intermediate between the respective parental mAbs. 50 The results show that the bispecific antibody retains neutralizing activity at high concentrations, demonstrating that the bispecific antibody retains the neutralizing affinity derived from each Fab arm of the parent antibody.

[0473] Example 20 Bispecific antibody validation This bispecific binding competition assay demonstrates the ability of bispecific antibodies according to the invention to bind to these two epitopes of coronaviruses, exemplified using the stem helix and fusion peptide found in the SARS-CoV-2 spike protein.

[0474] 1. Antibody production A mammalian cell expression system is used to express and purify a bispecific antibody according to the invention having a first Fab that binds to the stem helix of SARS-CoV-2 and a second Fab that binds to the fusion peptide of SARS-CoV-2, for example, the first Fab has the CDR sequences specified by SEQ ID NOs: 055, 137, 257, 363, 480, and 574, and the second Fab has the CDR sequences specified by SEQ ID NOs: 639, 640, 641, 642, 643, and 644.

[0475] Alternatively, another bispecific antibody according to the invention is expressed and purified, having a first Fab that binds to the stem helix of SARS-CoV-2 and a second Fab that binds to the fusion peptide of SARS-CoV-2, where the first Fab has the CDR sequences specified by SEQ ID NOs: 055, 137, 257, 363, 480, and 574, and the second Fab has the CDR sequences specified by SEQ ID NOs: 645, 646, 647, 648, 649, and 650.

[0476] 2. Characterization of bispecific antibodies The purity and integrity of the bispecific antibodies are confirmed using SDS-PAGE and Western blot analysis. The size and structure of the bispecific antibodies are assessed using techniques such as size exclusion chromatography and mass spectrometry.

[0477] 3. Analysis of Binding Affinity Enzyme-linked immunosorbent assays (ELISAs) are performed to confirm binding to each target antigen separately, i.e., the stem helix of SARS-CoV-2 and then the fusion peptide of SARS-CoV-2.

[0478] Surface plasmon resonance (SPR) or biolayer interferometry (BLI) are used to determine the binding kinetics and affinity of each Fab region to the stem helix of SARS-CoV-2 and then individually to the fusion peptide of SARS-CoV-2. SPR can also be used in competition assays.

[0479] 4. Bispecific Binding Competition Assay for the Stem Helix and Fusion Peptide of SARS-CoV-2 Spike Protein 4.1. Materials and Reagents - a purified bispecific antibody according to the invention (a bispecific antibody targeting both the stem helix of SARS-CoV-2 and the fusion peptide of SARS-CoV-2). - purified monospecific parent antibody 1 having CDR sequences identified by SEQ ID NOs: 055, 137, 257, 363, 480, and 574 and targeting only the stem helix of SARS-CoV-2. - A purified monospecific parent antibody 2 is used, which has the CDR sequences identified by SEQ ID NOs: 639, 640, 641, 642, 643, and 644 and targets only the fusion peptide of SARS-CoV-2. - Alternatively, purified monospecific parent antibody 2 having the CDR sequences identified by SEQ ID NOs: 645, 646, 647, 648, 649, and 650 and targeting only the fusion peptide of SARS-CoV-2. - A purified, labeled, conjugated monospecific parent antibody 1 having the CDR sequences identified by SEQ ID NOs: 055, 137, 257, 363, 480, and 574 and targeting only the stem helices of SARS-CoV-2. - A purified, labeled, conjugated monospecific parent antibody 2 having the CDR sequences identified by SEQ ID NOs: 639, 640, 641, 642, 643, and 644 and targeting only the fusion peptide of SARS-CoV-2. - Alternatively, a purified, labeled, conjugated monospecific parent antibody 2 having the CDR sequences identified by SEQ ID NOs: 645, 646, 647, 648, 649, and 650 and targeting only the fusion peptide of SARS-CoV-2. - Purified SARS-CoV-2 viral spike protein bearing both the SARS-CoV-2 stem-helix epitope and the SARS-CoV-2 fusion peptide epitope. - PBS blocking buffer containing 1% bovine serum albumin (BSA). - PBS washing buffer containing Tween-20 0.05% (v / v). - A secondary direct detection antibody conjugated to an enzyme, or a substrate for an enzyme-conjugated detection antibody. - 96-well microplates and 96-well microplate absorbance or fluorescence intensity readers.

[0480] Procedure Coating of 96-well microplates Pre-assigned wells of a microplate are coated with purified SARS-CoV-2 viral spike protein in coating buffer at a concentration optimized for antigen binding (e.g., optimized using any of 0.1, 0.5, 1.0, 2.0, 3.0, 4.0, 5.0, and 10.0 μg / mL), and the covered microplate is incubated overnight at 4°C.

[0481] Blocking nonspecific binding Wash the SARS-CoV-2 virus spike-coated wells three times with PBS wash buffer. Nonspecific binding is blocked by adding PBS blocking buffer to each well. Incubate at room temperature for 1 hour. Then wash the plate three times with PBS wash buffer.

[0482] Blocking specific binding In the pre-assigned wells, i. PBS blocking buffer and anti-SARS-CoV-2 stem-helix monospecific antibody alone at a concentration optimized to block specific binding; ii. PBS blocking buffer and anti-SARS-CoV-2 fusion peptide monospecific antibody alone at a concentration optimized to block specific binding; iii. PBS blocking buffer and a mixture of anti-SARS-CoV-2 stem-helix monospecific antibodies and anti-SARS-CoV-2 fusion peptide monospecific antibodies at concentrations optimized to block specific binding. Specific binding is blocked by adding HCl and the plate containing these wells is incubated for 1 hour at room temperature.

[0483] Binding competition assay Add the following solutions to pre-assigned wells: Competitor solution 1: Prepare a solution of labeled conjugated monospecific antibodies against the SARS-CoV-2 stem helix in PBS blocking buffer at a concentration optimized for the binding assay. Add competitor solution 1 to the pre-assigned coated wells above and incubate for 2 hours at room temperature. Competitor solution 2: Prepare a solution of labeled conjugated monospecific antibodies against SARS-CoV-2 fusion peptide in PBS blocking buffer at a concentration optimized for the binding assay. Add competitor solution 2 to the pre-assigned coated wells and incubate for 2 hours at room temperature. Competitor solution 3: Prepare a solution of labeled conjugated bispecific antibodies against the SARS-CoV-2 stem helix and SARS-CoV-2 fusion peptide in blocking buffer at a concentration optimized for the binding assay. Add competitor solution 3 to the pre-assigned coated wells and incubate for 2 hours at room temperature.

[0484] Note that assay optimization is required for antibody concentration, incubation time, and other parameters. Ensure controls are included to validate specificity of binding. Assays should be performed in triplicate or as needed for statistical significance.

[0485] 5. Detection Wash the 96-well microplate containing the assays mentioned above three times with PBS wash buffer. If a secondary antibody system is used, add the appropriate detection antibody to each well and incubate for 1 hour at room temperature. If an enzyme-conjugated detection antibody is used, add the substrate according to the manufacturer's instructions.

[0486] 6. Measurement The absorbance or fluorescence intensity of each well is measured using a plate reader. The data is analyzed to compare the signals from wells containing bispecific and monospecific antibodies.

[0487] 7. Data Analysis Calculate the competition between bispecific and monospecific antibodies for binding to the spike protein. Determine the relative binding affinities of bispecific antibodies to epitopes in the SARS-CoV-2 stem helix and SARS-CoV-2 fusion peptide.

[0488] 8. Conclusion The combination of in vitro binding assays, structural analysis, and in vivo validation provides comprehensive evidence supporting the bispecific nature of the antibody and its ability to bind two different target antigens via two distinct Fab regions.

Claims

1. An anti-coronavirus, preferably anti-SARS-CoV-2 bispecific antibody or antigen-binding fragment thereof, having a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, or 574, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 639, 640, 641, 642, 643-644.

2. 2. The bispecific antibody or antigen-binding fragment thereof of claim 1, comprising a first Fab capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, wherein the first Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 055, 137, 257, 363, 480, and / or 574, and the second Fab comprises a sequence comprising preferably any one or more of SEQ ID NOs: 639, 640, 641, 642, 643, and / or 644.

3. a first Fab that specifically binds to the stem helix of a coronavirus, preferably SARS-CoV-2, and a second Fab that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2; a.) a first Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 055, the heavy chain CDR2 region of SEQ ID NO: 137, and the heavy chain CDR3 region of SEQ ID NO: 257, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 363, the light chain CDR2 region of SEQ ID NO: 480, and the light chain CDR3 region of SEQ ID NO: 574; b.) the second Fab comprises a heavy chain variable region comprising, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641, and a light chain variable region comprising, as CDRs, the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644; 3. The bispecific antibody or antigen-binding fragment thereof according to claim 1 or 2,

4. a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2; and b) a light chain comprising the VL-CL domains and a heavy chain comprising the VH-CH1-CH2-CH3 domains of an antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2; wherein the constant domains CL and CH1 from an antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, are replaced by each other.

4. The bispecific antibody or antigen-binding fragment thereof according to claim 1 .

5. the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 055, a heavy chain CDR2 region of SEQ ID NO: 137, and a heavy chain CDR3 region of SEQ ID NO: 257; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 363, a light chain CDR2 region of SEQ ID NO: 480, and a light chain CDR3 region of SEQ ID NO: 574; The VH domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641; and the VL domain of the antibody or antigen-binding fragment thereof capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises, as CDRs, the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO:

644. The bispecific antibody or antigen-binding fragment thereof according to claim 4.

6. a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the constant domains CL and CH1 from an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 4. The bispecific antibody or antigen-binding fragment thereof according to claim 1 .

7. the VH domain of the antibody or antigen-binding fragment thereof capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a heavy chain CDR1 region of SEQ ID NO: 055, a heavy chain CDR2 region of SEQ ID NO: 137, and a heavy chain CDR3 region of SEQ ID NO: 257; and the VL domain of the antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, comprises, as CDRs, a light chain CDR1 region of SEQ ID NO: 363, a light chain CDR2 region of SEQ ID NO: 480, and a light chain CDR3 region of SEQ ID NO: 574; The VH domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the heavy chain CDR1 region of SEQ ID NO: 639, the heavy chain CDR2 region of SEQ ID NO: 640, and the heavy chain CDR3 region of SEQ ID NO: 641, and the VL domain of the antibody capable of binding to the fusion peptide of coronavirus, preferably SARS-CoV-2, comprises as CDRs the light chain CDR1 region of SEQ ID NO: 642, the light chain CDR2 region of SEQ ID NO: 643, and the light chain CDR3 region of SEQ ID NO: 644; The bispecific antibody or antigen-binding fragment thereof according to claim 6.

8. a) a light chain comprising a VL-CL domain and a heavy chain comprising a VH-CH1-CH2-CH3 domain of an antibody capable of binding to the stem helix of a coronavirus, preferably SARS-CoV-2, and b) a light chain comprising VL-CL domains and a heavy chain comprising VH-CH1-CH2-CH3 domains of an antibody capable of binding to the fusion peptide of a coronavirus, preferably SARS-CoV-2 Including, the domains VL-CL and VH-CH1 from an antibody that specifically binds to the fusion peptide of a coronavirus, preferably SARS-CoV-2, are replaced by each other; 4. The bispecific antibody or antigen-binding fragment thereof according to claim 1 .

9. 9. The bispecific antibody or antigen-binding fragment thereof according to claim 1 , wherein the first Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO:

629.

10. 9. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 8, wherein the second Fab comprises a heavy chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO:

673.

11. 9. The bispecific antibody or antigen-binding fragment thereof according to claim 1 , wherein the first Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO:

630.

12. 9. The bispecific antibody or antigen-binding fragment thereof of claim 1 , wherein the second Fab comprises a light chain variable region having an amino acid sequence that is at least 90% identical, at least 91% identical, at least 92% identical, at least 93% identical, at least 94% identical, at least 95% identical, at least 96% identical, at least 97% identical, at least 98% identical, or at least 99% identical to the amino acid sequence set forth in SEQ ID NO:

674.

13. 9. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 8, wherein the bispecific antibody or antigen-binding fragment thereof is selected from the group consisting of a full-length antibody, a Fab, a modified Fab, a Fab', a modified Fab', a F(ab')2, an Fv, a single domain antibody, a scFv, a scFv-Fc, a bivalent, trivalent, or tetravalent antibody, a Bis-scFv, a diabody, a triabody, a tetrabody, and epitope-binding fragments thereof.

14. 14. A method for the production of a bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, comprising: a) culturing a host cell comprising an expression vector comprising a polynucleotide encoding the bispecific antibody or antibody fragment of any one of claims 1 to 13 under conditions that allow the production of said bispecific antibody or antigen-binding fragment thereof; and b) isolating the bispecific antibody or antigen-binding fragment thereof. A method comprising:

15. 14. A nucleic acid molecule, preferably an isolated nucleic acid molecule, comprising a nucleic acid sequence encoding the bispecific antibody or antigen-binding fragment thereof, or the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13.

16. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13 and claim 15, comprising a nucleic acid sequence encoding the heavy chain variable region and / or the light chain variable region of the bispecific antibody or antigen-binding fragment thereof, The nucleic acid sequence is (i) a first nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 055, or a first nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 055 by at most one or two amino acids; (ii) a second nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 137, or a second nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 137 by at most 1, 2, 3, 4, or 5 amino acids; (iii) a third nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 257, or a third nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 257 by at most 1, 2, or 3 amino acids; (iv) a fourth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 363, or a fourth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 363 by at most 1, 2, 3, or 4 amino acids; (v) a fifth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 480, or an amino acid sequence that differs from SEQ ID NO: 480 by at most one or two amino acids; (vi) a sixth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 574, or an amino acid sequence that differs from SEQ ID NO: 574 by at most 1, 2, 3, or 4 amino acids; (vii) a seventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 639, or an amino acid sequence that differs from SEQ ID NO: 639 by at most one or two amino acids; (viii) an eighth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 640, or an eighth nucleic acid sequence encoding an amino acid sequence that differs from SEQ ID NO: 640 by at most 1, 2, 3, 4, or 5 amino acids; (ix) a ninth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 641, or an amino acid sequence that differs from SEQ ID NO: 641 by at most 1, 2, 3, or 4 amino acids; (x) a tenth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 642, or an amino acid sequence that differs from SEQ ID NO: 642 by at most 1, 2, 3, or 4 amino acids; (xi) an eleventh nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 643, or an eleventh nucleic acid sequence encoding an amino acid sequence differing from SEQ ID NO: 643 by at most one or two amino acids; and (xii) a twelfth nucleic acid sequence encoding the amino acid sequence set forth in SEQ ID NO: 644, or an amino acid sequence that differs from SEQ ID NO: 644 by at most 1, 2, or 3 amino acids; A nucleic acid molecule comprising at least one of the following:

17. 14. The bispecific antibody or antigen-binding fragment thereof of any one of claims 1 to 13 for use in a method for treating a coronavirus infection in a subject.

18. A composition comprising the bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13.

19. 19. The composition of claim 18, further comprising a second medicament for simultaneous, separate, or sequential administration.

20. 20. The composition of claim 19, wherein the second medicament comprises a second antibody or an antigen-binding fragment thereof.

21. 20. The composition of claim 19, wherein the second medicament comprises a bispecific antibody or a bispecific antigen-binding fragment thereof.

22. 20. The composition of claim 18, further comprising a pharmaceutically acceptable excipient or carrier.

23. 19. The composition of claim 18, comprising an effective amount of a bispecific antibody or antigen-binding fragment thereof, a nucleic acid molecule, or a vector, and optionally a pharmaceutically acceptable excipient or carrier, for use in preventing and / or treating a coronavirus infection in a subject, preferably a SARS-CoV-1 or SARS-CoV-2 infection in a subject.

24. 19. The composition of claim 18 for use as a medicine.

25. 19. An inhalation device comprising the composition of claim 18.

26. 14. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, which is bivalent.

27. 14. The bispecific antibody or antigen-binding fragment thereof according to any one of claims 1 to 13, which is capable of binding to at least one stem helix and / or fusion peptide of an α-coronavirus, a β-coronavirus, a γ-coronavirus, and a δ-coronavirus, preferably at least an α-coronavirus and / or a β-coronavirus.

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