Methods for stabilizing henipavirus surface glycoproteins and uses thereof

Abstract

Described are structural identifications of the F and G proteins of a large variety Henipaviruses and their use in creating immunogens useful for vaccine compositions, and in prevention and treatment of Paramyxovirus and Henipavirus infections in a mammal. Also described are methods for screening viral fusion proteins for capability to transition from a pre-fusion to a post-fusion conformation. A monoclonal antibody specific for LayV-F is also provided.

Description

METHODS FOR STABILIZING HENIPAVIRUS SURFACE GLYCOPROTEINS AND USES THEREOF

[0001] This application claims the benefit of and priority to U.S. Provisional Application Nos. 63 / 729,813, filed on December 9, 2024; 63 / 759,426, filed on February 17, 2025; and 63 / 784,714, filed on April 7, 2025, the entire contents each of which are incorporated herein by reference.

[0002] All patents, patent applications and publications cited herein are hereby incorporated by reference in their entirety. The disclosures of these publications in their entireties are hereby incorporated by reference into this application in order to more fully describe the state of the art as known to those skilled therein as of the date of the invention described and claimed herein.

[0003] This patent disclosure contains material that is subject to copyright protection. The copyright owner has no objection to the facsimile reproduction by anyone of the patent document or the patent disclosure as it appears in the U.S. Patent and Trademark Office patent file or records but otherwise reserves any and all copyright rights.SEQUENCE LISTING

[0004] The instant application contains a Sequence Listing which has been submitted electronically in XML format and is hereby incorporated by reference in its entirety. Said XML copy, created on [DATE], is named [NAME] and is [SIZE] bytes in size.BACKGROUND OF THE INVENTION

[0005] Henipaviruses (HNVs) are a genus of ssRNA viruses that includes the highly virulent Nipah virus (NiV) responsible for causing yearly reoccurring outbreaks of deadly disease. The Henipavirus genus is a part of the Paramyxoviridae family, and therefore related to other notable human pathogens such as the parainfluenza viruses, measles, and mumps (1). The combined possibility within the family for both rapid transmission, as with measles, and high lethality, as with NiV, further highlights pandemic risk and necessitates urgent research to establishpreparedness that will allow for a rapid and effective response to a future emergent threat. For HNVs specifically, their identification in diverse animal reservoirs in different geographical locations, their high risk of their zoonotic transmission, and current lack of approved vaccines or therapies to treat HNV infection in humans highlight the high risk and pandemic potential of this genus.

[0006] HNVs have been studied since the discovery of Hendra (HeV) and Nipah viruses in 1994 and 1998, respectively (4-6). Additional members of the genus have been indentified since then, but it was not until 2022 with the discovery of Langya virus (LayV) that another species with pathogenicity in humans was confirmed (2). Like HeV and NiV, LayV was of zoonotic origin. However, unlike HeV, NiV, and most of the other known HNVs at the time, LayV was found to have likely originated from a shrew reservoir, rather than from fruit bats. Several concurrent studies identified more shrew-borne HNVs over a wide geographic range, indicating the existence of a largely uncharacterized clade within the Henipavirus genus (7-9). Among these studies were some untargeted surveillance efforts, revealing new HNV sequences among dozens of other sequences across multiple virus families (10). While the newly discovered shrew-borne species had initially been classified as members of the Henipavirus genus, they have recently been reclassified into the Parahenipavirus genus (3). Herein, we use the abbreviation HNV to refer collectively to species of both genera.

[0007] From the perspective of vaccine or therapeutic countermeasure development for HNV infection, the most important HNV proteins to study are the attachment (G) and fusion (F) surface glycoproteins. These are the two sole surface-exposed HNV proteins, and therefore the only targets for neutralizing antibodies (11). Together, G and F facilitate virus entry into host cells, with G responsible for receptor binding and F mediating membrane fusion. During this process, both the G and F proteins undergo important conformational changes, including the destabilization of the prefusion conformation of F by G. The specifics of these conformational steps, including how G destabilizes the pre-fusion F conformation or “triggers” its pre-fusion to post-fusion transition remain elusive. Although a class I fusion protein (12), the HNV-F protein, along with all Paramyxoviridae F proteins, differs from the canonical class I fusion mechanismin that there is the separation of the receptor attachment mechanism into a separate protein (13). Whereas other viral fusion proteins proceed through the fusion conversion process by repositioning or removal of their attachment subunits, HNV-F proteins retain inherent metastability through F protein architecture alone.

[0008] In addition to mechanistic questions about the HNV surface glycoproteins and their role in virus entry, the antigenicity of these proteins and the possibility of cross-reactivity across the genus remains largely unexplored. While there are several known monoclonal antibodies (mAbs) that are able to neutralize both NiV and HeV (14-18), so far none of these have been found to have any reactivity with LayV (19, 20). Of the few mAbs known to bind to LayV, some have been noted to be cross-reactive with Mojiang virus (Moj V), but not to NiV or HeV (20).SUMMARY OF THE INVENTION

[0009] The continuing emergence of new paramyxovirus species necessitates heightened focus on a family that includes both some of the world's deadliest pathogens in Nipah virus, and most infectious in measles and mumps. Structural determinations of Henipavirus glycoproteins serve as a foundation for future immunogen or therapeutic design.

[0010] The present invention provides an antigenically diverse panel of HNV fusion (F) and attachment (G) glycoproteins from 56 unique HNV strains that reflects global HNV diversity. The F ectodomains and the G head domains were expressed and purified, and their biochemical, biophysical and structural properties were characterized. Immunization experiments in mice were performed leading to the elicitation of antibodies reactive to multiple HNV F proteins.Cryo-EM structures of the diverse F proteins reveal details of pre-fusion state stability and higher order contacts. A crystal structure of the Gamak virus (Gak) G head domain revealed an additional domain added to the conserved 6-bladed, P-propeller fold. Taken together, the present disclosures expand the known structural and antigenic limits of the Henipavirus genus, reveals new cross-reactive epitopes within the HNV genus, and provides foundational data needed for the development of broadly reactive countermeasures.

[0011] By identification of the diversity of Henipaviruses, new strategies are identified that can be utilized for immunogen design. These include identification of a proline residue in the atomic level structure of the Angavokely virus (AngV) fusion (F) protein that stabilizes its prefusion conformation, a variation for the 6-stranded 13-propeller fold identified in a high-resolution crystal structure of the Gamak virus (GakV) G protein head domain that reveals a new minidomain. Furthermore, an epitope is identified that elicits, via vaccination, antibodies that target diverse HNV F proteins. In some embodiments, the proline residue is at amino acid position 447 in AngV.

[0012] In accordance with an embodiment, the present invention provides a method for identification of Henipavirus fusion (F) protein antigens comprising: a) preparing an identification and classification scheme for Henipavirus strains; b) aligning the amino acid sequences of F proteins, wherein the amino acid sequences of the F proteins were truncated at the C-terminal of the ectodomain equivalent to Langya virus residue 447; c) using the alignments of b) to create a phylogenic tree to identify viral strains specific to fruit bat reservoirs and shrew reservoirs; d) preparing a panel of F proteins by preparing DNA expression vectors encoding ectodomain residues spanning residues 1 to the equivalent of 488 in Nipah virus; and e) isolating and purifying the expressed constructs. In some embodiments, step (a) preparing an identificiaton and classification scheme includes one or more of the actions described in Example 1. In some embodiments, preparing an identification scheme can include identifying the sequences to be used. In some embodiments, in step (b), gaps in sequences can be used to optimize sequence alignment.

[0013] In some embodiments, the expression constructs of e) are tested for antigenicity against known antibodies.

[0014] In some embodiments, the expression constructs of e) comprise the amino acid sequences of SEQ ID NOS:70-131 or 132-193.

[0015] In some embodiments, the expression constructs of e) are used to immunize mice to create antibodies.

[0016] In accordance with an embodiment, the present invention provides a method for identification of Henipavirus G protein antigens comprising: a) preparing an identification and classification scheme for Henipavirus strains; b) aligning the amino acid sequences of G proteins, wherein the N-terminal portion of the ectodomains up to the end of the transmembrane domain was deleted, and wherein amino acid sequence of G head domains the amino acid residue analogous to Nipah virus residue 71 was the starting point; c) using the alignments of b) to create a phylogenic tree to identify viral strains specific to fruit bat reservoirs and shrew reservoirs; d) preparing a panel of G proteins by preparing DNA expression vectors encoding the isolated head domain; and e) isolating and purifying the expressed constructs.

[0017] In some embodiments, the expression constructs of e) are tested for antigenicity against known antibodies.

[0018] In some embodiments, the expression constructs of e) are used to immunize mice to create antibodies.

[0019] In accordance with an embodiment, the present invention provides an immunogenic composition comprising a full-length Henipavirus F protein having a proline amino acid inserted at the HRB helix start analogous to the proline in the AngV-F wild type F protein. In some embodiments, the proline residue is at amino acid position 447 in AngV.

[0020] In accordance with an embodiment, the present invention provides an immunogenic composition comprising a Henipavirus F ectodomain protein truncation having a proline amino acid inserted at the HRB helix start analogous to the proline in the AngV-F wild type F protein. In some embodiments, the proline residue is at amino acid position 447 in AngV.

[0021] In accordance with an embodiment, the present invention provides an immunogenic composition comprising a full-length Henipavirus F protein having a proline amino acid inserted at the HRB helix start analogous to the proline in the AngV-F wild type F protein. The sequences shown in Figure 8 are wild-type F protein sequences. In some embodiments, the amino acid equivalent to the amino acid at position 447 in AngV can be substituted for proline. Full-length F proteins having this substitution are SEQ ID NOs. 70-131. Ectodomain-truncated F proteins having this substitution are SEQ ID NOs. 132-193.

[0022] In accordance with an embodiment, the present invention provides an immunogenic composition comprising a Henipavirus F ectodomain protein truncation having a proline amino acid inserted at the HRB helix start analogous to the proline at position 447 in the AngV-F wild type F protein comprising the amino acid sequences as shown in SEQ ID NOS: 132-193

[0023] In accordance with an embodiment, the present invention provides an antibody which is capable of binding the pre-fusion LayV-F protein identified as 22F5 comprising a heavy chain variable region (SEQ ID NO: 68) and a light chain variable region (SEQ ID NO: 69).

[0024] Other embodiments are disclosed herein.BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.Figure 1: Flowchart and Tree. Shows a flowchart that describes the origin of sequences used and the filtering mechanisms, genome and F / G sequence completion, used to assemble a list of nonredundant F and G proteins. Tree: A phylogenetic tree based on full-genome nucleotide sequences of the selected strains. Horizontal branch lengths are to scale to represent sequence divergence. Hollow boxes are drawn to represent broad groupings within the tree such as genus, with Classi cal / Bat Henipaviruses (green) and Shrew / Parahenipaviruses (orange), as well as potential species groupings such as Nipah viruses, (dark red) and Hendra viruses (dark blue). Solid highlight boxes are drawn to represent more narrow groupings of specific species or subspecies groupings such as Nipah-Malaysia (yellow), Nipah-Bangladesh (purple), and Langya- China (tan). The tips of each branch are labelled with the strain names and associated gen bank accession number. Each tip is colored based on which unique glycoproteins are available from that strain: unique F ectodomain only, blue, unique G ectodomain only, yellow, no unique G head domain, dashed redborder, both unique F and G ectodomain, green.Figures 2A-2D: Purification and Antigenic Characterization of Henipavirus Fusion Proteins. (2A) Representative SEC profiles of HNV-F Proteins with peaks annotated. Theoligomer-of-trimer peak, sometimes called the “middle peak,” and the main trimer peaks are collected separately. NiVop08 is a pre-fusion stabilized construct 31 not part of the panel but purified for a vaccination study and for comparison to wild-type proteins. An inset to the right shows mass photometry analysis of NiV-Ml-F SEC components. Each individual measurement is assigned a calculated molecular weight by the instrument software, and individual results are binned in 10 kDa bin sizes. The graph reports the count of events for each molecular weight bin with the main, middle, and mixed peak data overlayed. (2B) Graphical timeline of a vaccination study in mice using HNV-F antigens. Each group was vaccinated with LayV-Cl-F and boosted with LayV-Cl-F or NiV antigens as marked. Blood draws and antigen injections occurred at the indicated weeks after the start of the study. The weeks with an asterisk, with the syringe and blood drop marked with F are weeks where a fusion was performed on an individual mouse. (2C) Heatmap of antibody binding to HNV-F proteins, on a spectrum of blue for low binding to red for high binding. The left-side graph is based on ELISA data with immobilized F proteins, antibody analytes, and anti-Human Fabconjugated HRP as a reporter. The values for each combination are based on the log of the area under the curve for the plotted absorbance over increasing concentrations. The right-side graph is based on Bio-layer interferometry (BLT) with immobilized F proteins and direct measurement of analyte to the BLI sensors, either mouse Fabs or mouse IgG (22F5 and 1C8). The values for each combination are based on the maximum wavelength shift measured during the association phase of the BLI experiment. HNV-F antigens colored as in Figure 1, antibody analytes colored as follows: green, known anti-NiV-F binding; salmon, known anti-Lay V-F binding; gray, anti-influenza HA negative control antibody (ELISA only). (2D) Surface plasmonic resonance (SPR) analysis of 22F5 and 4G5 antibody competition for binding of a series of Lay V-F constructs in a variety of conformational states. Solid lines represent 22F5 alone and dashed lines with 4G5.

[0026] Figures 3A-3E: Structure and Binding of 22F5 Fab with Lay V-F Ectodomain. (3 - 3B). Cryo-EM maps of 22F5 Fab bound to post-fusion LayV-l-F (3A) or pre-fusion LayV-F-DS (3B) with domains colored and labelled. (3C) Zoomed-in views of the models (Left: PostFusion, Right: Pre-Fusion) built from the cryo-EM maps colored by domain or antibody CDRs.(3D) Polar contacts between 22F5 Fab and the post-fusion LayV-l-F. Interacting residues labeled and shown in stick representation, with the color scheme continued from 3C. (3E) SPR measurement of affinity and kinetics of binding of 22F5 Fab to LayV-F ectodomains. The black lines in the graphs are the blank subtracted sensorgrams and the red lines are the fit of the data to a 1 : 1 Langmuir binding model. The table to the right shows the affinity and kinetic parameters obtained from the measurements.

[0027] Figures 4A-4F: Thermostability of Henipavirus F Proteins. (4A) DSF profiles of a set of HNV proteins showing the first derivative of 350 / 330 nm emission ratio as a function of temperature. Dotted lines of the same color represent the DSF profile of the same proteins after first being incubated at 60°C for 90 minutes. (4B) Two-dimensional NSEM class averages of a portion of the heat-treated samples from A. (4C) DSF profiles of diverse HNV-F species. Sections of the profile are noted indicating the conformational steps occurring at those temperatures, with peaks of interest highlighted. (4D) A subset of DSF profiles from 4C, containing only those species that do not demonstrate any noticeable conformational conversion peak. Below are representative two-dimensional NSEM class averages for select members of this subset. (4E) A subset of DSF profiles from 4C, containing only those species that demonstrate some presence of the standard conformational conversion signal. Below are representative two- dimensional NSEM class averages for select members of this subset. (4F) A subset of DSF profiles from 4C, containing only those species that demonstrate a profile notably different from the typical HNV-F species. Below are representative two-dimensional NSEM class averages for the two members of this subset.

[0028] Figures 5A-5E: Cryo-EM structures of diverse Henipavirus F proteins. (5A) Cryo- EM structure of the AngV-F ectodomain. Three different particle populations in the AngV-F and their representative 2D classes. Cryo-EM reconstruction of top and side view shown for AngV F trimer (left), dimer-of-trimers (middle) and hexameric lattice (right). (5B) Comparison of the AngV-F, Lay V and NiV F proteins. Top. A slice through the F-protein visualizing the central cavity and the placement of the stalk region within the F-protein. Bottom. A top-down view showing measurements. (5C) Zoom-in view of the stalk region featuring amino acid present atthe start of helix bundle (top) and their varied electrostatic potentials (bottom). (5D) AngV-F cryo-EM structure colored by protomer with the glycans shown as spheres and colored by element. Glycans at the different regions (Apex, non-canonical NNV, stalk) are marked with arrows. (5E) The dimer-of-trimer interaction interface is shown (top), along with their atomic- level interactions (bottom).

[0029] Figures 6A-6D: Translatable Pre-fusion Stabilization of Henipavirus FProteins. (6A) Location of HRB proline mutation in select HNV-F proteins. Left: Site of NiV K453P mutation shown as red spheres in the HNV-F ectodomain structure, with protomers colored in shades of blue. Right: A sequence alignment of the residues surrounding the HRB proline mutation, colored by residue type: hydrophobic, orange, polar, pink, negative charge, red, aromatic, white, conformationally significant, green, HRB proline mutation, dark green. (6B-6D) Characterization of the HRB proline mutation in three HNVF species, (6B) NiV-Ml-F, (6C) SHNV5-1-F, (6D) GakV-l-F. Top: SEC overlay of the WT (right axis) and mutant (left axis) with mAU values scaled to a IL transfection volume. Middle: DSF analysis overlay of the WT and mutant. The middle line represents the average with standard deviation represented with thin lines above and below the average line and shaded in-between. Bottom: Comparison of representative NSEM 2D classes from WT (top) and mutant (Bottom).

[0030] Figures 7A-7G: Structure and Characterization of Henipavirus G Proteins. (7A) Heatmap of recombinant Fc-Ephrin-B2 and B3 binding to G head domains, on a spectrum of blue for low binding to red for high binding. The graph is based on BLI data with immobilized Ephrin and G head domain analytes. The value for each combination is based on the maximum response value from the association step. G head domain antigens in the table colored by group as in Figure 1. (7B) Heatmap of antibody binding to G head domains, with color scheme identical to (7 A). The graph is based on BLI data as described in (7A) using immobilized anti-G antibodies. (7C) Sequence alignment of the C terminal end of select HNV-G proteins. (7D) Three views of the GakV-G head domain colored in rainbow from N to C terminal. (7E) Superposition of GakV-G head domain on Ephrin-bound NiV-G head domain (PDB: 2VSM 51). GakV-G is colored gray, NiV-G cyan and Ephrin light blue. Ephrin is shown as a transparentsurface. The GakV-G glycan 506 is shown as blue spheres. (7F) Topology diagram of GakV-G protein with blades of the beta propeller colored in rainbow from blade 1 through 6. (7G) Left: Two views of the GakV-G head domain colored in rainbow with the N terminal Asp to His218 shown as a transparent surface. The additional minidomain defined in the GakV-2-G head structure is highlighted within a dashed oval. Right: G head domains of Lay V and NiV.

[0031] Figures 8A-8E: Sequence Alignments of HNV-F Proteins. The sequences are wildtype (no proline substitution at the position equivalent to position 447 in AngV), full-length F proteins. In order, these sequences are SEQ ID NOs: 1-35.

[0032] Figures 9A-9F: Sequence Alignments of HNV-G Proteins. The sequences are wild-type G proteins. In order, these are SEQ ID NOs: 36-67.

[0033] Figures 10A-C: A table of the viruses tested and originsDETAILED DESCRIPTION OF THE INVENTION

[0034] Whether there are any broadly reactive anti-HNV antibodies or where the limits of cross-reactivity are within the genus considering newly-discovered species remains unknown. To address these genus-wide mechanistic and antigenic questions, we assembled a curated set of HNV G and F sequences that samples the diversity of the genus as broadly as possible in order to purify and characterize these proteins, biochemically, biophysically, and structurally.

[0035] Disclosed herein is a sequence and biophysical analysis of F and G proteins from Henipaviruses. In some embodiments, these studies have led to determination that a proline substitution can stabilize F proteins from these viruses in a pre-fusion conformation. In some embodiments, these studies have led to determination of epitopes that elicits antibodies that target diverse F proteins.

[0036] Over the past few years, there has been a rapid expansion of identified species and strains in the Henipavirus genus. Given the history of zoonotic transmission with these viruses, characterization of these new species is a first step for pandemic readiness. Up until recently, the strains in our expanded panel of Henipaviruses were uncharacterized and their relationships to other Henipaviruses unknown. Now, we have been able to identify two new species groupingscontaining strains that were identified independently. Given that the two strains of Daeryong virus were from sample collection in two different countries (Korea and China), this indicates that, just as with Nipah, there is a likely a broad geographic range of these newly-identified, shrew-borne Henipaviruses.

[0037] Purifying and characterizing the F protein ectodomains of these strains has revealed that despite low sequence identity, HNV-F proteins share many architectural and biophysical similarities, a trend we previously noted by comparing Langya virus to Niaph virus (18), and one that is apparent with our species panel This was apparent from the SEC profiles. The formation of these complexes is transient and dependant on the concentration and environment of the F protein, as indicated by the almost exclusive presence of trimer molecular weights detected by mass photometry. The ubiquity of this type of interaction indicates that oligomerization of F proteins likely is a conserved trait within Henipaviruses.

[0038] In the case of Angavokely virus, the formation of a hexamer lattice of F protein trimers, was observed in the cryo-EM datasets. The existence of a hexamer-of-trimers arrangment has been noted previously with NiV (37). Our structure demonstrates the ability of AngV-F to multimerize in this way without any such assistance, and, critically, without the involvemnt of the transmembrane domain, viral envelope, cytoplasmic domain, or viral matrix protein. The hexameric lattice is formed through ectodomain interactions.

[0039] Previous studies have defined the lattice of Paramyxoviridae and Pneumoviridae F proteins, with various arrangements being determined. In one case, hexameric lattices have been observed, like with parainfluenza virus (36). In others, such as with measles virus and respiratory syncytial virus, alternate arrangements of fusion and attachment proteins have been observed (Refs). The results with AngV-F indicate that HNV-F proteins may adopt the hexameric arrangement. Furthermore, while not wishing to be held to any theory, it may be that the formation of a hexameric F lattice may comprise one stage of HNV-F functionality.

[0040] The antigenicity of new HNV strains had not yet been explored. By assessing the binding of a panel of both previously identified anti-HNV antibodies, other broadly-reactive antibodies, and newly discovered antibodies elicited through vaccination, we can now betterdescribe the antigenic landscape of HNV proteins. We saw little ability of anti-NiV / HeV antibodies to bind to non-NiV / HeV proteins, with the exception of 1E5. We found antigenic overlap between the F proteins of LayV, MojV, SHNV5, and to a lesser extent, GakV. Both a previously reported anti-MojV-F antibody, 4G5, and our vaccine-elicited antibodies, 22F5 especially, bound to these strains. When considering our phylogenetic analysis, both by full genome and amino acid sequence, it can be seen this cross-reactivity spans distantly related strains. SHNV5 is more distantly related from LayV than HeV is from NiV. GakV is more distant phylogenetically, appearing to be roughly as distant from LayV as CedV is from NiV.

[0041] While not wishing to be held to any theory, with a group of viruses prone to such sequence diversity, one major concern in the event of a possible widespread outbreak of disease in humans is that mutations would occur so quickly that the immunity conferred from prior infection or immunization would not be able to track with newer strains. This phenomenon can be observed with yearly outbreaks of new influenza strains or with the progression of the SARS- CoV-2 pandemic, where new strains emerged and quickly dominated the pool of circulating virus as they evaded prior immunity. A focus of efforts to address this possibility has been designing immunogens that can elicit antibodies with broad coverage of the currently existing and future diversity of strains. Here we see that there is demonstrated cross-reactivity among a large portion of the shrew-borne clade (Parahenipavirus).

[0042] Herein biophysical characterizations of HNV-F proteins has provided a method by which we can assess their metastability. The pre- and postfusion conformations of HNV-F proteins are very visually distinct, allowing for relatively simple assignment of conformational states in a structural dataset. However, the process of obtaining structural data for such a large panel can be expensive and time-consuming. Therefore, we worked to develop our DSF-based technique to indicate the conformational state without relying on structure determination. In an intrinsic protein fluorescence DSF experiment, emission wavelength of key residues change based on their chemical environment, which changes during refolding or unfolding processes. For F proteins, both the conformational steps associated with pre- to postfusion conformational conversion and denaturation and dissociation of the trimer would be likely to be detected byDSF. However, without any additonal information, changes in DSF signal could not clearly be assigned to a particular conformational event. By combining the concept of heat treatment, previously used to force conversion with PIV3 (36), with DSF, we force conversion on command. By seeing elimination of early negative peaks only for wild-type proteins, we confirmed that this peak corresponds to conformational conversion. By applying this technique to the entire panel, we linked differences in sequence or structure to more definitive conclusions about metastability.

[0043] In the case of AngV-F, the differences between its structure and those of other HNV- F proteins line up with the differences observed in its DSF profile. AngV-F has greater metastability than other HNV-F proteins. Introduction of a Proline residue at the beginning of the stalk domain, also referred to as the HRB region, is responsible for this change. While not wishing to be held to any theory, during conformational conversion in most HNV-F proteins, it is likely that this charge cluster works to disperse the helix bundle, serving to lower the energetic barrier to conformational conversion. Instead, in AngV-F, no such electrostatic repulsion is likely at this site, leading to a more stable prefusion HRB arrangement, which is likely why the AngV-F stalk domain is so well resolved in our Cryo-EM structure.

[0044] DSF analysis of G head domains revealed a diversity of themostability patterns. Unlike F proteins, the head domains are a simple and stable fold that is not known to have any major internal conformational rearrangements as part of its receptor-binding mechanism. Some studies have indicated that receptor binding by HNV-G proteins could effect subtle changes in protein flexibility that don’t manifest as large-scale conformational events (47). From our crystal structure of the Gamak virus head domain, we observed likely sources of mechanistic differences between species. Specifically, the presence of a glycan in the face of the beta propellor analagous to the binding site for NiV-G indicates that it is unlikely a protein binder similar to EFNB2 / B3 which binds at this side in GakV.

[0045] Ultimately, this broad characterization of HNV proteins provides a set of valuable biophysical and biochemical data for many species and strains that, to this point, had only been known as a sequence in a database.

[0046] Unless defined otherwise, all technical and scientific terms used herein have the meaning commonly understood by a person skilled in the art to which this invention belongs. The following references provide one of skill with a general definition of many of the terms used in this invention: Singleton et al., Dictionary of Microbiology and Molecular Biology (2nd ed. 1994); The Cambridge Dictionary of Science and Technology (Walker ed., 1988); The Glossary of Genetics, 5th Ed., R. Rieger et al. (eds.), Springer Verlag (1991); and Hale & Marham, The Harper Collins Dictionary of Biology (1991). As used herein, the following terms have the meanings ascribed to them below, unless specified otherwise.Definitions

[0047] Ranges provided herein are understood to be shorthand for all of the values within the range. For example, a range of 1 to 50 is understood to include any number, combination of numbers, or sub-range from the group consisting 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50.

[0048] As used herein, the terms “treat,” treating,” “treatment,” and the like can refer to reducing or ameliorating a disorder and / or symptoms associated therewith. It will be appreciated that, although not precluded, treating a disorder or condition does not require that the disorder, condition or symptoms associated therewith be completely eliminated.

[0049] Unless specifically stated or obvious from context, as used herein, the term "or" is understood to be inclusive. Unless specifically stated or obvious from context, as used herein, the terms "a", "an", and "the" are understood to be singular or plural.

[0050] Unless specifically stated or obvious from context, as used herein, the term “about” is understood as within a range of normal tolerance in the art, for example within 2 standard deviations of the mean. About can be understood as within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value. Unless otherwise clear from context, all numerical values provided herein are modified by the term about.

[0051] Any compositions or methods provided herein can be combined with one or more of any of the other compositions and methods provided herein.

[0052] As used herein, the terms “prevent,” “preventing,” “prevention,” “prophylactic treatment” and the like can refer to reducing the probability of developing a disorder or condition in a subject, who does not have, but is at risk of or susceptible to developing a disorder or condition.

[0053] Wherever any of the phrases “for example,” “such as,” “including” and the like are used herein, the phrase “and without limitation” is understood to follow unless explicitly stated otherwise. Similarly, “an example,” “exemplary” and the like are understood to be nonlimiting.

[0054] The term “substantially” allows for deviations from the descriptor that do not negatively impact the intended purpose. Descriptive terms are understood to be modified by the term “substantially” even if the word “substantially” is not explicitly recited.

[0055] The terms “comprising” and “including” and “having” and “involving” (and similarly “comprises”, “includes,” “has,” and “involves”) and the like are used interchangeably and have the same meaning. Specifically, each of the terms is defined consistent with the common United States patent law definition of “comprising” and is therefore interpreted to be an open term meaning “at least the following,” and is also interpreted not to exclude additional features, limitations, aspects, etc. Thus, for example, “a process involving steps a, b, and c” means that the process includes at least steps a, b and c. Wherever the terms “a” or “an” are used, “one or more” is understood, unless such interpretation is nonsensical in context.

[0056] As used herein, the term “about” can refer to approximately, roughly, around, or in the region of. When the term “about” is used in conjunction with a numerical range, it modifies that range by extending the boundaries above and below the numerical values set forth. The term “about” is used herein to modify a numerical value above and below the stated value by a variance of 20 percent up or down (higher or lower).

[0057] The term "administering" can refer to introducing a substance into a subject. Any route of administration can be utilized including, for example, intranasal, topical, oral, parenteral, intravitreal, intraocular, ocular, subretinal, intrathecal, intravenous, subcutaneous,transcutaneous, intracutaneous, intracranial and the like administration. For example, “parenteral administration” can refer to administration via injection or infusion. Parenteral administration includes, but is not limited to, subcutaneous administration, intravenous administration, and intramuscular administration. For example, the inhibitor and / or degrader can be administered intranasally, by inhalation, intrapulmonarily, or by injection (e.g., intravenous or subcutaneous). Herein, administering can refer to introducing an epitope scaffold or composition thereof into a subject. In some embodiments, the purpose of the administration is prophylactic protection against coronavirus infection and / or symptoms of disease caused by coronavirus infection.

[0058] The term “simultaneous administration” can refer to a first agent and a second agent administered less than about 15 minutes apart, e.g., less than about 10, 5, or 1 minutes. When the first agent and the second agent are administered simultaneously, the first and second treatments can be in the same composition (e.g., a composition comprising both the first and second therapeutic agents) or separately (e.g., the first therapeutic agent is contained in one composition and the second treatment is contained in another composition).

[0059] The term “sequential administration” can refer to a first agent and a second agent administered to a subject greater than about 15 minutes apart, such as greater than about 20, 30, 40, 50, 60 minutes, or greater than 60 minutes apart. Either agent can be administered first. For example, the first agent and the second agent can be included in separate compositions, which can be included in the same or different packages or kits.

[0060] The terms “co-administration” or the like, as used herein, can refer to the administration of a first active agent and at least one additional active agent to a single subject, and is intended to include treatment regimens in which the compounds and / or agents are administered by the same or different route of administration, in the same or a different dosage form, and at the same or different time.

[0061] As used herein, the term “domain” can refer to a functional portion, segment or region of a protein or polypeptide. “Interaction domain” can refer to a portion, segment or region of a protein, polypeptide or protein fragment that is responsible for the physical affinity of thatprotein, protein fragment or isolated domain for another protein, protein fragment or isolated domain.

[0062] The term “in combination” can refer to the use of more than one therapies (e.g., one or more prophylactic and / or therapeutic agents). The use of the term “in combination” does not restrict the order in which therapies are administered to a subject with a disease or disorder, or the route of administration.

[0063] The term “epitope” can refer to a protein determinant capable of specific binding to an immunoglobulin, a scFv, a T-cell receptor and the like.

[0064] The term “grafting” can refer to combining an epitope with a protein scaffold. Generally, as described herein, computational methods can be used to identify protein scaffolds that have regions that have the same shape or conformation as a selected epitope (e.g., the scaffolds have regions that can display the epitope such that an immune response to a desired conformation of the epitope can be produced). “Grafting” of the epitope into the protein scaffold at this region (e.g., the epitope can replace the same shape / conformation segment of the protein scaffold) produces the epitope scaffold. “Transplanting” can used instead of grafting.

[0065] As used herein, the term “immunogen” and related terms “immunogenic” refer to molecules that have the ability to induce an immune response, including antibodies and / or cellular immune responses in an animal, e.g., a mammal. Although an immunogen may be antigenic, an “antigen” need not necessarily be an “immunogen” because such molecules may not induce a sufficient immune response. In some examples, this may be because of the antigen’s size, conformation and the like. In some embodiments, an immunogenic composition can contain one or more immunogens that can induce an immune response that can specifically recognize viruses that contain the immunogen or antigen.

[0066] As used herein, the terms "immunological binding," and "immunological binding properties" can refer to non-covalent interactions of the type which occur between an immunoglobulin molecule and an antigen for which the immunoglobulin is specific. The strength, or affinity of immunological binding interactions can be expressed in terms of the equilibrium binding constant (Ka) of the interaction, wherein a smaller Ka represents a greateraffinity. Immunological binding properties of selected polypeptides can be quantified using methods well known in the art. One such method entails measuring the rates of antigen- binding site / antigen complex formation and dissociation, wherein those rates depend on the concentrations of the complex partners, the affinity of the interaction, and geometric parameters that equally influence the rate in both directions. Thus, both the "on rate constant" (Kon) and the "off rate constant" (Koir) can be determined by calculation of the concentrations and the actual rates of association and dissociation. The ratio of Koir / Konallows the cancellation of all parameters not related to affinity, and is equal to the equilibrium binding constant, KD.

[0067] The terms “prevent,” “preventing” and / or “prevention” can refer to the prevention of the onset, recurrence or spread of a disease or disorder, or of one or more symptoms thereof. In certain embodiments, the terms can refer to the treatment with or administration of a compound provided herein, with or without other additional active compound, prior to the onset of symptoms, particularly to patients at risk of diseases or disorders provided herein. The terms encompass the inhibition or reduction of a symptom of the particular disease. For example, one or more of the following effects can result from the administration of a therapy or a combination of therapies as described herein: (i) the inhibition of the development or onset of a viral infection and / or a symptom associated therewith; and (ii) the inhibition of the recurrence of a viral infection and / or a symptom associated therewith.

[0068] The term “zw vivo" can refer to an event that takes place in a subject's body.

[0069] The term “zzz vitro" can refer to an event that takes places outside of a subject's body.

[0070] The term “ex vivo" can refer to outside a living subject. Examples of ex vivo cell populations include in vitro cell cultures and biological samples such as fluid or tissue samples from humans or animals. Such samples can be obtained by methods well known in the art. Exemplary biological fluid samples include blood, cerebrospinal fluid, urine, saliva. Exemplary tissue samples include tumors and biopsies thereof. In this context, the present compounds can be in numerous applications, both therapeutic and experimental.

[0071] The terms “manage,” “managing,” and “management,” in the context of the administration of a therapy to a subject, can refer to the beneficial effects that a subject derivesfrom a therapy, which does not result in a cure of a viral infection. In embodiments, a subject is administered one or more therapies to manage a viral infection so as to prevent the progression or worsening of the viral infection.

[0072] The phrase "pharmaceutical composition" or a “pharmaceutical formulation” can refer to a composition or pharmaceutical composition suitable for administration to a subject, such as a mammal, especially a human and that can refer to the combination of an active agent(s), or ingredient with a pharmaceutically acceptable carrier or excipient, making the composition suitable for diagnostic, therapeutic, or preventive use in vitro, in vivo, or ex vivo. A “pharmaceutical composition” can be sterile and can be free of contaminants that can elicit an undesirable response within the subject (e.g., the compound(s) in the pharmaceutical composition is pharmaceutical grade). Pharmaceutical compositions can be designed for administration to subjects or patients in need thereof via a number of different routes of administration including oral, intranasal, topical, intravenous, buccal, rectal, parenteral, intraperitoneal, intradermal, intratracheal, intramuscular, subcutaneous, by stent-eluting devices, catheters-eluting devices, intravascular balloons, inhalational and the like. In some embodiments, an immunogenic composition is a type of pharmaceutical composition.

[0073] As used throughout the present application, the term "polypeptide” is used in its broadest sense to refer to a sequence of subunit amino acids. The polypeptides described herein may be chemically synthesized or recombinantly expressed. Polypeptide can encompass a singular “polypeptide” as well as plural “polypeptides,” and can refer to a molecule composed of monomers (amino acids) linearly linked by amide bonds (also known as peptide bonds). The term “polypeptide” can refer to any chain or chains of two or more amino acids and does not refer to a specific length of the product. Thus, peptides, dipeptides, tripeptides, oligopeptides, “protein,” “amino acid chain,” or any other term used to refer to a chain or chains of two or more amino acids, can refer to “polypeptide” herein, and the term “polypeptide” can be used instead of, or interchangeably with any of these terms. “Polypeptide” can also refer to the products of post-expression modifications of the polypeptide, including without limitation glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting / blocking groups,proteolytic cleavage, or modification by non-naturally occurring amino acids. A polypeptide can be derived from a natural biological source or produced by recombinant technology but is not necessarily translated from a designated nucleic acid sequence. It can be generated in any manner, including by chemical synthesis.

[0074] Peptides, polypeptides and proteins can be said to have amino acid “substitutions.” Such substitutions can refer to replacement of an amino acid at a specific position in a peptide, polypeptide or protein with a different amino acid. In some embodiments, an amino acid substation is said to be “conservative.” A “conservative amino acid substitution” can be a substitution in which an amino acid residue is replaced with an amino acid residue having a similar side chain. Families of amino acid residues having similar side chains have been defined in the art, including basic side chains (e.g., lysine, arginine, histidine), acidic side chains (e.g., aspartic acid, glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, methionine, tryptophan), beta-branched side chains (e.g., threonine, valine, isoleucine) and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, histidine). In some embodiments, families of amino acid side chains can be grouped, as known in the art, using other properties or characteristics.

[0075] “Specifically binds” or “has specificity to,” can refer to an antibody that binds to an epitope via its antigen-binding domain, and that the binding entails some complementarity between the antigen-binding domain and the epitope. For example, an antibody is said to “specifically bind” to an epitope when it binds to that epitope, via its antigen-binding domain more readily than it would bind to a random, unrelated epitope.

[0076] The term “subject,” “patient” or “individual” can refer to any organism to which aspects of the invention can be administered, e.g., for experimental, diagnostic, prophylactic, and / or therapeutic purposes. For example, subjects to which compounds of the disclosure can be administered include animals, such as mammals. Non-limiting examples of mammals include primates, such as humans. For veterinary applications, a wide variety of subjects will be suitable, e.g., livestock such as cattle, sheep, goats, cows, swine, and the like; poultry such as chickens,ducks, geese, turkeys, and the like; and domesticated animals for example pets such as dogs and cats. For diagnostic or research applications, a wide variety of mammals will be suitable subjects, including rodents (e.g., mice, rats, hamsters), rabbits, primates, and swine such as inbred pigs and the like. The term “living subject” can refer to a subject noted above or another organism that is alive. The term “living subject” can refer to the entire subject or organism and not just a part excised (e.g., a liver or other organ) from the living subject.

[0077] The terms “therapies” and / or “therapy” can refer to any protocol(s), method(s), compositions, formulations, and / or agent(s) that can be used in the prevention, treatment, management, or amelioration of a viral infection or a symptom associated therewith. In embodiments, the terms “therapies” and “therapy” can refer to biological therapy, supportive therapy, and / or other therapies useful in treatment, management, prevention, or amelioration of a viral infection or a symptom associated therewith known to one of skill in the art.

[0078] The terms “therapeutic agent”, and “therapeutic agents” can refer to any agent(s) which can be used in the prevention, treatment and / or management of a viral infection or a symptom associated therewith.

[0079] The term "therapeutically effective amount" can refer to that amount of an embodiment of the composition or pharmaceutical composition being administered that will relieve to some extent one or more of the symptoms of the disease or condition being treated, and / or that amount that will prevent, to some extent, one or more of the symptoms of the condition or disease that the subject being treated has or is at risk of developing.

[0080] The terms “treat,” “treatment,” and “treating” can refer to the management and care of a subject for the purpose of combating a condition, disease or disorder, such as a viral infection, in any manner in which one or more of the symptoms of a disease or disorder are ameliorated or otherwise beneficially altered. The term can include the full spectrum of treatments for a given condition from which the patient is suffering, such as administration of the active compound for the purpose of: alleviating or relieving symptoms or complications; delaying the progression of the condition, disease or disorder; curing or eliminating the condition, disease or disorder; and / or preventing the condition, disease or disorder, wherein"preventing" or "prevention" can refer to the management and care of a patient for the purpose of hindering the development of the condition, disease or disorder, and includes the administration of the active compounds to prevent or reduce the risk of the onset of symptoms or complications.

[0081] A “variant” can refer to a virus having one or more mutations as compared to a known virus. A strain can be a genetic variant or subtype of a virus. The terms “strain”, “variant,”, and “isolate” may be used interchangeably. In certain embodiments, a variant has developed a “specific group of mutations” that causes the variant to behave differently than that of the strain it originated from.

[0082] The term “viral infection” can refer to the invasion by, multiplication and / or presence of a virus in a cell or a subject.

[0083] In one embodiment, a viral infection can be an “active” infection. An active infection can refer to one in which the virus is replicating in a cell or a subject. Active infections can be characterized by the spread of the virus to other cells, tissues, and / or organs, from the cells, tissues, and / or organs initially infected by the virus.

[0084] In embodiments, the viral infection can be a “latent” infection. A latent infection can refer to one in which the virus is not replicating. In some embodiments, an infection can refer to the pathological state resulting from the presence of the virus in a cell or a subject, or by the invasion of a cell or subject by the virus.

[0085] The term “protein scaffold” or “scaffold polypeptide” can refer to a molecule that can be a “framework” for or that can “host” an epitope. A protein scaffold containing a “grafted epitope” is called an epitope scaffold or ES.

[0086] The term “grafting” can refer to combining an epitope with a protein scaffold. Generally, as described herein, computational methods can be used to identify protein scaffolds that have regions that have the same shape or conformation as a selected epitope (e.g., the scaffolds have regions that can display the epitope such that an immune response to the epitope can be produced). “Grafting” of the epitope into the protein scaffold at this region (e.g., the epitope can replace the same shape / conformation segment of the protein scaffold) produces the epitope scaffold. “Transplanting” can used instead of grafting.Antibodies

[0087] An “antibody” or “antigen-binding polypeptide” can refer to a polypeptide or a polypeptide complex that specifically recognizes and binds to an antigen, such as fusion peptide. An antibody can be a whole antibody and any antigen binding fragment or a single chain thereof. For example, “antibody” can include any protein or peptide containing molecule that comprises at least a portion of an immunoglobulin molecule having biological activity of binding to the antigen. Non-limiting examples a complementarity determining region (CDR) of a heavy or light chain or a ligand binding portion thereof, a heavy chain or light chain variable region, a heavy chain or light chain constant region, a framework (FR) region, or any portion thereof, or at least one portion of a binding protein. As used herein, the term "antibody" can refer to an immunoglobulin molecule and immunologically active portions of an immunoglobulin (Ig) molecule, i.e., a molecule that contains an antigen binding site that specifically binds (immunoreacts with) an antigen. By "specifically binds" or "immunoreacts with" is meant that the antibody reacts with one or more antigenic determinants of the desired antigen and does not react with other polypeptides.

[0088] The terms “antibody fragment” or “antigen-binding fragment” can refer to a portion of an antibody such as F(ab')2, F(ab)2, Fab', Fab, Fv, scFv and the like. Regardless of structure, an antibody fragment binds with the same antigen that is recognized by the intact antibody. The term “antibody fragment” can include aptamers (such as spiegelmers), minibodies, and diabodies. The term “antibody fragment” can also include any synthetic or genetically engineered protein that acts like an antibody by binding to a specific antigen to form a complex. Antibodies, antigen-binding polypeptides, variants, or derivatives described herein include, but are not limited to, polyclonal, monoclonal, multispecific, human, humanized or chimeric antibodies, single chain antibodies, epitope-binding fragments, e.g., Fab, Fab' and F(ab')2, Fd, Fvs, single-chain Fvs (scFv), single-chain antibodies, dAb (domain antibody), minibodies, disulfide-linked Fvs (sdFv), fragments comprising either a VL or VH domain, fragments produced by a Fab expression library, and anti-idiotypic (anti-Id) antibodies.

[0089] A “single-chain variable fragment” or “scFv” can refer to a fusion protein of the variable regions of the heavy (VH) and light chains (VL) of immunoglobulins. A single chain Fv ("scFv") polypeptide molecule is a covalently linked VH:VL heterodimer, which can be expressed from a gene fusion including VH- and VL-encoding genes linked by a peptide- encoding linker. (See Huston et al. (1988) Proc Nat Acad Sci USA 85(16): 5879-5883). In embodiments the regions are connected with a short linker peptide, such as a short linker peptide of about ten to about 25 amino acids. The linker can be rich in glycine for flexibility, as well as serine or threonine for solubility, and can either connect the N-terminus of the VH with the C- terminus of the VL, or vice versa. This protein retains the specificity of the original immunoglobulin, despite removal of the constant regions and the introduction of the linker. A number of methods have been described to discern chemical structures for converting the naturally aggregated, but chemically separated, light and heavy polypeptide chains from an antibody V region into an scFv molecule, which will fold into a three-dimensional structure substantially similar to the structure of an antigen-binding site. See, e.g., U.S. Patent No. 5,091,5 13; No. 5,892,019; No. 5,132,405; and No. 4,946,778, each of which are incorporated by reference in their entireties.

[0090] Very large naive human scFv libraries have been and can be created to offer a large source of rearranged antibody genes against a plethora of target molecules. Smaller libraries can be constructed from individuals with infectious diseases in order to isolate disease-specific antibodies. (See Barbas et al., Proc. Natl. Acad. Sci. USA 89:9339-43 (1992); Zebedee et al, Proc. Natl. Acad. Sci. USA 89:3 175-79 (1992)).

[0091] Antibody molecules obtained from humans fall into five classes of immunoglubulins: IgG, IgM, IgA, IgE and IgD, which differ from one another by the nature of the heavy chain present in the molecule. Those skilled in the art will appreciate that heavy chains are classified as gamma, mu, alpha, delta, or epsilon (y, p, a, 8, s) with some subclasses among them (e.g., yl-y4). Certain classes have subclasses as well, such as IgGl, IgG2, IgG3 and IgG4 and others. The immunoglobulin subclasses (isotypes) e.g., IgGl, IgG2, IgG3, IgG4, IgG5, etc. are well characterized and are known to confer functional specialization. With regard to IgG, a standardimmunoglobulin molecule comprises two identical light chain polypeptides of molecular weight approximately 23,000 Daltons, and two identical heavy chain polypeptides of molecular weight 53,000-70,000. The four chains are typically joined by disulfide bonds in a “Y” configuration wherein the light chains bracket the heavy chains starting at the mouth of the “Y” and continuing through the variable region. Immunoglobulin or antibody molecules described herein can be of any type (e.g., IgG, IgE, IgM, IgD, IgA, and IgY), class (e.g., IgGl, IgG2, IgG3, IgG4, IgAl and IgA2) or subclass of an immunoglobulin molecule.

[0092] Light chains are classified as either kappa or lambda (K, I). Each heavy chain class can be bound with either a kappa or lambda light chain. In general, the light and heavy chains are covalently bonded to each other, and the “tail” portions of the two heavy chains are bonded to each other by covalent disulfide linkages or non-covalent linkages when the immunoglobulins are generated either by hybridomas, B cells, or genetically engineered host cells. In the heavy chain, the amino acid sequences run from an N-terminus at the forked ends of the Y configuration to the C-terminus at the bottom of each chain.

[0093] Both the light and heavy chains are divided into regions of structural and functional homology. The terms “constant” and “variable” are used functionally. The variable domains of both the light (VL) and heavy (VH) chain portions determine antigen recognition and specificity. Conversely, the constant domains of the light chain (CL) and the heavy chain (CHI, CH2 or CH3) confer important biological properties such as secretion, transplacental mobility, Fc receptor binding, complement binding, and the like. The term "antigen-binding site," or "binding portion" can refer to the part of the immunoglobulin molecule that participates in antigen binding. The antigen binding site is formed by amino acid residues of the N-terminal variable ("V") regions of the heavy ("H") and light ("L") chains. Three highly divergent stretches within the V regions of the heavy and light chains, referred to as "hypervariable regions," are interposed between more conserved flanking stretches known as "framework regions," or "FRs". Thus, the term "FR" can refer to amino acid sequences which are naturally found between, and adjacent to, hypervariable regions in immunoglobulins. In an antibody molecule, the three hypervariable regions of a light chain and the three hypervariable regions of a heavy chain are disposed relativeto each other in three-dimensional space to form an antigen-binding surface. The antigen-binding surface is complementary to the three-dimensional surface of a bound antigen, and the three hypervariable regions of each of the heavy and light chains are referred to as "complementaritydetermining regions," or "CDRs."

[0094] The six CDRs present in each antigen-binding domain are short, non-contiguous sequences of amino acids that are specifically positioned to form the antigen-binding domain as the antibody assumes its three-dimensional configuration in an aqueous environment. The remainder of the amino acids in the antigen-binding domains, the FR regions, show less inter- molecular variability. The framework regions can adopt a P-sheet conformation and the CDRs form loops which connect, and in some cases form part of, the P-sheet structure. The framework regions act to form a scaffold that provides for positioning the CDRs in correct orientation by inter-chain, non-covalent interactions. The antigen-binding domain formed by the positioned CDRs provides a surface complementary to the epitope on the immunoreactive antigen, which promotes the non-covalent binding of the antibody to its cognate epitope. The amino acids comprising the CDRs and the framework regions, respectively, can be readily identified for a heavy or light chain variable region by one of ordinary skill in the art, since they have been previously defined (See, “Sequences of Proteins of Immunological Interest,” Kabat, E., et al., U.S. Department of Health and Human Services, (1983); and Chothia and Lesk, J. Mol. Biol., 196:901-917 (1987)).

[0095] Where there are two or more definitions of a term which is used and / or accepted within the art, the definition of the term as used herein is intended to include all such meanings unless explicitly stated to the contrary. The CDR definitions according to Kabat and Chothia include overlapping or subsets of amino acid residues when compared against each other. Nevertheless, application of either definition to refer to a CDR of an antibody or variants thereof is intended to be within the scope of the term as defined and used herein. The appropriate amino acid residues which encompass the CDRs as defined by each of the above cited references are set forth in the table below as a comparison. The exact residue numbers which encompass a particular CDR will vary depending on the sequence and size of the CDR. Those skilled in theart can routinely determine which residues comprise a particular CDR given the variable region amino acid sequence of the antibody.

[0096] Kabat et al. defined a numbering system for variable domain sequences that is applicable to any antibody. The skilled artisan can unambiguously assign this system of “Kabat numbering” to any variable domain sequence, without reliance on any experimental data beyond the sequence itself. As used herein, “Kabat numbering” can refer to the numbering system set forth by Kabat et al., U.S. Dept, of Health and Human Services, “Sequence of Proteins of Immunological Interest” (1983).

[0097] Antibodies may be polyclonal or monoclonal; xenogeneic, allogeneic, or syngeneic; or modified forms thereof (for example, humanized, chimeric, etc.). Antibodies may also be fully human. Preferably, antibodies of the invention bind specifically or substantially specifically to a component of cBAF complex. The terms “monoclonal antibodies” and “monoclonal antibody composition,” as used herein, refer to a population of antibody polypeptides that contain only one species of an antigen binding site capable of immunoreacting with a particular epitope of an antigen, whereas the term “polyclonal antibodies” and “polyclonal antibody composition” refer to a population of antibody polypeptides that contain multiple species of antigen binding sites capable of interacting with a particular antigen. A monoclonal antibody composition typically displays a single binding affinity for a particular antigen with which it immunoreacts.Methods - Immunization

[0098] The immune response against the compositions of the invention can be generated by one or more inoculations of a subject with an immunogenic composition of the invention. A first inoculation can be termed a “primary inoculation” and subsequent immunizations can be termed “booster inoculations”. Booster inoculations can enhance the immune response, and immunization regimens including at least one booster inoculation can be used. Any composition of the invention may be used for a primary or booster immunization. The adequacy of the vaccination parameters chosen, e g., formulation, dose, regimen and the like, can be determined by taking aliquots of serum from the subject and assaying antibody titers during the course of theimmunization program. Alternatively, T cell populations can by monitored by conventional methods. The clinical condition of a subject can be monitored for the desired effect, e.g., limiting Paramyxovirus and / or Henipavirus infection, improvement in disease state (e.g., reduction in viral load), etc. If such monitoring indicates that vaccination is sub-optimal, the subject can be boosted with an additional dose of composition, and the vaccination parameters can be modified in a fashion expected to potentiate the immune response. Thus, for example, the dose of the polypeptide, VLP, or composition, and / or adjuvant, can be increased or the route of administration can be changed.

[0099] As described herein, aspects of the invention are drawn to compositions and methods of preventing a viral infection in a subject. For example, use of the F or G proteins or portions thereof as immunogens to elicit an immune response to a Henipavirus.

[0100] For example, one or more of the following effects can result from the administration of a therapy or a combination of therapies as described herein: (i) the reduction or amelioration of the severity of a viral infection and / or a symptom associated therewith; (ii) the reduction in the duration of a viral infection and / or a symptom associated therewith; (iii) the regression of a viral infection and / or a symptom associated therewith; (iv) the reduction of the titer of a virus; (v) the reduction in organ reduced function or failure associated with a viral infection; (vi) the reduction in hospitalization of a subject; (vii) the reduction in hospitalization length; (viii) the increase in the survival of a subject; (ix) the elimination of a virus infection; (x) the inhibition of the progression of a viral infection and / or a symptom associated therewith; (xi) the prevention of the spread of a virus from a cell, tissue or subject to another cell, tissue or subject; and / or (xii) the enhancement or improvement the therapeutic effect of another therapy.

[0101] In embodiments, a therapeutically effective amount can comprise less than about 0.1 mg / kg, about 0.1 mg / kg, about 0.5 mg / kg, about 1.0 mg / kg, about 2.5 mg / kg, about 5 mg / kg, about 7.5 mg / kg, about 10 mg / kg, about 15 mg / kg, about 20 mg / kg, about 25 mg / kg, about 30 mg / kg, about 35 mg / kg, about 40 mg / kg, about 45 mg / kg, about 50 mg / kg, about 55 mg / kg, about 60 mg / kg, about 70 mg / kg, about 80 mg / kg, about 90 mg / kg, about 100 mg / kg, about 120 mg / kg, about 135 mg / kg, about 150 mg / kg, about 175 mg / kg, about 200 mg / kg, about 225mg / kg, about 250 mg / kg, about 275 mg / kg, about 300 mg / kg, about 325 mg / kg, about 350 mg / kg, about 375 mg / kg, about 400 mg / kg, about 425 mg / kg, about 450 mg / kg, about 475 mg / kg, about 500 mg / kg, about 525 mg / kg, about 550 mg / kg, about 575 mg / kg, about 600 mg / kg, about 625 mg / kg, about 650 mg / kg, about 675 mg / kg, about 700 mg / kg, about 725 mg / kg, about 750 mg / kg, about 775 mg / kg, about 800 mg / kg, about 825 mg / kg, about 850 mg / kg, about 875 mg / kg, about 900 mg / kg, about 1.0 g / kg, about 1.5 g / kg, about 2.0 g / kg, about 2.5 g / kg, about 5 g / kg, about 10 g / kg, about 25 g / kg, about 50 g / kg, or more than 50 g / kg of compound per body weight of a subject.

[0102] In embodiments, the therapeutically effective amount comprises less than about 0.1 mg, about 0.1 mg, about 0.5 mg, about 1.0 mg, about 2.5 mg, about 5 mg, about 7.5 mg, about 10 mg, about 15 mg, about 20 mg, about 25 mg, about 30 mg, about 35 mg, about 40 mg, about 45 mg, about 50 mg, about 55 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 120 mg, about 135 mg, about 150 mg, about 175 mg, about 200 mg, about 225 mg, about 250 mg, about 275 mg, about 300 mg, about 325 mg, about 350 mg, about 375 mg, about 400 mg, about 425 mg, about 450 mg, about 475 mg, about 500 mg, about 525 mg, about 550 mg, about 575 mg, about 600 mg, about 625 mg, about 650 mg, about 675 mg, about 700 mg, about 725 mg, about 750 mg, about 775 mg, about 800 mg, about 825 mg, about 850 mg, about 875 mg, about 900 mg, about 1.0 g, about 1.5 g, about 2.0 g, about 2.5 g, about 5 g, about 10 g, about 25 g, about 50 g, or more than 50 g.

[0103] Aspects of the invention are also drawn to managing a subject afflicted with or at risk of a viral infection.

[0104] Aspects of the invention can comprise administering to a subject an immunogenic composition comprising an epitope from a mutant Henipavirus fusion protein or G protein antigen, or combinations thereof.

[0105] In embodiments, administering can refer to providing a therapeutically effective amount of the composition to a subject. The formulation or pharmaceutical compound can be administered alone, but can be administered with other compounds, excipients, fillers, binders, carriers or other vehicles selected based upon the chosen route of administration and standardpharmaceutical practice. In some embodiments, an immune response can be stimulated in an individual by administering to the individual the immunogenic composition described herein or the amino acid epitope grafted into the scaffold protein described herein. In embodiments, the method is for eliciting an immune response to a fusion peptide epitope and / or a mutant Henipavirus fusion protein antigen.

[0106] Administration can be by way of carriers or vehicles, such as injectable solutions, including sterile aqueous or non-aqueous solutions, or saline solutions; creams; lotions; capsules; tablets; granules; pellets; powders; suspensions, emulsions, or microemulsions; patches; micelles; liposomes; vesicles; implants, including microimplants; eye drops; other proteins and peptides; synthetic polymers; microspheres; nanoparticles; and the like.

[0107] In embodiments, the compound can be administered alone, or can be administered as a pharmaceutical composition together with other compounds, excipients, carriers, diluents, fillers, binders, or other vehicles selected based upon the chosen route of administration and standard pharmaceutical practice. Administration can be by way of carriers or vehicles, such as injectable solutions, including sterile aqueous or non-aqueous solutions, or saline solutions; creams; lotions; capsules; tablets; granules; pellets; powders; suspensions, emulsions, or microemulsions; patches; micelles; liposomes; vesicles; implants, including microimplants; eye drops; other proteins and peptides; synthetic polymers; microspheres; nanoparticles; and the like.

[0108] Embodiments can be administered to a subject in one or more doses. The dose level can vary as a function of the specific composition or pharmaceutical composition administered, the severity of the symptoms and the susceptibility of the subject to side effects. Dosages for a given compound are readily determinable by a variety of means. For example, dosages can be determined by standard clinical techniques. In addition, in vitro or in vivo assays can be employed to help identify optimal dosage ranges. The precise dose to be employed can also depend on the route of administration and can be decided according to the judgment of the practitioner and each patient's circumstances.

[0109] In some embodiments, multiple doses of the composition can be administered. The frequency of administration and the duration of administration of the composition can varydepending on any of a variety of factors, e.g., patient response, severity of the symptoms, and the like. For example, in an embodiment, the pharmaceutical composition can be administered once per month, twice per month, three times per month, every other week (qow), once per week (qw), twice per week (biw), three times per week (tiw), four times per week, five times per week, six times per week, every other day (qod), daily (ad), twice a day (qid), three times a day (tid), or four times a day. In an embodiment, the pharmaceutical composition can be administered 1 to 4 times a day over a period of time, such as 1 to 10-day time period, or longer than a 10-day period of time.

[0110] In embodiments, the composition can be administered in combination with one or more additional active agents. For example, a first agent (e.g., a prophylactic or therapeutic agent) can be administered prior to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks before), concomitantly with, or subsequent to (e.g., 5 minutes, 15 minutes, 30 minutes, 45 minutes, 1 hour, 2 hours, 4 hours, 6 hours, 12 hours, 24 hours, 48 hours, 72 hours, 96 hours, 1 week, 2 weeks, 3 weeks, 4 weeks, 5 weeks, 6 weeks, 8 weeks, or 12 weeks after) the administration of a second agent (e.g., a prophylactic or therapeutic agent) to a subject with a disease or disorder or a symptom thereof.

[0111] Embodiments as described herein further comprises administering one or more additional active agents to a subject together with a mutant Henipavirus fusion protein antigen. Non-limiting examples of such additional active agents can comprise an anti-viral agent (e.g., remdesivir, molunpiravir, paxlovid, or any combination thereof), a vaccine, an anti-inflammatory agent, a pain reliever, a steroid, or any combination thereof.Kits

[0112] Aspects of the invention are also directed towards kits, such as kits comprising compositions as described herein. For example, the kit can comprise prophylactic and / or therapeutic combination compositions described herein.

[0113] In one embodiment, the kit includes (a) a container that contains a composition, such as that described herein, and optionally (b) informational material. The informational material can be descriptive, instructional, marketing or other material that relates to the methods described herein and / or the use of the agents for therapeutic benefit.

[0114] In an embodiment, the kit includes two or more agents. For example, the kit can include a container comprising an immunogenic composition comprising an epitope from mutant Henipavirus fusion protein antigen, and a second container comprising a second active agent.

[0115] In embodiments, the kit further comprises a third container comprising a third active agent.

[0116] The informational material of the kits is not limited in its form. In one embodiment, the informational material can include information about production of the compound, molecular weight of the compound, concentration, date of expiration, batch or production site information, and so forth. In one embodiment, the informational material relates to methods of administering the therapeutic combination composition, e.g., in a suitable dose, dosage form, or mode of administration (e.g., a dose, dosage form, or mode of administration described herein), to treat a subject who has a Henipavirus infection. The information can be provided in a variety of formats, include printed text, computer readable material, video recording, or audio recording, or information that provides a link or address to substantive material.

[0117] The composition in the kit can include other ingredients, such as a solvent or buffer, a stabilizer, or a preservative. The immunogenic composition can be provided in any form, e.g., liquid, dried or lyophilized form, or for example, substantially pure and / or sterile. When the agents are provided in a liquid solution, the liquid solution is an aqueous solution. When the agents are provided as a dried form, reconstitution generally is by the addition of a suitable solvent. The solvent, e.g., sterile water or buffer, can optionally be provided in the kit.

[0118] The kit can include one or more containers for the composition or compositions containing the agents. In some embodiments, the kit contains separate containers, dividers or compartments for the composition and informational material. For example, the composition can be contained in a bottle, vial, or syringe, and the informational material can be contained in aplastic sleeve or packet. In other embodiments, the separate elements of the kit are contained within a single, undivided container. For example, the composition is contained in a bottle, vial or syringe that has attached thereto the informational material in the form of a label. In some embodiments, the kit includes a plurality (e.g., a pack) of individual containers, each containing one or more unit dosage forms (e.g., a dosage form described herein) of the agents. The containers can include a combination unit dosage, e.g., in a desired ratio. For example, the kit includes a plurality of syringes, ampules, foil packets, blister packs, or medical devices, e.g., each containing a single combination unit dose. The containers of the kits can be airtight, waterproof (e.g., impermeable to changes in moisture or evaporation), and / or light-tight. The kit optionally includes a device suitable for administration of the composition, e.g., a syringe or other suitable delivery device. The device can be provided pre-loaded with one or both of the agents or can be empty, but suitable for loading.Sequences

[0119] Full-length F proteins from additional viruses are shown below (SEQ ID NOs; 70- 131). These proteins have a proline at the position equivalent to position 447 in AngV.

[0120] SEQ ID NO: 70, NiV-Ml (NC 002728)

[0121] MVVILDKRCYCNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNP LTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVY VLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDPVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0122] SEQ ID NO: 71, NiV-M2 (AF376747)

[0123] MVVILDKRCYCNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVY VLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPTTNNMRECLT GSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCP TAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDPVDISSQISSMNQSLQQSKDYIKEAQR LLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLYY IGT

[0124] SEQ ID NO: 72, NiV-M3 (AJ627196)

[0125] MVVILDKRCYCNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVY VLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYAIEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDPVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0126] SEQ ID NO: 73, NiV-M4 (FN869553)

[0127] MVVILDKRCYSNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIANVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQKSSMNQSLQQSKDYIKEA QRLLDTVNP SLISML S M 11 L YVLSIASLCIGLITFISFIIVEKKRNT YSRLEDRRVRPT S SGDL YYIGT

[0128] SEQ ID NO: 74, NiV-C (MK801755)

[0129] MVVVLDKRCYSNLLMLILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNP LTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVIKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAVGPPVFTDpVDISSQISSMNQSLQQSKDYIKEA QRLLDTVNP SLISML SMIIL YVLSIASLCIGLITFISFIIVEKKRNT YSRLEDRRVRPT S SGDLYYIGT

[0130] SEQ ID NO: 75, NiV-Bl .l (AY988601)

[0131] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPL TKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYSSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEGFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLYYIGT

[0132] SEQ ID NO: 76, NiV-B1.2 (MK673568)

[0133] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPL TKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0134] SEQ ID NO: 77, NiV-B1.3 (JN808864)

[0135] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLYYIGT

[0136] SEQ ID NO: 78, NiV-B1.4 (MK673584)

[0137] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSINYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0138] SEQ ID NO: 79, NiV-B2.3 (MK673579)

[0139] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKVGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVY VLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYTTEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYI QELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMREC LTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0140] SEQ ID NO: 80, NiV-B3.1 (MW535746)

[0141] MAVILNMRYYSNLLILILMISEC S VGILHYEKL SKIGL VKGITRKYKIKSNPLTK DIVIKMIPNVSNMSQCTGSVMENYKTRLNGILMPIKGALEIYKNNTHDLVGDVRLAGVI MAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLT ALQDYINTNLVPTIDKISCKQMELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQA FGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELL PVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGS TEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPT AVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLL DTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSNGDLYYIGT

[0142] SEQ ID NO: 81, NiV-B2.8 (PV132707)

[0143] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIVSLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNP SLISML SMIIL YVLSIASLCIGLITFISFIIVEKKRNT YSRLEDRRVRPT S SGDL YYIGT

[0144] SEQ ID NO: 82, NiV-B1.9 (PP981667)

[0145] MAVILNKGYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSESIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0146] SEQ ID NO: 83, NiV-B2.13 (PP981682)

[0147] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYPSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPT A VLGN VIISLGKYLGS VNYNSEGIAIGPP VFTDp VDI S SQI S SMNQ SLQQ SKD YIKEAQ RLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0148] SEQ ID NO: 84, NiV-I2.2 (OR820508)

[0149] MAVILNKRYYSNLLLLILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDLRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKD YIKEAQRLLDTVNPSLISMLSMIILYVLSIASLCIGLITFISFIIVEKKRNTYSRLEDRRVRPTSSGDLY YIGT

[0150] SEQ ID NO: 85, HeV-al .1 (NC 001906)

[0151] MATQEVRLKCLLCGIIVLVL SLEGLGILHYEKL SKIGL VKGITRKYKIKSNP LTKDIVIKMIPNVSNVSKCTGTVMENYKSRLTGILSPIKGAIELYNNNTHDLVGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSIAGQIVYVDLSSYYIIVRVYFPILTEIQQAYV QELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLITKKSVICNQDYATPMTASVREC LTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNT TCTTVVLGNIIISLGKYLGSINYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEAQKILDTVNPSLISMLSMIILYVLSIAALCIGLITFISFVIVEKKRGNYSRLDDRQVRPVSNG DLYYIGT

[0152] SEQ ID NO : 86, HeV-a 1.2 (HM044318)

[0153] MATQEVRLKCLLCGIIVLVL SLEGLGILHYEKL SKIGL VKGITRKYKIKSNP LTKDIVIKMIPNVSNVSKCTGTVMENYKSRLTGILSPIKGAIELYNNNTHDLVGDVKLAG VVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYV QELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLITKKSVICNQDYATPMTASVRECLTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNT TCTTVVLGNIIISLGKYLGSINYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEA QKILDTVNPSLISMLSMIILYVLSIAALCIGLITFISFVIVEKKRGNYSRLDDRQVRPVSNG DLYYIGT

[0154] SEQ ID NO: 87, HeV-a7 (PQ660692)

[0155] MATQEVRLKCLLCGIIVLVL SLEGLGILHYEKL SKIGL VKGITRKYKIKSNP LTKDIVIKMIPNVSNVSKCTGTVMENYKSRLTGILSPIKGAIELYNNNTHDLVGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDQISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYV QELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLITKKSVICNQDYATPMTASVREC LTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNT TCTTVVLGNIIISLGKYLGSINYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEAQKILETVNPSLISMLSMIILYVLSIAALCIGLITFISFVIVEKKRGNYSRLDDRQVRPVSNGD LYYIGT

[0156] SEQ ID NO: 88, HeV-pl (MZ318101)

[0157] MATQRVKVSYLICGIVISALSLEGLGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNVSKCTGTVMENYKNRLTGILSPIKGAIELYNNNTHDLIGDVKLAG VVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYV QELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLTTKKSVICNQDYATPMTSTVRECLTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCTTVVLGNIIISLGKYLGSTNYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEAQRILDTVNPSLISMLSMIVLYVLSTAALCIGLITLISFIIVEKKRGNYSRLDDRRVRPVSNGDLYYIGT

[0158] SEQ ID NO: 89, HeV-p2 (MZ229747)

[0159] MATQGVKVSYLICGIVISALSLEGLGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNVSKCTGTVMENYKNRLTGILSPIRGAIELYNNNTHDLIGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYVQELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLTTKKSVICNQDYATPMTSTVRECLTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCTTVVLGNIIISLGKYLGSTNYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEAQRILDTVNPSLISMLSMIVLYVLSTAALCIGLITLISFIIVEKKRGNYSRLDDRRVRPVSNGDLYYIGT

[0160] SEQ ID NO: 90, CedV-1 (JQ001776)

[0161] MSNKRTTVLIIISYTLFYLNNAAIVGFDFDKLNKIGVVQGRVLNYKIKGDPMTKDLVLKFIPNIVNITECVREPLSRYNETVRRLLLPIHNMLGLYLNNTNAKMTGLMIAGVIMGGIAIGIATAAQITAGFALYEAKKNTENIQKLTDSIMKTQDSIDKLTDSVGTSILILNKLQTYINNQLVPNLELLSCRQNKIEFDLMLTKYLVDLMTVIGPNINNPVNKDMTIQSLSLLFDGNYDIMMSELGYTPQDFLDLIESKSITGQIIYVDMENLYVVIRTYLPTLIEVPDAQIYEFNKITMSSNGGEYLSTIPNFILIRGNYMSNIDVATCYMTKASVICNQDYSLPMSQNLRSCYQGETEYCPVEAVIASHSPRFALTNGVIFANCINTICRCQDNGKTITQNINQFVSMIDNSTCNDVMVDKFTIKVGKYMGRKDINNINIQIGPQIIIDpVDLSNEINKMNQSLKDSIFYLREAKRILDSVNISLISPSVQLFLIIISVLSFIILLIIIVYLYCKSKHSYKYNKFIDDPDYYNDYKRERI NGKASKSNNIYYVGD

[0162] SEQ ID NO: 91, GhV(+A) (HQ660129)

[0163] MVINGNIITIILLLILILKTQMSEGAIHYETLSKIGLIKGITREYKVKGTP S SKDIVIKLIPNVTGLNKCTNISMENYKEQLDKILIPINNIIELYANSTKSAPGNARFAGVIIAGVA LGVAAAAQITAGIALHEARQNAERINLLKDSISATNNAVAELQEATGGIVNVITGMQDYI NTNLVPQIDKLQCSQIKTALDISLSQYYSEILTVFGPNLQNPVTTSMSIQAISQSFGGNIDL LLNLLGYTANDLLDLLESKSITGQITYINLEHYFMVIRVYYPIMTTISNAYVQELIKISFNV DGSEWVSLVPSYILIRNSYLSNIDISECLITKNSVICRHDFAMPMSYTLKECLTGDTEKCP REAVVTSYVPRFAISGGVIYANCLSTTCQCYQTGKVIAQDGSQTLMMIDNQTCSIVRIEEI LISTGKYLGSQEYNTMHVSVGNPVFTDpLDITSQISNINQSIEQSKFYLDKSKAILDKINLN LIGSVPISILFIIAILSLILSIITFVIVMIIVRRYNKYTPLINSDPSSRRSTIQDVYIIPNPGEHSIR SAARSIDRDRD

[0164] SEQ ID NO: 92, AngV (ON613535)

[0165] MGFRKNLRLLGIFWLIILSDTVDFERLASIGGIKGHSKLYKIKGHPTTKDIVI KLVPNLNNLTVCSEMSIDGYLNLINAVIIPISQSLELMRNNVKDGTPNENIFGAIVAGAAL GIATAAQVTSVVALHKSNQNAAKINSLRDSITKTNQAVEQLSLGVQETVSVLMGLQDQI NTNLVPKINILSCKQLSNTLNIMLLQYYSQILTVFGPNLRDPATVPVSIQALSQLFEGNLE LLTSSLGISSTDFNDLLTGKLITGSIIWADTQGHYLILRVNIPDLIEVPGAVIQEFIKIGYNF GGSLWMPTIPNKIIIRGYHLSLIDSTNCIITDNSYVCDRDYSLPMNPILRECFEGNTSSCGR EMVLNSYIPLYALSEGVIYANCLATSCKCATTNKPIVQSTSTVITMIDNSKCPVVEVGKQ MISVGSYLGQVSPNNLTIEIGPPVYTEPVDITNQLGNINETLSKTLDLIKQSNSILDMITGG LNPQIGSMISLILAAIAILLSGITCLFSTKSYVKCNKLQMNCFKSYERMDPIYHSQQN

[0166] SEQ ID NO: 93, LayV-Cl (OM101125)

[0167] MAFLKSAIICYLLFYPHIVKSSLHYDSLSKVGIIKGLTYNYKIKGSPSTKLM VVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNAMLDNVKSGNNKYRFAGAI MAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQNTNEAVKQLQLANKQTLAV IDTIRGEINNNUPVINQLSCDTIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISSVFN RNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQELM PISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIGCLQ GDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTVSLLDNKKCLVYQVGDILISVGSYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIEKS EEFLKGINPSIITLGSMAVLYIFMIVIAVISIIALVLSIKLTVKGNVVRQQFAYTQHVPSME NVNYVSH

[0168] SEQ ID NO : 94, Lay V-C2 (OM 101130)

[0169] MAFLKSAIICYLLFYPHIVKSSLHYDSLSKVGIIKGLTYNYKIKGSPSTKLMVVKLIPNIDRVRNCTQKQFDEYKNLVKNVLEPVKLALNAMLDNVKSGNNKYRFAGAIM AGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQNTNEAVKQLQLANKQTLAVID TIRGEINNNIIPVINQLSCDTIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISSVFNR NFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQELMPI SYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIGCLQGD TSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTVSLLDNKKCL VYQVGDILISVGSYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIEKSEE FLKGINPSIITLGSMAVLYIFMIVIA VISUAL VLSIKLTVKGNVVRQQFAYTQHVPSMENV NYVSH

[0170] SEQ ID NO: 95, LayV-K2 (PQ641429)

[0171] MGRVVSLKLTLIWYLLLYSCIVETSLHYDSLSKVGIIKGLTYNYKIKGSPSTKLMVVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNTMLDNVKSGNNKYRFA GAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQSTNEAVKQLQLANKQTL AVIDTIRGEINNNIIPVINQLSCETIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISS VFNRNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQ ELMPISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIG CLHGDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTISLLDN KRCLVYQVGDILISVGTYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIE KSEEFLKGINPSIITLGSMAVLYIFMIVIA VISUAL VLSIKLTVKGKVVRQQFAYTQHVPSM ENVNYVSH

[0172] SEQ ID NO: 96, LayV-K1.2 (PQ641427)

[0173] MERVVFLKLTLIW YLLLY SCTVETSLHYD SLSK VGIIKGLT YNYKIKGSP ST KLMVVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNTMLDNVKSGNNKYRFAGAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQSTNEAVKQLQLANKQTL AVIDTIRGEINNNIIPVINQLSCETIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISS VFNRNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQ ELMPISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIGCLQGDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTISLLDN KKCLVYQVGDILISVGTYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIE KSEEFLKGINPSIITLGSMAVLYIFMIVIA VISUAL VLSIKLTVKGKVVRQQFAYTQHVPSM ENVNYVSH

[0174] SEQ ID NO: 97, LayV-K1.3 (PQ641428)

[0175] MGRVVYLKLTLIW YLLL YSCI VETSLHYD SLSKVGIIKGLT YNYKIKGSP STKLMVVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNTMLDNVKSGNNKYRFA GAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQSTNEAVKQLQLANKQTL AVIDTIRGEINNNIIPVINQLSCETIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISS VFNRNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQ ELMPISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIG CLQGDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTISLLDN KRCLVYQVGDILISVGTYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIEKSEEFLKGINPSIITLGSMAVLYIFMIVIA VISUAL VLSIKLTVKGKVVRQQFAYTQHVPSM ENVNYVSH

[0176] SEQ ID NO: 98, MojV (NC 025352)

[0177] MALNKNMF S SLFLGYLL VYATT VQ S SIHYD SL SKVGVIKGLT YNYKIKGSP STKLMVVKLIPNIDSVKNCTQKQYDEYKNLVRKALEPVKMAIDTMLNNVKSGNNKYRF AGAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKSAIQNTNEAVKQLQLANKQ TLAVIDTIRGEINNNIIPVINQLSCDTIGLSVGIRLTQYYSEIITAFGPALQNPVNTRITIQAIS SVFNGNFDELLKIMGYTSGDLYEILHSELIRGNIIDVDVDAGYIALEIEFPNLTLVPNAVV QELMPISYNIDGDEWVTLVPRFVLTRTTLLSNIDTSRCTITDSSVICDNDYALPMSHELIG CLQGDTSKCAREKVVSSYVPKFALSDGLVYANCLNTICRCMDTDTPISQSLGATVSLLD NKRCSVYQVGDVLISVGSYLGDGEYNADNVELGPPIVIDpIDIGNQLAGINQTLQEAEDYIEKSEEFLKGVNPSIITLGSMVVLYIFMILIAIVSVIALVLSIKLTVKGNVVRQQFTYTQHV PSMENINYVSH

[0178] SEQ ID NO: 99, SHNV5-1 (OQ715593)

[0179] MINMD SKRLKVKVVFPIYLIIYIQMSRASLD YDQL SKIGIVKGP S YNYKIKGSPSTKLMVVKLVPNIKEVANCTHKQVENYKTLVRNVLDPVKMSLQAMLEKVKSGNNK YRFAGAIMAGVALGVATAATVTAGIALHRSSENAQAIARIKGAIQNTNEAVKQLQLAN KQMLAVIDTIRGEINENIIPIMNELSCETIGLNVGIKLTQYYSEIITAFGPALQNPIGSKITIQ Al S S AFNGNFDELLK SMGYT SNDL YE VLQ S GLIRGNIID VDPE VGYI ALEIEFPNLTL VPN AIIQELMPISFNSEGDEWVTLMPRFILTRTTLLSNIDITKCTVTDRSVICDNDYALPMSNQL IECLRGDTMKCTREKVMSSYIPKFALSDGVVYANCLNTICRCMDTDTPITQSLRSTVTLL DNKACLVYQIGDILISVGSYLGNTEYNTQNITLGPPIVIDpIDIGNQLAGINQSLQNAEDYI EKSDEFLKGINPSVITLGSMVVLYIFMILIAIISIIALVLSIRLTTKSNMQKAQFSYTRQAPS MDNINYVSR

[0180] SEQ ID NO: 100, SHNV5-2 (MZ328275)

[0181] MINMESKRLKVKVVF SIYFIIYIQMSRASLD YDQLSKIGIVKGP S YNYKIKGSPSTKLMVVKLVPNIKEVANCTHKQIENYKTLVRNVLDPVKMSLQAMLEKVKSGNNKY RFAGAIMAGVALGVATAATVTAGIALHRSSENAQAIARIKGAIQNTNEAVKQLQLGNK QMLAVIDTIRGEINENIIPIMNELSCETIGLNVGIKLTQYYSEIITAFGPALQNPIGSKITIQAI SSAFNGNFDELLKSMGYTSNDLYEVLQSGLIRGNIIDVDPEAGYIALEIEFPNLTLVPNAII QELMPISFNSEGDEWVTLMPRFILTRTTLLSNIDITKCTVTDRSVICDNDYALPMSNQLIE CLRGDTMKCTREKVMSSYIPKFALSDGVVYANCLNTICRCMDTDTPITQSLRSTVTLLD NKACLVYQIGDILISVGSYLGNTEYNTQNITLGPPIVIDpIDIGNQLAGINQSLQNAEDYIE KSEEFLKGINPSVITLGSMVVLYIFMILIAIISIIALVLSIRLTTKSNMQKAQFSYTRQAPSM DNINYVSR

[0182] SEQ ID NO: 101, HasV (OR713881)

[0183] MTI VNTVNMKL VLL VI YLIIS SE YVK ASLD YNELSK VGVIKGL S YNYKIRGS PSKKLMVVKLIPNIDTVENCSKTQLENYKILVRNALEPVKLSIKAMLDNVKSGNNKYRF AGAIMAGVALGVATAATVTAGIALHQSKENAQAIANIKSAIQNTNEAVKQLQLANKQTLAVIDTIRGEINDNIIPVINQLSCQTIGLNIGIKLTQYYSEILTAFGPAMQNPVNSRITIQAISSTFNGNFDELIKIMGYTSSDLYEVLHSGLIRGNIIDVDPEVGYIALEIEFPNLTLVPNAIIQELMPISFNIDGDEWVSLVPRFVLTRTTLLSNIDTDKCTVTEKSVICDNDYALPMSYQLVECLTGDTSKCTREKVISTYVPRFALSDGLIYANCLNTICRCMDTDTPIAQGLKSTVSLLDNKNCLVYQVGDILISVGTYLGETEYNTENIQLGPPVVIDpIDIGSQLAEINKTLQSAEDYIEKS DEFLKGVNP SVLTIGSMVIL YLFMIIIA VISUAL VLSIKLTVK SNVMKGQF S YVQHIP SMENVNYVSR

[0184] SEQ ID NO: 102, MelV (OK623353)

[0185] MAKFVFLKTMCIGLLIIISDRVDSSMDFHSLSKIGIIKGKTYNYKIRGEPNTKLMVIKLIPNIDVVENCSTTQVNNYKKLVRNVLTPVRIISRYYVKNVIEQNNRVRLFGAIMAGAALGVATAATVTAGIALHRSNENARNIALMKEAIKNTNQAVTKLQLAGQQTLAVIDNIRGEINNQIIPVINKLTCENIGLNVGIKLTQYYSEVLTAFGPAIQDPVNARITIQAISKVFNNNFDELLNVMGYSTQDLYEVLHGGLIRGNVISADPEVGYLALEIEFPNLSVVPNAYIQEIMPISFNVDGDEWVTVVPRHTLIRTTLLSNIDITTCSIIESSIICNNDYALPMSNELINCLQGS TDLCAREKVISNYVPKFALSDGVVYANCLSTVCRCMDNGVPISQSLKSTVMMLDDKKC TIYQIGDILISVGKYMGHIDYNPENVVLGPPIVIDpIDIGNQLAGINQTLQEAGDFIEKSEEI LNSINP S VLTL S SMITL YIFLIIT VIIAVAAL VLSIRLTIKARILTNQF A YGRHSP SMDNVS YVSK

[0186] SEQ ID NO: 103, DewV-1 (OK623354)

[0187] MARLQVVILYLYLLVASDVVKASLDFDNMSKIGIIKGNTYNYKIRGEPTTKLMVVKLIPNIDVVENCSATQLANYKKLVTNVLTPVKLSLDNMLKNVQDQNNRVRLFGAIMAGAALGVATAATVTAGIALHRSNENAKNIAKLKNAIQNTNLAITKLQMAGQQTLAVI DNIRGEINNQIIPVLNQLSCETVGLNVGIKLTQYYSEVLTAFGPAIQDPVNSKITIQAISKAFNNNFDELLKVMGYTSQDLYEILHGDLITGNVISADPEAGYIALEIEFPNLSTVPNAYVQE LMPISFNVDGDEW VTLVPRYVLIRTTFL SNIDISLC SIMET SIVCNND YALPMS SELINCLQ GDTGVCAREKVISSYVPKFALSGGVVYANCLSTVCRCMDNGTPISQGIRHTVALLDNKK CSVYQVGDILISVGKYLGHLDYNTEDIVLGPPIVIDpIDIGNQLAGINSSLQQAENYIDKSNEILKSINPTIISTNSMITLYIFIIITVIIAITSLVLSLRLTMKAKILTNQFAYGRHSPSMDNVSY VSR

[0188] SEQ ID NO: 104, DewV-2.1 (OR713882)

[0189] MAKLQMTIFYLYLLVASDLVETSLDFDNMSKIGIIKGNTYNYKIRGEPTTK LMVVKLIPNIDVVENCSATQLANYKKLVNNVLTPVKLSLDNMLKNVQDQNNRVRLFG AIMAGAALGVATAATVTAGIALHRSNENAKNIAKLKNAIQNTNLAITKLQMAGQQTLA VIDNIRGEINNQIIPVLNQLSCETVGLNVGIKLTQYYSEVLTAFGPAIQDPVNSKITIQAISK AFNNNFDELLKVMGYTSQDLYEILHGDLITGNVISADPETGYIALEIEFPNLSTVPNAYVQ ELMPISFNVDGDEWVTLVPRYVLIRTTFLSNIDISLCSIMETSIVCNNDYALPMSSELINCL QGDTGVCAREKVISSYVPKFALSGGVVYANCLSTVCRCMDNGTPISQGIKHTVALLDNK KCTVYQVGDILISVGKYLGHLDYNTEDIVLGPPIVIDpIDIGNQLAGINSSLQQAEDYIDKS NEILKGINPTIISTNSMITLYIFIIITVIIAITSLVLSLRLTMKAKILTNQFAYGRHSPSMDNVS YVSR

[0190] SEQ ID NO: 105, DewV-2.2 (OR713883)

[0191] M A K LQM T 1 F YL YLL VASDL VET SLDFDNMSKIGIIKGNTYNYKIRGEPTTK LMVVKLIPNIDVVENCSATQLANYKKLVNNVLTPVKLSLDNMLKNVQDQNNRVRLFG AIMAGAALGVATAATVTAGIALHRSNENAKNIAKLKNAIQNTNLAITKLQMAGQQTLA VIDNIRGEINNQIIPVLNQLSCETVGLNVGIKLTQYYSEVLTAFGPAIQDPVNSKITIQAISK AFNNNFDELLKVMGYTSQDLYEILHGDLITGNVISADPEAGYIALEIEFPNLSTVPNAYV QELMPISFNVDGDEWVTL VPRYVLIRTTFL SNIDISLC SVMET SIVCNNDYALPMS SELIN CLQGDTGVCAREKVISSYVPKFALSGGVVYANCLSTVCRCMDNGTPISQGIKHTVALLD NKKCTVYQVGDILISVGKYLGHLDYNTEDIVLGPPIVIDpIDIGNQLAGINSSLQQAEDYI DKSNEILKGINPTIISTNSMITLYIFIIITVIIAITSLVLSLRLTMKAKILTNQFAYGRHSPSMD NVSYVSR

[0192] SEQ ID NO: 106, DarV-K (MZ 574409)

[0193] MSSSNKIRITVIIINILISNYLIHCSMDFVELSRVGIIKGSTYNYKIRGEPSTKL MVVKLIPNVGDVANCSATQVSNYKKLVKNVLSPVSNALNTMLENVQTENNRYRLFGAI MAGAALGVATAATVTAGIALHRSNENARNIAAMKNAIQNTNQAITKLQMAGQQTLAVIDNIRGEINNQIIPLLNKLSCETVGLNVGIKLTQYYSEILTVFGPAIQDPINSKITIQAISKAFG GNFDELLKVMGYTSQDLYEILHGGLITGNIIGVDPDTGYIALEIEFPNLSIVQNAYIQELM PISFNVDGDEWVTLAPRFVLIRTTLLSNIDTSMCTIIDSSVICNNDYALPMSTELINCLQG MTESCAREKVISSYVPKFALSGGVIYANCLSTVCRCMDNQKPISQSLRSTVVMLDNKMCKVYQIGDILISVGEYKGAVEYNPEDVHLGPPIVLDpIDIGNQLAGINQTLSEAGDFISKSEE ILKDINPAILSVSSMIVLYIFIIVITIISLTSLILSIRLTIKAKIFNNQFAYGRHSPSMDNVSYVT H

[0194] SEQ ID NO: 107, DarV-C (OM030315)

[0195] MSSSNKIRITVIIINILISNYLINCSMDFVELSRVGIIKGSTYNYKIRGEPSTKLMVVKLVPNVGDVANCSTTQVSNYKKLVKNVLSPVSNALNTMLENVQTENNRYRLFGA IMAGAALGVATAATVTAGIALHRSNENARNIAAMKNAIQNTNQAITKLQMAGQQTLAV IDNIRGEINNQIIPLLNKLSCETVGLNVGIKLTQYYSEILTVFGPAIQDPINSKITIQAISKAF GGNFDELLNVMGYTSQDLYEILHGGLITGNIIGVDPDTGYIALEIEFPNLSIVQNAYIQELMPISFNVDGDEWVTLAPRFVLTRTTLLSNIDTSMCTIIDSSVICNNDYALPMSTELINCLQ GMTESCAREKVISSYVPKFALSGGVIYANCLSTVCRCMDNQKPISQSLRSTVVMLDNKM CKVYQIGDILISVGEYKGAVEYNPEDVHLGPPIVLDpIDIGNQLAGINQTLSEAGDFISKSE EILKDINP AIL S VS SMIVL YIFIIVITIISLTSLIL SIRLTIK AKIFNNQF A YGRHSP SMDNVS YVTH

[0196] SEQ ID NO: 108, DarV-Ca (PP272750)

[0197] MSSSNKIRITVIIINILISNYLINCSMDFVELSRVGIIKGSTYNYKIRGEPSTKLMVVKLIPNVGDVANCSATQVSNYKKLVKNVLSPVSNALNTMLENVQTENNRYRLFGAI MAGAALGVATAATVTAGIALHRSNENARNIAAMKNAIQNTNQAITKLQMAGQQTLAVI DNIRGEINNQIIPLLNKLSCETVGLNVGIKLTQYYSEILTVFGPAIQDPINSKITIQAISKAFG GNFDELLKVMGYTSQDLYEILHGGLITGNIIGVDPDTGYIALEIEFPNLSIVQNAYIQELMPISFNVDGDEWVTLAPRFVLTRTTLLSNIDTSMCTIIDSSVICNNDYALPMSTELINCLQG MTESCAREKVISSYVPKFALSGGVIYANCLSTVCRCMDNQKPISQSLRSTVVMLDNKMC KVYQIGDILISVGEYKGAVEYNPEDVHLGPPIVLDpIDIGNQLAGINQTLSEAGDFISKSEEILKDINPAILSVSSMIVLYIFIIVITIISLASLILSIRLTIKAKIFNNQFAYGRHSPSMDNVSYV TH

[0198] SEQ ID NO: 109, GakV-1 (MZ574407)

[0199] MELIKLFNIL YLVGYSGLFFTD VNAGLD YEGL S SIGVIKGPS YNYKIRGTP S TKLLVIKLIPNVESIDNCTQKQMSDYRALVKNVLTPVSESLSTMLNYIEQQSNGVRLIGAVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKMANQELLGV VDSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYSGSDLHDILQGDLITGNIIGVDPDVGYIALEINFPTLTEIPNAVIQ ELMPISFNDKGDEWMALVPRYVLLRTTYISNIDISKCLLTERSVICYNDYATPMSFDIIRC LTGNLT YCPREQIM ASHVPKF AL SGGVIYANCL S A VCRC AVDGVPIVQ SLK AT VMMLD NKSCRVYQIGELLISTGAYLGSIEFKNENIELGPPIVIDpIDLGGQIAGINQTLQGVEDYIDK SNEILDQVNPSVTSLGAMIIVYIFIALAILLGLIALIMDVKLNSQVKILINQSMINQMRAGLENPGYSRSLPSFGSIASRGSSQELVNTN

[0200] SEQ ID NO: 110, GakV-2 (MZ574408)

[0201] MELIKLFYILYLVGYSGLFFTDVNAGLDYEGLSSIGVIKGPSYNYKIRGTPSTKLLVIKLIPNVESIDNCTQKQMSDYRALVKNVLTPVSESLSTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKMANQELLGV VDSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYSGSDLHDILQGDLITGNIIGVDPDVGYIALEINFPTLTEIPNAVIQ ELMPISFNDKGDEWMALVPRYVLLRTTYISNIDISKCLLTERSVICYNDYATPMSFDIIRC LTGNLT YCPREQIM ASHVPKF AL SGGVIYANCL SA VCRC AVDGVPIVQ SLK AT VMMLD NKNCRVYQIGELLISTGAYLGSIEFKNENIELGPPIVIDpIDLGGQIAGINQTLQGVEDYIDKSNDILDQVNPSVTSLGAMIIVYIFIALAILLGLIALIMDVKLNSQVKILINQSMINQMRAGL ENPGYSRSLPSFGSVASRGSSQELVNTN

[0202] SEQ ID NO : 111 , SHNV 1 (OQ236120)

[0203] MNYTIVGIMIITFVHESQCINYEQLASIGVIKGHTYNYKIRGPPNTKLMVVKLIPNINIDNLGGGLSNCSSKQMESHKELVEKVLSPVAQALETMRNRVTDYSGNYRFVGA VMAGAALGVATAATVTAGIALHQSNQNAKAIDQMKEAIRTTNKAVQELTLSTRQTLLVIDSLQNQINTQIVPAMNRLSCEVLGLTVGIQLTQYYSEILTYFGPALQDPIDSTLTIQAISH AFGGNFDILMKTMGYTVGDLYDVLKGDLITGKIISVNPKEGFIALEVRFPTLTQVNNAIV QELMPISFNDKGDEWISTVPRYVLERVLYLSNIDISLCSVGETSVVCDNDYASPMSHQLR ECLQTNTSYCPRERVLASYVPKFALSQGVIFANCIATTCRCADDGRAISQSSSQTVLLLTS KDCKVYEVQSMMISTGEYLGESIFENTDIPLGPSIVIDpIDISGQLAEINKTLDHVDNTIKD SNDILDKIDVSTVSAISMVILYVVIALIGFMSALSLLLSVRTFSRCTTLGSQFAYQRQDPTL GD VHYAFT SNIPKNQRSKN SNNL SNDLGSTNGSF S SNE

[0204] SEQ ID NO : 112, SHNV2 (OM030316)

[0205] M SLKFRQSTNN RTN I RLI LI I VLIKHM EC IH YEN LS SVGIIKGNTYNYKIKGD PSTKMLVVKLIPNIDGLGNCTDKQMKDYKTLVKSTLQPVKDSLAQMLNNVETYNGYV KFFGAVMAGAALGVATAATVTAGIALHQSNQNAKDIANMKDAIMKTNQAITTLSSASQ KMLTVIDSIRGEINQQIIPVLNQLSCETVGLNLGIKLTQYYSQISTFFGPALQNPVQSILTIQ AISHAFGGNFNELMTVMGYTGSDFQDIIQGNLITGSVIGVDPEVGYIALEIRIPSLTVVPN AYVQELLPVSFNIDGDEWMTIVPNYVLTRTTYLSNIDINRCLITDKSVICGNDYATPMSN QLINCLNGNTQHCAREAVVTSYVPKFALSGGVIYANCLSTVCRCIDKDQPISQSLSQTLM MLDNQHCNVYQISNVLISTGRYLGDAEFRNEGIDLGPPIVVDpIDLGGQIADTNQTISDAE EFIEESNKILSKINPKIISVKSMVVLYVVVALIAILAMVSLVLSIRLTMQVGTKINQFAYTK HYP SMENVQ YIGTR

[0206] SEQ ID NO : 113 , SHNV3 (OM030317)

[0207] MELIKNYCLIYLIFYSSTLIRPLQAGLDYEALASIGVIKGPSYNYKIRGTPSTKLLVIKLIPNVGSLDNCTQKQMSDYRALVKNVLTPVSESLKTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDSIKNTNMAVQTLKLANQEILGV VDSLRGQINTQIIPVLNQLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDVGYIALEIHFPTLTEIPNAVIQ ELMPISFNDKGDEWMALVPRYVLLRTTYVSNIDISKCLLTERSVICYNDYATPMSFDVLR CLTGNLTFCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVDGVPIVQSLKTTIMMLDN KNCRVYQIGELLISTGAYLGSVEFKNENIELGPPIVVDpIDLGGQIAGINQTLQGVEDYIDKSNEILDQVNPSVTSLGAMIIVYIFIALAILIGLIALMIDVKLNSQIKILINQSIINQMRAGLE NPGYSKSLPSFSSTVSKGSSQELINTH

[0208] SEQ ID NO : 114, SHNV4 (OM030314)

[0209] MGLTKSTILVYLLVYYHAHIIAINAGLDYEGLASIGVVKGPSYNYKIRGTPTTKLLVIKLIPNVGSLDNCTQKQMADYKSLVKNVLTPVSDALSTMLNYIEQQSNGVRLIG AVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKLANQEILG VVDSLRGQINTQIIPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQE LMPISFNDKGDEWMTLVPRYVLLRTTYVSNIDISKCLITERSVICYNDYATPMSFDVIRCL TGNLTYCPREQIIASYVPRFALSGGVIYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKN CRVYQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQGVEDYIDKSN EILDQVNPSVTSLGAMIIVYIFIALAILIGLIALMIDVKLNSQIKILINQSIVNQMRAGLENP GYSRSLPSFSSIASKGSSQELVNTN

[0210] SEQ ID NO: 115, SHNV6 (PQ541138)

[0211] MAKESTSICISYLLIYCHLISGNLDYEALSKIGIIKGPSYNYKIKGTPSTKLMVVKLIPNL SNVENC SRGQIENYRNL VKNVLDP VKN ALD AMLENVKTGNNR YRF AGAIN! AGVALGVATAATVTAGIALHRSNENAQAIANMKSAIQSTNEAVKQLQTANQQMLAVID TVRGEINNNIIPVINQLSCETIGLNVGIKLTQYYSEVITAFGPALQNPVNTRITIQAISSAFN GNFDELLKIMGYTSNDLYEILQSGLIRGNIIDVDPIVGYIALEIEFPNLSLVPNAIIQELMPIS FNVDGDEWVSMVPYYVLTRTTLLSNIDINRCSVTTKSIICDNDYALPMSNQLIDCLKGET EKCARERVISSYVPKFALTDGVVYANCLHTICRCMDTDTPISQGLSSTVMLLDSKKCLV YQVGDILISVGEYLGESEYRTDNIELGPPIVIDpIDIGNQLAGINQTLQNAGDYIEKSEEFLS GVNPAVINLGSMIVLYIFIIIIAIISIIALVLSIKATSRGKFLKGQFAYSQHTPSMDSVSYVSH

[0212] SEQ ID NO: 116, SHNV7 (PQ541139)

[0213] MELVVIKKVIIIYILMYNHLIEASLDFNNLAKIGIIKGKTYNYKIRGEPNTKLMVVKLIPNIDVVQNCSGTQIANYKKLVNNVLSPIKLALDNMLKNVIEQNNRVRLFGAIM AGAALGVATAATVTAGIALHRSNENARNIALLKDSIKNTNQAVAKLQMANQQTLAVID NIRGEINNQIIPVMNKLTCETIGLNVGIKLTQYYSEILTVFGPAIQDPVNSKITIQAISKVFNNNFDELLNVMGYSSQDLYEILHGGLIR.GNVISADPEIGYMALEIEFPNLAVVPNAYIQEIM PISFNVDGDEWVTVVPRYTLIRTTLLSNIDISLCTLIETSVICNNDYALPMSSELVNCLQGN TEVCAREKVISNYVPKFALSDGVVYANCLSTVCRCMDNGVPISQNLKSTVMMLDDKKC TIYQIGDILISVGKYMGHVEYNPEDVVLGPPIVIDpIDIGNQLAGINQTLQEAGDFIEKSEEI LNSINPNVLSLSSMITLYIFLIITIIIAIAALVLSIRLTIKAKILTNQFAYGRHSPSMDNVSYVS K

[0214] SEQ ID NO: 117, SHNV8 (PP272530)

[0215] MKLTKYYCMLYLIGYAGFHFTEVDAGLDYEGLASIGVIKGPSYNYKIRGTPSTKLLVIKLIPNVDSIDNCTQKQMSDYKALVKNVLTPVSESLSTMLNYIEQQSNGVRLI GAVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKMANQELL GVVDSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAI SGAFSGNFDEMMRVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEINFPTLTEIPNAII QELMPISFNDKGDEWMALVPRFVLLRTTYISNIDISKCLLTERSVICYNDYATPMSFDIIR CLTGNLTYCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVDGVPIVQSLKATVMMLD NKNCRVYQIGELLISTGTYLGSVEFRNENIELGPPIVVDpIDLGGQIAGINQTLQGVEDYID KSNEILDQVNPSVTSLGAMIIVYIFIALAILLGLIALIIDVKLNSQIKILINQSMINQMRAGLE NPGYSRSLPSFGSVASRGSSQELVDTS

[0216] SEQ ID NO: 118, SHNV9 (PP272531)

[0217] MKLIRCICLLYLIGYSGLLLVNVDAGLDYEGLASIGVIKGPSYNYKIRGTPSTK LLVIKLIPNVGSIDNCTQKQMSDYKALVKNVLTPVAESLSTMLNYIEQQSNGVRLIGAVL AGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIRNTNQAVQTLKMANQELLGVV DSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAF SGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDVGYIALEINFPTLTEIPNAIIQEL MPISFNDKGDEWMALVPRYVLLRTTYISNIDTSKCLLTERSVICYNDYATPMSFDIIRCLT GNLTYCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVDGVPIVQSLKATVMMLDNKS CRVYQIGELLISTGTYLGSVEFRNENIELGPPIVVDpIDLGGQIAGINQTLQGVEDYIDKSN EILEQVNPSVTSLGAMIIVYIFIALAILLGLIALIIDVKLNSQIKILINQSMINQMRAGLENPGYSRSLPSFGSVASRGSSQELVDTS

[0218] SEQ ID NO: 119, SHNV10-1.2 (PP272749)

[0219] MELIKSTILVYLLIHQQLNIITVSAGLDYEGLASIGVVKGPSYNYKIRGTPTTKLLVIKLIPNVGSLDNCTQKQMADYKALVKNVLTPVSDALSTMLNYIEQQSNGVRLIGAVL AGAALGVATGAAITAGIALHKSNQNAQAIAQLRDAIKNTNQAVQTLKLANQELLGVVD SLRGQINTQIIPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAFSG NFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQELMPI SFNDKGDEWMTLVPRYVLLRTTYISNIDISKCLVTERSVICYNDYATPMSFDVIRCLTGNLTYCPREQIIASYVPRFALSGGVIYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKNCRV YQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQNVEDYIDKSNEILDQVNPSVTSLGAMIIVYIFIALAILIGLIALMIDIKLNSQIKILINQSIVNQMRAGLENPGYS RSLPSF S S VASKGS SQELVNTT

[0220] SEQ ID NO: 120, SHNV10-2.1 (PP272534)

[0221] MELIKSTILIYLLIHQQLNIITVSAGLDYEGLASIGVVKGPSYNYKIRGTPTTKLLVIKLIPNVGSLDNCTQKQMADYKALVKNVLTPVSDALSTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLRDAIKNTNQAVQTLKLANQELLGV VDSLRGQINTQIIPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAF SGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQELMPISFNDKGDEWMTLVPRYVLLRTTYISNIDISKCLVTERSVICYNDYATPMSFDVIRCLT GNLTYCPREQIIASYVPRFALSGGVVYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKN CRVYQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQGVEDYIDKSN GILDQ VNP S VT SLGAMIIVYIFI ALAILIGLIALMIDIKLNSQIKILINQ SIVNQMRAGLENPG YSRSLPSF SS VASKGS SQELVNTT

[0222] SEQ ID NO: 121, SHNV10-2.2 (PP272638)

[0223] MELIKSTILIYLLIHQQLNIITVSAGLDYEGLASIGVVKGPSYNYKIRGTPTTKLLVIKLIPNVGSLDNCTQKQMADYKALVKNVLTPVSDALSTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLRDAIKNTNQAVQTLKLANQELLGV VDSLRGQINTQIIPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAF SGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQELMPISFNDKGDEWMTLVPRYVLLRTTYISNIDISKCLVTERSVICYNDYATPMSFDVIRCLT GNLTYCPREQIIASYVPRFALSGGVVYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKN CRVYQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQGVEDYIDKSN EILDQVNPSVTSLGAMIIVYIFIALAILIGLIALMIDIKLNSQIKILINQSIVNQMRAGLENPG YSRSLPSF SS VASKGS SQELVNTT

[0224] SEQ ID NO : 122, SHNV 11 (OQ970176)

[0225] M K LN NT R RD N K I N KQHTTM ACT K NCC L V Y LI L YS STLIIPLEAGLD YESL ASIGVIKGPSYNYKIRGTPSTKLLVIKLIPNVGSLDNCTQKQMADYKALVKNVLTPVSESLK TMLNYIEQQSNGVRLIGAVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDSIKNT NMAVQTLKLANQEILGVVDSLRGQINTQIIPVLNQLSCDTVGLTLGIKLTQYYSEILTAFG PAIQDPVNSKLTIQAISGAFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDVGY IALEIHFPTLTEIPNAVIQELMPISFNDKGDEWMALVPRYVLLRTTYVSNIDISKCLLTERS VICYNDYATPMSFDVLRCLTGNLTFCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVD GVPIVQSLKTTIMMLDNKNCRVYQIGELLISTGSYLGSVEFKNENIELGPPIVVDpIDLGG QIAGINQTLQGVEDYIDKSNEILDQVNPSVTSLGAMIIVYIFIALAILIGLIALMIDVKLNSQ IKILINQSIINQMRAGLENPGYSRSLPSFSSTVSKGSSQELINTH

[0226] SEQ ID NO: 123, ChV-1 (PQ140950)

[0227] MARWLRNFIILYLIVFYSEYSGALHYENLSYVGVII<GLTYNYKII<GDPSTI< LLVIKLIPNLNITKENFNDCTRKQMDEHTKLVRSVLEPVKAALDAMRNKVTDYSGNYRF FGAVLAGAAMGVATAATITAGIALHKANENAYAINQMKDAIKNTNKAIQEVSNAQRQT VVVLDNLQGQINTQIIPVLNQLTCEMQGLTVGLQLTRYYSEILTFFGPAIQDPVNSVLTIQ AISHAFDRNFDALMTTMGYTAADMYEVLNSDLITGRIISVDPTIGYIALEIRFPTLTTIENA IIQELMPISFNHKSDEWITIVPKYVLERMSFISNIDTSLCLVGDKSIICDNDYATPMSTQMR NCLEGNTESCVREKVLASYVPKFALSGGVIYANCISAACRCADNGEAISQSSSSSVMMLS NQGCSTYEVQNTLISVGKYMGEKEFNSMDIEVGPPIVLDpVDISGQLTEINKTLDKAEEYI EESNDILKGIQVSLVSYTSMIVLYIVVALIGMMSVISLLLAVRLTSQAKILIRDMNYSRIDP TMYGVKYGFETPSTNSTIVSRNNNNNKLNGSVNGSFTSDE

[0228] SEQ ID NO : 124, ChV-2.1 (PQ 140949)

[0229] MARWSRNFIILYMILFYSEYSGALHYENLSYVGVIKGLTYNYKIKGDPSTKLLVIKLIPNLNITKENFNDCTRKQMDEHTKLVRSVLEPVKAALDAMRNKVTDYSGNYRF FGAVLAGAAMGVATAATITAGIALHKANENAYAINQMKDAIKNTNKAIQEVSNAQRQT VVVLDNLQGQINTQIIPVLNQLTCEMQGLTVGLQLTRYYSEILTFFGPAIQDPVNSVLTIQ AISHAFDRNFDALMTTMGYTAADMYEVLNSDLITGRIISVDPTIGYIALEIRFPTLTTIENA IIQELMPISFNHKSDEWITIVPKYVLERMSFISNIDTSLCLVGDKSIICDNDYATPMSTQMR NCLEGNTESCVREKVLASYVPKFALSGGVIYANCISAACRCADNGEAISQSSSSSVMMLS NQGCSTYEVQNTLISVGKYMGEKEFNSMDIEVGPPIVLDpVDISGQLTEINKTLDKAEDY IEESNDILKGIQVSLVSYTSMIVLYIVVALIGMMSVISLLLAVRLTSQAKILIRDMNYSRIDPTMYGVKYGFETPSTNSTIVSRNNNNNKLNGSVNGSFTSDE

[0230] SEQ ID NO: 125, ChV-2.2 (PQ140951)

[0231] MARWLRNFIILYMILFYSEYSGALHYENLSYVGVIKGLTYNYKIKGDPSTKLLVIKLIPNLNITKENFNDCTQKQMDEHTKLVRSVLEPVKAALDAMRNKVTDYSGNYRF FGAVLAGAAMGVATAATITAGIALHKANENAYAINQMKDAIKNTNKAIQEVSNAQRQT VVVLDNLQGQINTQIIPVLNQLTCEMQGLTVGLQLTRYYSEILTFFGPAIQDPVNSVLTIQ AISHAFDRNFDALMTTMGYTAADMYEVLNSDLITGRIISVDPTIGYIALEIRFPTLTTIENA IIQELMPISFNHKSDEWITIVPKYVLERMSFISNIDTSLCLVGDKSIICDNDYATPMSTQMR NCLEGNTESCVREKVLASYVPKFALSGGVIYANCISAACRCADNGEAISQSSSSSVMMLS NQGCSTYEVQNTLISVGKYMGEKEFNSMDIEVGPPIVLDpVDISGQLTEINKTLDKAEEYI EESNDILKGIQVSLVSYTSMIVLYIVVALIGMMSVISLLLAVRLTSQAKILIRDMNYSRIDPTMYGVKYGFETPSTNSTIVSRNNNNNKLNGSVNGSFTSDE

[0232] SEQ ID NO : 126, ResV-a (OR713876)

[0233] MKFLIIKYVTINYMLIYVSIFVKVSRSNLDYEALSSIGVIKGPSYNYKIRGNPITKLLVIKLIPNVGNISDCTTKQIQDYKTLVRNVLTPVSEALTTMLNYIEVQDNGVRFIGA VLAGAALGVATGAAITAGIALHKSNQNAQAISQMKDAIKNTNEAIQSLKMANQEMLSV VDSLRGQINTQIIPVMNQLGCQNIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPEVGYIALEIHFPTLTEIPNAVV QELMPISFNDKGDEWMTILPRYVLLRTTYTSNIDISKCLITDRSVICYNDYATPMSFDTIKCLTGNLTSCARERIIASHVPKFALSGGVIYANCLSVTCRCAIDGIPIVQSLRTTIMMLDNK KCRVYQIGELLISTGAYLGSIEFKNEDIELGPPIVLDpIDLGGQIAGINQTLQGVEDYIDKS NDILDKINPNVTSIGAMILVYIFIALSIVLGLSALIMGIKLNSQIKLLINQSIINQMRSGFDNA GYSRSLPSFSSTKSVNSTQELINTN

[0234] SEQ ID NO : 127, Res V- 1 (OR713877)

[0235] MRFVIIKYVTINYMLIYVSVFVKVSRSNLDYEALSSIGVIKRASYNYKIRGNPITKLLVIKLIPNVGNISDCTTKQIQDYKTLVRNVLTPVSEALTTMLNYIEVQDNGVRFIG AVLAGAALGVATGAAITAGIALHKSNQNAQAISQMKDAIKNTNEAIQSLKMANQEMLS VVDSLRGQINTQIIPVMNQLGCQNIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAIS GAFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPEVGYIALEIHFPTLTEIPNAV VQELMPISFNDKGDEWMTILPRYVLLRTTYTSNIDISKCLITDRSVICYNDYATPMSFDTI KCLTGNLTSCARERIIASHVPKFALSGGVIYANCLSVTCRCAIDGIPIVQSLRTTIMMLDN KKCRVYQIGELLISTGAYLGSIEFQNEDIELGPPIVLDpIDLGGQIAGINQTLQGVEDYIDK SNEILDKINPNVTSIGAMILVYIFIALSIVLGLSALIMGIKLNSQIKLLISQSIINQMRSGFDN AGYSRSLP SF S S TK S VNS TQELINTN

[0236] SEQ ID NO: 128, ResV- 2 (OR713878)

[0237] MRFFITRYVIINYMLIYISFFVKVGRSNLDYEALSSIGVIKGASYNYKIRGNPITKLLVIKLIPNVGNISDCTTKQIQDYKTLVRNVLTPVSEALTTMLNYIEVQDNGVRFIGA VL AGAALGVATGAAITAGI ALHK SNQNAQ Al SQMKD AIKNTNE AIQ SLKM ANQEMLS V VDSLRGQINTQIIPVMNQLGCQNIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPEVGYIALEIHFPTLTEIPNAVV QELMPISFNDKGDEWMTILPRYVLLRTTYTSNIDISKCLITDRSVICYNDYATPMSFDTIK CLTGNLTSCARERIIASHVPKFALSGGVIYANCLSVTCRCAIDGIPIVQSLRTTIMMLDNK KCRVYQIGELLISTGAYLGSIEFKNEDIELGPPIVLDpIDLGGQIAGINQTLQGVEDYIDKS NEILDKINPNVTSIGAMILVYIFIALSIVLGLSALIIGIKLNSQIKLLINQSIINQMRSGFDNA GYSRSLPSFSSTKSVNSTQELINTN

[0238] SEQ ID NO : 129, LechV- 1 (OR713879)

[0239] MEFKISFIIVSTLVIIATASLDYEELSKIGIIRGSTYNYKIKGSPSTKLMVVKLI PNLDRVENCTKTQLVNYKQLVKKALEPVKLSIDSMLSNVKSGNNKYRFAGAIMAGVAL GVATAATVTAGIALHQSNENARAIANLKNSIQSTNEAVKQLQMAGQQTLAVIDTIRGEI NNNIIPVINQLSCETVGLSVGIKLTQYYSEIITAFGPALQNPVNSRITIQAISSAFNGNFDEL LKTMGYSSNDMYEILQSGLIRGNIIDVDPDVGYIALEIEFPNLAPVPNAIIQELMPISFNVD GDEWVTLVPNFILTRTTLLSNIDVNRCTQTETSIICDHDYALPMSYQLMDCLKGSTDKCA REKVIS S YVPKF AL SGGVI YANCLNTICRCMDTDTPITQ SIK ST VTLLDNKNCEVYQ VGDI LISVGAYIGLHDYNSENVETGAPVVIDpIDIGNQLAGINQSLQNAEDYIEKSEEYLSGINPA IISIGSMVVLYIFLIIVAIIALIALVLSVKTTVKNSVIREQYAYRQQPPSIDNVNYVSR

[0240] SEQ ID NO: 130, LechV-2 (OR713880)

[0241] MEIKISFIIVSILVILATASLDYEELSKIGIIRGPTYNYKIKGSPSTKLMVVKLI PNLDRVENCTKTQLVNYKQLVKKALEPVKLSIDSMLSNVKSGNNKYRFAGAIMAGVAL GVATAATVTAGIALHQSNENARAIANLKNSIQSTNEAVKQLQMAGQQTLAVIDTIRGEI NNNIIPVINQLSCETVGLSVGIKLTQYYSEIITAFGPALQNPVNSRITIQAISSAFNGNFDEL LKTMGYSSNDMYEILQSGLIRGNIIDVDPDVGYIALEIEFPNLAPVPNAIIQELMPISFNVD GDEWVTLVPNFILTRTTLLSNIDVNRCTQTETSIICDHDYALPMSYQLMDCLKGSTDKCA REKVIS S YVPKF AL SGGVIYANCLNTICRCMDTDTPITQ SIK ST VTLLDNKNCEVYQ VGDI LISVGAYIGLHDYNSENVETGAPVVIDpIDIGNQLAGINQSLQNAEDYIERSEEYLSGINPA IISIGSMVVLYIFLIIVAIIALIALVLSVKTTVKNSVIREQYAYRQQPPSIDNVNYVSR

[0242] SEQ ID NO: 131, NinV (OQ438286)

[0243] MSENSHWYNIIYIIMKSRCSRLIGLIIVLTALMSAVVGIDFEGLSSIGVVKGR TLNYKIRGNSAYKILVIKLTPTIRSTSGAVNFENCTKDQISAHKVLVRNVLTPVKDALTS MKSKVTDYTPNSRLFGAIVAGAALVTATAATITAGVALHQSNQNAQAIANMKDAISKT NQAVSELKQGTQKLATVVDSLQNQINSQIIPVLNQLGCQTTGLTIGLYLTRYYSEILTVFG PAIQDPVNSALTIQAISKAFEGNFDKLMSAMGYSVSDLKDVLESDLVRGKIIDVDPDVGY MALQIEFPQITPVTGAHIQELLPISFNYKGDEWITILPSFVLNRMTYLSNIDIAHCLVTETS VICDNDLAMPMSTQMRDCLLGNTSKCSREQVITSYVPRFALSGGVIYANCLNTQCACAT NGRSISQSSRQTIMMLDDKDCKIYEVGGIFISVSTYMGIGKFENENISIGPPVVVDpIDVSGQLAEVNQSLSKTEELLEESNEYLSQVHVSLVSLKSMIILYVFIGLIGLLAAAALLVSVRSL SMLKTMGPATLMNNIQPTMHNLQYSGHSTINSKSSLINTHQSSHNHKYGSVNGSYTSDE S

[0244] Additional ectodomains from F proteins from additional viruses are shown below (SEQ ID NOs: 132-193). These proteins have a proline at the position equivalent to position 447 in AngV.

[0245] SEQ ID NO: 132, NiV-Ml (NC 002728)

[0246] MVVILDKRCYCNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVY VLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDPVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSL

[0247] SEQ ID NO: 133, NiV-M2 (AF376747)

[0248] MVVILDKRCYCNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPTTNNMRECLT GSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDPVDISSQISSMNQSLQQSKDYIKEAQR LLDTVNPSL

[0249] SEQ ID NO: 134, NiV-M3 (AJ627196)

[0250] MVVILDKRC YCNLLILILMISEC S VGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVY VLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYAIEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDPVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSL

[0251] SEQ ID NO: 135, NiV-M4 (FN869553)

[0252] MVVILDKRCYSNLLILILMISECSVGILHYEKLSKIGLVKGVTRKYKIKSNPLT KDIVIKMIANVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGV IMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVL TALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQ AFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQEL LPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTG STEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPT AVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQKSSMNQSLQQSKDYIKEAQRL LDTVNPSL

[0253] SEQ ID NO: 136, NiV-C (MK801755)

[0254] MVVVLDKRCYSNLLMLILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNP LTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVIKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPTAVLGNVIISLGKYLGSVNYNSEGIAVGPPVFTDpVDISSQISSMNQSLQQSKDYIKEA QRLLDTVNPSL

[0255] SEQ ID NO: 137, NiV-Bl.l (AY988601)

[0256] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYSSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEGFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSL

[0257] SEQ ID NO: 138, NiV-B1.2 (MK673568)

[0258] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPL TKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSSYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPT A VLGN VIISLGKYLGS VNYNSEGIAIGPP VFTDp VDI S SQI S SMNQ SLQQ SKD YIKEAQ RLLDTVNPSL

[0259] SEQ ID NO: 139, NiV-B1.3 (JN808864)

[0260] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSL

[0261] SEQ ID NO: 140, NiV-B1.4 (MK673584)

[0262] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYV LTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQ ELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECL TGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSINYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSL

[0263] SEQ ID NO: 141, NiV-B2.3 (MK673579)

[0264] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKVGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLA GVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVY VLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAI SQAFGGNYETLLRTLGYTTEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYI QELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMREC LTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTT CPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQ RLLDTVNPSL

[0265] SEQ ID NO: 142, NiV-B3.1 (MW535746)

[0266] MAVILNMRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPL TKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILMPIKGALEIYKNNTHDLVGDVRLAG VIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQMELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSL

[0267] SEQ ID NO: 143, NiV-B2.8 (PV132707)

[0268] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPTAVLGNVIVSLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSL

[0269] SEQ ID NO: 144, NiV-B1.9 (PP981667)

[0270] MAVILNKGYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPTAVLGNVIISLGKYLGSVNYNSESIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSL

[0271] SEQ ID NO: 145, NiV-B2.13 (PP981682)

[0272] MAVILNKRYYSNLLILILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDVRLAGVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYPSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSL

[0273] SEQ ID NO: 146, NiV-I2.2 (OR820508)

[0274] MAVILNKRYYSNLLLLILMISECSVGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNMSQCTGSVMENYKTRLNGILTPIKGALEIYKNNTHDLVGDLRLAGVIMAGVAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDKISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIIYVDLSGYYIIVRVYFPILTEIQQAYIQELLPVSFNNDNSEWISIVPNFILVRNTLISNIEIGFCLITKRSVICNQDYATPMTNNMRECLTGSTEKCPRELVVSSHVPRFALSNGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCPTAVLGNVIISLGKYLGSVNYNSEGIAIGPPVFTDpVDISSQISSMNQSLQQSKDYIKEAQRLLDTVNPSL

[0275] SEQ ID NO: 147, HeV-al.l (NC_001906)

[0276] MATQEVRLKCLLCGIIVLVLSLEGLGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNVSKCTGTVMENYKSRLTGILSPIKGAIELYNNNTHDLVGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSIAGQIVYVDLSSYYIIVRVYFPILTEIQQAYVQELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLITKKSVICNQDYATPMTASVRECLTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCTTVVLGNIIISLGKYLGSINYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEA QKILDTVNPSL

[0277] SEQ ID NO: 148, HeV-al.2 (HM044318)

[0278] MATQEVRLKCLLCGIIVLVLSLEGLGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNVSKCTGTVMENYKSRLTGILSPIKGAIELYNNNTHDLVGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYVQELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLITKKSVICNQDYATPMTASVRECLTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCTTVVLGNIIISLGKYLGSINYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEA QKILDTVNPSL

[0279] SEQ ID NO: 149, HeV-a7 (PQ660692)

[0280] MATQEVRLKCLLCGIIVLVLSLEGLGILHYEKLSKIGLVKGITRKYKIKSNPLTKDIVIKMIPNVSNVSKCTGTVMENYKSRLTGILSPIKGAIELYNNNTHDLVGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDQISCKQTELSLDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQAFGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYVQELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLITKKSVICNQDYATPMTASVRECLTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCTTVVLGNIIISLGKYLGSINYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEA QKILETVNPSL

[0281] SEQ ID NO: 15O, HeV-|31 (MZ318101)

[0282] MATQRVKVSYLICGIVISALSLEGLGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNVSKCTGTVMENYKNRLTGILSPIKGAIELYNNNTHDLIGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAISQA FGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYVQELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLTTKKSVICNQDYATPMTSTVRECLT GSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTC TTVVLGNIIISLGKYLGSTNYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKEAQR ILDTVNPSL

[0283] SEQ ID NO : 151 , HeV-p2 (MZ229747)

[0284] MATQGVKVSYLICGIVISALSLEGLGILHYEKLSKIGLVKGVTRKYKIKSNPLTKDIVIKMIPNVSNVSKCTGTVMENYKNRLTGILSPIRGAIELYNNNTHDLIGDVKLAGVVMAGIAIGIATAAQITAGVALYEAMKNADNINKLKSSIESTNEAVVKLQETAEKTVYVLTALQDYINTNLVPTIDQISCKQTELALDLALSKYLSDLLFVFGPNLQDPVSNSMTIQAIS QAFGGNYETLLRTLGYATEDFDDLLESDSITGQIVYVDLSSYYIIVRVYFPILTEIQQAYV QELLPVSFNNDNSEWISIVPNFVLIRNTLISNIEVKYCLTTKKSVICNQDYATPMTSTVRE CLTGSTDKCPRELVVSSHVPRFALSGGVLFANCISVTCQCQTTGRAISQSGEQTLLMIDNTTCTTVVLGNIIISLGKYLGSTNYNSESIAVGPPVYTDpVDISSQISSMNQSLQQSKDYIKE AQRILDTVNPSL

[0285] SEQ ID NO : 152, CedV- 1 (JQOO 1776)

[0286] MSNKRTTVLIIISYTLFYLNNAAIVGFDFDKLNKIGVVQGRVLNYKIKGDPMTKDLVLKFIPNIVNITECVREPLSRYNETVRRLLLPIHNMLGLYLNNTNAKMTGLMIAGVIMGGIAIGIATAAQITAGFALYEAKKNTENIQKLTDSIMKTQDSIDKLTDSVGTSILILNK LQTYINNQLVPNLELLSCRQNKIEFDLMLTKYLVDLMTVIGPNINNPVNKDMTIQSLSLL FDGNYDIMMSELGYTPQDFLDLIESKSITGQIIYVDMENLYVVIRTYLPTLIEVPDAQIYEF NKITMSSNGGEYLSTIPNFILIRGNYMSNIDVATCYMTKASVICNQDYSLPMSQNLRSCYQGETEYCPVEAVIASHSPRFALTNGVIFANCINTICRCQDNGKTITQNINQFVSMIDNSTC NDVMVDKFTIKVGKYMGRKDINNINIQIGPQIIIDpVDLSNEINKMNQSLKDSIFYLREAK RILDSVNISL

[0287] SEQ ID NO: 153, GhV(+A) (HQ660129)

[0288] MVINGNIITIILLLILILKTQMSEGAIHYETLSKIGLIKGITREYKVKGTPSSKDIVIKLIPNVTGLNKCTNISMENYKEQLDKILIPINNIIELYANSTKSAPGNARFAGVIIAGVA LGVAAAAQITAGIALHEARQNAERINLLKDSISATNNAVAELQEATGGIVNVITGMQDYINTNLVPQIDKLQCSQIKTALDISLSQYYSEILTVFGPNLQNPVTTSMSIQAISQSFGGNIDL LLNLLGYTANDLLDLLESKSITGQITYINLEHYFMVIRVYYPIMTTISNAYVQELIKISFNV DGSEWVSLVPSYILIRNSYLSNIDISECLITKNSVICRHDFAMPMSYTLKECLTGDTEKCP REAVVTSYVPRFAISGGVIYANCLSTTCQCYQTGKVIAQDGSQTLMMIDNQTCSIVRIEEI LISTGKYLGSQEYNTMHVSVGNPVFTDpLDITSQISNINQSIEQSKFYLDKSKAILDKINLN L

[0289] SEQ ID NO: 154, AngV (ON613535)

[0290] MGFRKNLRLLGIFWLIILSDTVDFERLASIGGIKGHSKLYKIKGHPTTKDIVIKLVPNLNNLTVCSEMSIDGYLNLINAVIIPISQSLELMRNNVKDGTPNENIFGAIVAGAAL GIATAAQVTSVVALHKSNQNAAKINSLRDSITKTNQAVEQLSLGVQETVSVLMGLQDQI NTNLVPKINILSCKQLSNTLNIMLLQYYSQILTVFGPNLRDPATVPVSIQALSQLFEGNLE LLTSSLGISSTDFNDLLTGKLITGSIIWADTQGHYLILRVNIPDLIEVPGAVIQEFIKIGYNF GGSLWMPTIPNKIIIRGYHLSLIDSTNCIITDNSYVCDRDYSLPMNPILRECFEGNTSSCGR EMVLNSYIPLYALSEGVIYANCLATSCKCATTNKPIVQSTSTVITMIDNSKCPVVEVGKQ MISVGSYLGQVSPNNLTIEIGPPVYTEPVDITNQLGNINETLSKTLDLIKQSNSILDMITGGL

[0291] SEQ ID NO: 155, LayV-Cl (OM101125)

[0292] MAFLKSAIICYLLFYPHIVKSSLHYDSLSKVGIIKGLTYNYKIKGSPSTKLM VVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNAMLDNVKSGNNKYRFAGAI MAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQNTNEAVKQLQLANKQTLAV IDTIRGEINNNIIPVINQLSCDTIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISSVFN RNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQELM PISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIGCLQ GDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTVSLLDNKKCLVYQVGDILISVGSYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIEKS EEFLKGINPSI

[0293] SEQ ID NO : 156, Lay V-C2 (OM 101130)

[0294] MAFLKSAIICYLLFYPHIVKSSLHYDSLSKVGIIKGLTYNYKIKGSPSTKLMVVKLIPNIDRVRNCTQKQFDEYKNLVKNVLEPVKLALNAMLDNVKSGNNKYRFAGAIM AGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQNTNEAVKQLQLANKQTLAVID TIRGEINNNIIPVINQLSCDTIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISSVFNR NFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQELMPI SYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIGCLQGDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTVSLLDNKKCL VYQVGDILISVGSYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIEKSEE FLKGINPSI

[0295] SEQ ID NO: 157, LayV-K2 (PQ641429)

[0296] MGRVVSLKLTLIWYLLLYSCIVETSLHYDSLSKVGIIKGLTYNYKIKGSPSTKLMVVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNTMLDNVKSGNNKYRFA GAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQSTNEAVKQLQLANKQTL AVIDTIRGEINNNIIPVINQLSCETIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISS VFNRNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQ ELMPISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIGCLHGDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTISLLDN KRCLVYQVGDILISVGTYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIE KSEEFLKGINPSI

[0297] SEQ ID NO: 158, LayV-K1.2 (PQ641427)

[0298] MERVVFLKLTLIWYLLLYSCTVETSLHYDSLSKVGIIKGLTYNYKIKGSPSTKLMVVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNTMLDNVKSGNNKYRFA GAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQSTNEAVKQLQLANKQTL AVIDTIRGEINNNIIPVINQLSCETIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISS VFNRNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQ ELMPISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIGCLQGDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTISLLDNKKCLVYQVGDILISVGTYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIE KSEEFLKGINPSI

[0299] SEQ ID NO: 159, LayV-K1.3 (PQ641428)

[0300] MGRVVYLKLTLIWYLLLYSCIVETSLHYDSLSKVGIIKGLTYNYKIKGSPST KLMVVKLIPNIDGVRNCTQKQFDEYKNLVKNVLEPVKLALNTMLDNVKSGNNKYRFA GAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKNAIQSTNEAVKQLQLANKQTL AVIDTIRGEINNNIIPVINQLSCETIGLSVGIKLTQYYSEILTAFGPALQNPVNTRITIQAISS VFNRNFDELLKIMGYTSGDLYEILHSGLIRGNIIDVDVEAGYIALEIEFPNLTLVPNAVVQ ELMPISYNVDGDEWVTLVPRFVLTRTTLLSNIDTSRCTVTESSVICDNDYALPMSYELIG CLQGDTSKCAREKVVSSYVPRFALSDGLVYANCLNTICRCMDTDTPISQSLGTTISLLDN KRCLVYQVGDILISVGTYLGEGEYSADNVELGPPVVIDpIDIGNQLAGINQTLQNAEDYIE KSEEFLKGINPSI

[0301] SEQ ID NO: 160, Moj V (NC_025352)

[0302] MALNKNMF S SLFLGYLLVYATT VQ S SIHYD SL SK VGVIKGLT YNYKIKGSP STKLMVVKLIPNIDSVKNCTQKQYDEYKNLVRKALEPVKMAIDTMLNNVKSGNNKYRF AGAIMAGVALGVATAATVTAGIALHRSNENAQAIANMKSAIQNTNEAVKQLQLANKQ TLAVIDTIRGEINNNIIPVINQLSCDTIGLSVGIRLTQYYSEIITAFGPALQNPVNTRITIQAIS SVFNGNFDELLKIMGYTSGDLYEILHSELIRGNIIDVDVDAGYIALEIEFPNLTLVPNAVV QELMPISYNIDGDEWVTLVPRFVLTRTTLLSNIDTSRCTITDSSVICDNDYALPMSHELIG CLQGDTSKCAREKVVSSYVPKFALSDGLVYANCLNTICRCMDTDTPISQSLGATVSLLD NKRCSVYQVGDVLISVGSYLGDGEYNADNVELGPPIVIDpIDIGNQLAGINQTLQEAEDY IEKSEEFLKGVNPSI

[0303] SEQ ID NO : 161 , SHNV5 - 1 (OQ715593 )

[0304] MINMDSKRLKVKVVFPIYLIIYIQMSRASLDYDQLSKIGIVKGPSYNYKIKG SPSTKLMVVKLVPNIKEVANCTHKQVENYKTLVRNVLDPVKMSLQAMLEKVKSGNNK YRFAGAIMAGVALGVATAATVTAGIALHRSSENAQAIARIKGAIQNTNEAVKQLQLAN KQMLAVIDTIRGEINENIIPIMNELSCETIGLNVGIKLTQYYSEIITAFGPALQNPIGSKITIQ AISSAFNGNFDELLKSMGYTSNDLYEVLQSGLIRGNIIDVDPEVGYIALEIEFPNLTLVPNAIIQELMPISFNSEGDEWVTLMPRFILTRTTLLSNIDITKCTVTDRSVICDNDYALPMSNQL IECLRGDTMKCTREKVMSSYIPKFALSDGVVYANCLNTICRCMDTDTPITQSLRSTVTLL DNKACLVYQIGDILISVGSYLGNTEYNTQNITLGPPIVIDpIDIGNQLAGINQSLQNAEDYI EKSDEFLKGINPSV

[0305] SEQ ID NO: 162, SHNV5-2 (MZ328275)

[0306] MINMESKRLKVKVVFSIYFIIYIQMSRASLDYDQLSKIGIVKGPSYNYKIKG SPSTKLMVVKLVPNIKEVANCTHKQIENYKTLVRNVLDPVKMSLQAMLEKVKSGNNKY RFAGAIMAGVALGVATAATVTAGIALHRSSENAQAIARIKGAIQNTNEAVKQLQLGNK QMLAVIDTIRGEINENIIPIMNELSCETIGLNVGIKLTQYYSEIITAFGPALQNPIGSKITIQAI SSAFNGNFDELLKSMGYTSNDLYEVLQSGLIRGNIIDVDPEAGYIALEIEFPNLTLVPNAII QELMPISFNSEGDEWVTLMPRFILTRTTLLSNIDITKCTVTDRSVICDNDYALPMSNQLIE CLRGDTMKCTREKVMSSYIPKFALSDGVVYANCLNTICRCMDTDTPITQSLRSTVTLLDNKACLVYQIGDILISVGSYLGNTEYNTQNITLGPPIVIDpIDIGNQLAGINQSLQNAEDYIE KSEEFLKGINPSV

[0307] SEQ ID NO : 163 , Has V (OR713881)

[0308] MTIVNTVNMKLVLLVIYLIISSEYVKASLDYNELSKVGVIKGLSYNYKIRGSPSKKLMVVKLIPNIDTVENCSKTQLENYKILVRNALEPVKLSIKAMLDNVKSGNNKYRF AGAIMAGVALGVATAATVTAGIALHQSKENAQAIANIKSAIQNTNEAVKQLQLANKQT LAVIDTIRGEINDNIIPVINQLSCQTIGLNIGIKLTQYYSEILTAFGPAMQNPVNSRITIQAIS STFNGNFDELIKIMGYTSSDLYEVLHSGLIRGNIIDVDPEVGYIALEIEFPNLTLVPNAIIQE LMPISFNIDGDEWVSLVPRFVLTRTTLLSNIDTDKCTVTEKSVICDNDYALPMSYQLVEC LTGDTSKCTREKVISTYVPRFALSDGLIYANCLNTICRCMDTDTPIAQGLKSTVSLLDNK NCLVYQVGDILISVGTYLGETEYNTENIQLGPPVVIDpIDIGSQLAEINKTLQSAEDYIEKSDEFLKGVNPSV

[0309] SEQ ID NO: 164, MelV (OK623353)

[0310] MAKFVFLKTMCIGLLIIISDRVDSSMDFHSLSKIGIIKGKTYNYKIRGEPNTK LMVIKLIPNIDVVENCSTTQVNNYKKLVRNVLTPVRIISRYYVKNVIEQNNRVRLFGAIM AGAALGVATAATVTAGIALHRSNENARNIALMKEAIKNTNQAVTKLQLAGQQTLAVIDNIRGEINNQIIPVINKLTCENIGLNVGIKLTQYYSEVLTAFGPAIQDPVNARITIQAISKVFN NNFDELLNVMGYSTQDLYEVLHGGLIRGNVISADPEVGYLALEIEFPNLSVVPNAYIQEI MPISFNVDGDEWVTVVPRHTLIRTTLLSNIDITTCSIIESSIICNNDYALPMSNELINCLQGS TDLCAREKVISNYVPKFALSDGVVYANCLSTVCRCMDNGVPISQSLKSTVMMLDDKKC TIYQIGDILISVGKYMGHIDYNPENVVLGPPIVIDpIDIGNQLAGINQTLQEAGDFIEKSEEI LNSINPSV

[0311] SEQ ID NO: 165, DewV-1 (OK623354)

[0312] MARLQVVILYLYLLVASDVVKASLDFDNMSKIGIIKGNTYNYKIRGEPTTK LMVVKLIPNIDVVENCSATQLANYKKLVTNVLTPVKLSLDNMLKNVQDQNNRVRLFGA IMAGAALGVATAATVTAGIALHRSNENAKNIAKLKNAIQNTNLAITKLQMAGQQTLAVI DNIRGEINNQIIPVLNQLSCETVGLNVGIKLTQYYSEVLTAFGPAIQDPVNSKITIQAISKA FNNNFDELLKVMGYTSQDLYEILHGDLITGNVISADPEAGYIALEIEFPNLSTVPNAYVQE LMPISFNVDGDEWVTLVPRYVLIRTTFLSNIDISLCSIMETSIVCNNDYALPMSSELINCLQ GDTGVCAREKVISSYVPKFALSGGVVYANCLSTVCRCMDNGTPISQGIRHTVALLDNKK CSVYQVGDILISVGKYLGHLDYNTEDIVLGPPIVIDpIDIGNQLAGINSSLQQAENYIDKSN EILKSINPTI

[0313] SEQ ID NO: 166, DewV-2.1 (OR713882)

[0314] M A K LQM T 1 F YL YLL VASDL VET SLDFDNMSKIGIIKGNTYNYKIRGEPTTKLMVVKLIPNIDVVENCSATQLANYKKLVNNVLTPVKLSLDNMLKNVQDQNNRVRLFG AIMAGAALGVATAATVTAGIALHRSNENAKNIAKLKNAIQNTNLAITKLQMAGQQTLA VIDNIRGEINNQIIPVLNQLSCETVGLNVGIKLTQYYSEVLTAFGPAIQDPVNSKITIQAISK AFNNNFDELLKVMGYTSQDLYEILHGDLITGNVISADPETGYIALEIEFPNLSTVPNAYVQ ELMPISFNVDGDEWVTLVPRYVLIRTTFLSNIDISLCSIMETSIVCNNDYALPMSSELINCL QGDTGVCAREKVISSYVPKFALSGGVVYANCLSTVCRCMDNGTPISQGIKHTVALLDNK KCTVYQVGDILISVGKYLGHLDYNTEDIVLGPPIVIDpIDIGNQLAGINSSLQQAEDYIDKS NE1LKGINPTI

[0315] SEQ ID NO: 167, DewV-2.2 (OR713883)

[0316] MAKLQMTIFYLYLLVASDLVETSLDFDNMSKIGIIKGNTYNYKIRGEPTTK LMVVKLIPNIDVVENCSATQLANYKKLVNNVLTPVKLSLDNMLKNVQDQNNRVRLFG AIMAGAALGVATAATVTAGIALHRSNENAKNIAKLKNAIQNTNLAITKLQMAGQQTLAVIDNIRGEINNQIIPVLNQLSCETVGLNVGIKLTQYYSEVLTAFGPAIQDPVNSKITIQAISK AFNNNFDELLKVMGYTSQDLYEILHGDLITGNVISADPEAGYIALEIEFPNLSTVPNAYV QELMPISFNVDGDEWVTL VPRYVLIRTTFL SNIDISLC SVMET SIVCNNDYALPMS SELIN CLQGDTGVCAREKVISSYVPKFALSGGVVYANCLSTVCRCMDNGTPISQGIKHTVALLD NKKCTVYQVGDILISVGKYLGHLDYNTEDIVLGPPIVIDpIDIGNQLAGINSSLQQAEDYI DKSNEILKGINPTI

[0317] SEQ ID NO: 168, DarV-K (MZ574409)

[0318] MSSSNKIRITVIIINILISNYLIHCSMDFVELSRVGIIKGSTYNYKIRGEPSTKLMVVKLIPNVGDVANCSATQVSNYKKLVKNVLSPVSNALNTMLENVQTENNRYRLFGAI MAGAALGVATAATVTAGIALHRSNENARNIAAMKNAIQNTNQAITKLQMAGQQTLAVI DNIRGEINNQIIPLLNKLSCETVGLNVGIKLTQYYSEILTVFGPAIQDPINSKITIQAISKAFG GNFDELLKVMGYTSQDLYEILHGGLITGNIIGVDPDTGYIALEIEFPNLSIVQNAYIQELM PISFNVDGDEWVTLAPRFVLIRTTLLSNIDTSMCTIIDSSVICNNDYALPMSTELINCLQG MTESCAREKVISSYVPKFALSGGVIYANCLSTVCRCMDNQKPISQSLRSTVVMLDNKMCKVYQIGDILISVGEYKGAVEYNPEDVHLGPPIVLDpIDIGNQLAGINQTLSEAGDFISKSEE ILKDINPAI

[0319] SEQ ID NO: 169, DarV-C (OM030315)

[0320] MSSSNKIRITVIIINILISNYLINCSMDFVELSRVGIIKGSTYNYKIRGEPSTKLMVVKLVPNVGDVANCSTTQVSNYKKLVKNVLSPVSNALNTMLENVQTENNRYRLFGA IMAGAALGVATAATVTAGIALHRSNENARNIAAMKNAIQNTNQAITKLQMAGQQTLAV IDNIRGEINNQIIPLLNKLSCETVGLNVGIKLTQYYSEILTVFGPAIQDPINSKITIQAISKAF GGNFDELLNVMGYTSQDLYEILHGGLITGNIIGVDPDTGYIALEIEFPNLSIVQNAYIQEL MPISFNVDGDEWVTLAPRFVLTRTTLLSNIDTSMCTIIDSSVICNNDYALPMSTELINCLQ GMTESCAREKVISSYVPKFALSGGVIYANCLSTVCRCMDNQKPISQSLRSTVVMLDNKMCKVYQIGDILISVGEYKGAVEYNPEDVHLGPPIVLDpIDIGNQLAGINQTLSEAGDFISKSE EILKDINPAI

[0321] SEQ ID NO: 170, DarV-Ca (PP272750)

[0322] MSSSNKIRITVIIINILISNYLINCSMDFVELSRVGIIKGSTYNYKIRGEPSTKLMVVKLIPNVGDVANCSATQVSNYKKLVKNVLSPVSNALNTMLENVQTENNRYRLFGAI MAGAALGVATAATVTAGIALHRSNENARNIAAMKNAIQNTNQAITKLQMAGQQTLAVI DNIRGEINNQIIPLLNKLSCETVGLNVGIKLTQYYSEILTVFGPAIQDPINSKITIQAISKAFG GNFDELLKVMGYTSQDLYEILHGGLITGNIIGVDPDTGYIALEIEFPNLSIVQNAYIQELM PISFNVDGDEWVTLAPRFVLTRTTLLSNIDTSMCTIIDSSVICNNDYALPMSTELINCLQG MTESCAREKVISSYVPKFALSGGVIYANCLSTVCRCMDNQKPISQSLRSTVVMLDNKMCKVYQIGDILISVGEYKGAVEYNPEDVHLGPPIVLDpIDIGNQLAGINQTLSEAGDFISKSEE ILKDINPAI

[0323] SEQ ID NO: 171, GakV-1 (MZ574407)

[0324] MELIKLFNILYLVGYSGLFFTDVNAGLDYEGLSSIGVIKGPSYNYKIRGTPS TKLLVIKLIPNVESIDNCTQKQMSDYRALVKNVLTPVSESLSTMLNYIEQQSNGVRLIGAVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKMANQELLGV VDSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYSGSDLHDILQGDLITGNIIGVDPDVGYIALEINFPTLTEIPNAVIQ ELMPISFNDKGDEWMALVPRYVLLRTTYISNIDISKCLLTERSVICYNDYATPMSFDIIRC LTGNLTYCPREQIMASHVPKF AL SGGVIYANCL S AVCRC AVDGVPIVQ SLK ATVMMLD NKSCRVYQIGELLISTGAYLGSIEFKNENIELGPPIVIDpIDLGGQIAGINQTLQGVEDYIDK SNEILDQVNPSV

[0325] SEQ ID NO: 172, GakV-2 (MZ574408)

[0326] MELIKLFYILYLVGYSGLFFTDVNAGLDYEGLSSIGVIKGPSYNYKIRGTPS TKLLVIKLIPNVESIDNCTQKQMSDYRALVKNVLTPVSESLSTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKMANQELLGV VDSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYSGSDLHDILQGDLITGNIIGVDPDVGYIALEINFPTLTEIPNAVIQELMPISFNDKGDEWMALVPRYVLLRTTYISNIDISKCLLTERSVICYNDYATPMSFDIIRC LTGNLT YCPREQIM ASHVPKF AL SGGVIYANCL S A VCRC AVDGVPI VQ SLK AT VMMLD NKNCRVYQIGELLISTGAYLGSIEFKNENIELGPPIVIDpIDLGGQIAGINQTLQGVEDYIDK SNDILDQVNPSV

[0327] SEQ ID NO : 173 , SHNV 1 (OQ236120)

[0328] MNYTIVGIMIITFVHESQCINYEQLASIGVIKGHTYNYKIRGPPNTKLMVVK LIPNINIDNLGGGLSNCSSKQMESHKELVEKVLSPVAQALETMRNRVTDYSGNYRFVGA VMAGAALGVATAATVTAGIALHQSNQNAKAIDQMKEAIRTTNKAVQELTLSTRQTLLV IDSLQNQINTQIVPAMNRLSCEVLGLTVGIQLTQYYSEILTYFGPALQDPIDSTLTIQAISH AFGGNFDILMKTMGYTVGDLYDVLKGDLITGKIISVNPKEGFIALEVRFPTLTQVNNAIV QELMPISFNDKGDEWISTVPRYVLERVLYLSNIDISLCSVGETSVVCDNDYASPMSHQLR ECLQTNTSYCPRERVLASYVPKFALSQGVIFANCIATTCRCADDGRAISQSSSQTVLLLTS KDCKVYEVQSMMISTGEYLGESIFENTDIPLGPSIVIDpIDISGQLAEINKTLDHVDNTIKDSNDILDKIDVST

[0329] SEQ ID NO : 174, SHNV2 (OM030316)

[0330] MSLKFRQSTNNRTNIRLILIIVLIKHMLCIHYENLSSVGIIKGNTYNYKIKGDPSTKMLVVKLIPNIDGLGNCTDKQMKDYKTLVKSTLQPVKDSLAQMLNNVETYNGYV KFFGAVMAGAALGVATAATVTAGIALHQSNQNAKDIANMKDAIMKTNQAITTLSSASQ KMLTVIDSIRGEINQQIIPVLNQLSCETVGLNLGIKLTQYYSQISTFFGPALQNPVQSILTIQ AISHAFGGNFNELMTVMGYTGSDFQDIIQGNLITGSVIGVDPEVGYIALEIRIPSLTVVPN AYVQELLPVSFNIDGDEWMTIVPNYVLTRTTYLSNIDINRCLITDKSVICGNDYATPMSN QLINCLNGNTQHCAREAVVTSYVPKFALSGGVIYANCLSTVCRCIDKDQPISQSLSQTLM MLDNQHCNVYQISNVLISTGRYLGDAEFRNEGIDLGPPIVVDpIDLGGQIADINQTISDAE EFIEESNKILSKINPKI

[0331] SEQ ID NO: 175, SHNV3 (OM030317)

[0332] MELIKNYCLIYLIFYSSTLIRPLQAGLDYEALASIGVIKGPSYNYKIRGTPST KLLVIKLIPNVGSLDNCTQKQMSDYRALVKNVLTPVSESLKTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDSIKNTNMAVQTLKLANQEILGVVDSLRGQINTQIIPVLNQLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDVGYIALEIHFPTLTEIPNAVIQ ELMPISFNDKGDEWMALVPRYVLLRTTYVSNIDISKCLLTERSVICYNDYATPMSFDVLR CLTGNLTFCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVDGVPIVQSLKTTIMMLDN KNCRVYQIGELLISTGAYLGSVEFKNENIELGPPIVVDpIDLGGQIAGINQTLQGVEDYID KSNEILDQVNPSV

[0333] SEQ ID NO : 176, SHNV4 (OM030314)

[0334] MGLTKSTILVYLLVYYHAHIIAINAGLDYEGLASIGVVKGPSYNYKIRGTPT TKLLVIKLIPNVGSLDNCTQKQMADYKSLVKNVLTPVSDALSTMLNYIEQQSNGVRLIG AVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKLANQEILG VVDSLRGQINTQIIPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQE LMPISFNDKGDEWMTLVPRYVLLRTTYVSNIDISKCLITERSVICYNDYATPMSFDVIRCL TGNLTYCPREQIIASYVPRFALSGGVIYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKN CRVYQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQGVEDYIDKSNEILDQVNPSV

[0335] SEQ ID NO: 177, SHNV6 (PQ541138)

[0336] MAKESTSICISYLLIYCHLISGNLDYEALSKIGIIKGPSYNYKIKGTPSTKLM VVKLIPNL SNVENC SRGQIENYRNL VKNVLDP VKNALD AMLENVKTGNNRYRF AGAIN! AGVALGVATAATVTAGIALHRSNENAQAIANMKSAIQSTNEAVKQLQTANQQMLAVID TVRGEINNNIIPVINQLSCETIGLNVGIKLTQYYSEVITAFGPALQNPVNTRITIQAISSAFN GNFDELLKIMGYTSNDLYEILQSGLIRGNIIDVDPIVGYIALEIEFPNLSLVPNAIIQELMPIS FNVDGDEWVSMVPYYVLTRTTLLSNIDINRCSVTTKSIICDNDYALPMSNQLIDCLKGET EKCARERVISSYVPKFALTDGVVYANCLHTICRCMDTDTPISQGLSSTVMLLDSKKCLV YQVGDILISVGEYLGESEYRTDNIELGPPIVIDpIDIGNQLAGINQTLQNAGDYIEKSEEFLSGVNPAV

[0337] SEQ ID NO: 178, SHNV7 (PQ541139)

[0338] MELVVIKKVIIIYILMYNHLIEASLDFNNLAKIGIIKGKTYNYKIRGEPNTKL MVVKLIPNIDVVQNCSGTQIANYKKLVNNVLSPIKLALDNMLKNVIEQNNRVRLFGAIM AGAALGVATAATVTAGIALHRSNENARNIALLKDSIKNTNQAVAKLQMANQQTLAVID NIRGEINNQIIPVMNKLTCETIGLNVGIKLTQYYSEILTVFGPAIQDPVNSKITIQAISKVFN NNFDELLNVMGYSSQDLYEILHGGLIRGNVISADPEIGYMALEIEFPNLAVVPNAYIQEIM PISFNVDGDEWVTVVPRYTLIRTTLLSNIDISLCTLIETSVICNNDYALPMSSELVNCLQGN TEVCAREKVISNYVPKFALSDGVVYANCLSTVCRCMDNGVPISQNLKSTVMMLDDKKC TIYQIGDILISVGKYMGHVEYNPEDVVLGPPIVIDpIDIGNQLAGINQTLQEAGDFIEKSEEILNSINPNV

[0339] SEQ ID NO: 179, SHNV8 (PP272530)

[0340] MKLTKYYCMLYLIGYAGFHFTEVDAGLDYEGLASIGVIKGPSYNYKIRGTPSTKLLVIKLIPNVDSIDNCTQKQMSDYKALVKNVLTPVSESLSTMLNYIEQQSNGVRLI GAVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIKNTNQAVQTLKMANQELL GVVDSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAI SGAFSGNFDEMMRVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEINFPTLTEIPNAII QELMPISFNDKGDEWMALVPRFVLLRTTYISNIDISKCLLTERSVICYNDYATPMSFDIIR CLTGNLTYCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVDGVPIVQSLKATVMMLD NKNCRVYQIGELLISTGTYLGSVEFRNENIELGPPIVVDpIDLGGQIAGINQTLQGVEDYID KSNEILDQVNPSV

[0341] SEQ ID NO: 180, SHNV9 (PP272531)

[0342] MKLIRCICLLYLIGYSGLLLVNVDAGLDYEGLASIGVIKGPSYNYKIRGTPSTKLLVIKLIPNVGSIDNCTQKQMSDYKALVKNVLTPVAESLSTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDAIRNTNQAVQTLKMANQELLGV VDSLRGQINTQIIPVLNKLSCDTVGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDVGYIALEINFPTLTEIPNAIIQE LMPISFNDKGDEWMALVPRYVLLRTTYISNIDTSKCLLTERSVICYNDYATPMSFDIIRCL TGNLTYCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVDGVPIVQSLKATVMMLDNKSCRVYQIGELLISTGTYLGSVEFRNENIELGPPIVVDpIDLGGQIAGINQTLQGVEDYIDKS NE1LEQVNPSV

[0343] SEQ ID NO: 181, SHNV10-1.2 (PP272749)

[0344] MELIK STIL VYLLIHQQLNIIT VS AGLDYEGLASIGVVKGPSYNYKIRGTPTTKLLVIKLIPNVGSLDNCTQKQMADYKALVKNVLTPVSDALSTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLRDAIKNTNQAVQTLKLANQELLGV VDSLRGQINTQIIPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAF SGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQEL MPISFNDKGDEWMTLVPRYVLLRTTYISNIDISKCLVTERSVICYNDYATPMSFDVIRCLT GNLTYCPREQIIASYVPRFALSGGVIYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKNC RVYQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQNVEDYIDKSNE ILDQVNPSV

[0345] SEQ ID NO: 182, SHNV10-2.1 (PP272534)

[0346] MELIKSTILIYLLIHQQLNIITVSAGLDYEGLASIGVVKGPSYNYKIRGTPTTKL LVIKLIPNVGSLDNCTQKQM AD YK AL VKNVLTP VSD AL STMLNYIEQQ SNGVRLIGAVL AGAALGVATGAAITAGIALHKSNQNAQAIAQLRDAIKNTNQAVQTLKLANQELLGVVD SLRGQINTQHPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAFSG NFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQELMPI SFNDKGDEWMTLVPRYVLLRTTYISNIDISKCLVTERSVICYNDYATPMSFDVIRCLTGN LTYCPREQIIASYVPRFALSGGVVYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKNCR VYQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQGVEDYIDKSNGI LDQVNPSV

[0347] SEQ ID NO: 183, SHNV10-2.2 (PP272638)

[0348] MELIKSTILIYLLIHQQLNIITVSAGLDYEGLASIGVVKGPSYNYKIRGTPTT KLLVIKLIPNVGSLDNCTQKQMADYKALVKNVLTPVSDALSTMLNYIEQQSNGVRLIGA VLAGAALGVATGAAITAGIALHKSNQNAQAIAQLRDAIKNTNQAVQTLKLANQELLGV VDSLRGQINTQIIPVINQLSCDTIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAF SGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDIGYIALEIHFPTLTEIPNAVIQELMPISFNDKGDEWMTLVPRYVLLRTTYISNIDISKCLVTERSVICYNDYATPMSFDVIRCLT GNLTYCPREQIIASYVPRFALSGGVVYANCLSTVCRCAVDGVPIVQSLKATIMMLDNKN CRVYQIGELLISTGAYLGSVEFKNENIDLGPPIVIDpVDLGGQIAGINQTLQGVEDYIDKSN EILDQVNPSV

[0349] SEQ ID NO : 184, SHNV 11 (OQ970176)

[0350] MKLNNTRRDNKINKQHTTMACIKNCCLVYLILYSSTLIIPLEAGLDYESLASIG VIKGPSYNYKIRGTPSTKLLVIKLIPNVGSLDNCTQKQMADYKALVKNVLTPVSESLKT MLNYIEQQSNGVRLIGAVLAGAALGVATGAAITAGIALHKSNQNAQAIAQLKDSIKNTN MAVQTLKLANQEILGVVDSLRGQINTQIIPVLNQLSCDTVGLTLGIKLTQYYSEILTAFGP AIQDPVNSKLTIQAISGAFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPDVGYI ALEIHFPTLTEIPNAVIQELMPISFNDKGDEWMALVPRYVLLRTTYVSNIDISKCLLTERS VICYNDYATPMSFDVLRCLTGNLTFCPREQIIASHVPKFALSGGVIYANCLSAVCRCAVD GVPIVQSLKTTIMMLDNKNCRVYQIGELLISTGSYLGSVEFKNENIELGPPIVVDpIDLGGQIAGINQTLQGVEDYIDKSNEILDQVNPSV

[0351] SEQ ID NO: 185, ChV-1 (PQ140950)

[0352] MARWLRNFIILYLIVFYSEYSGALHYENLSYVGVIKGLTYNYKIKGDPSTK LLVIKLIPNLNITKENFNDCTRKQMDEHTKLVRSVLEPVKAALDAMRNKVTDYSGNYRF FGAVLAGAAMGVATAATITAGIALHKANENAYAINQMKDAIKNTNKAIQEVSNAQRQT VVVLDNLQGQINTQIIPVLNQLTCEMQGLTVGLQLTRYYSEILTFFGPAIQDPVNSVLTIQ AISHAFDRNFDALMTTMGYTAADMYEVLNSDLITGRIISVDPTIGYIALEIRFPTLTTIENA IIQELMPISFNHKSDEWITIVPKYVLERMSFISNIDTSLCLVGDKSIICDNDYATPMSTQMR NCLEGNTESCVREKVLASYVPKFALSGGVIYANCISAACRCADNGEAISQSSSSSVMMLS NQGCSTYEVQNTLISVGKYMGEKEFNSMDIEVGPPIVLDpVDISGQLTEINKTLDKAEEYIEESNDILKGIQVSL

[0353] SEQ ID NO: 186, ChV-2.1 (PQ140949)

[0354] MARWSRNFIILYMILFYSEYSGALHYENLSYVGVIKGLTYNYKIKGDPSTK LLVIKLIPNLNITKENFNDCTRKQMDEHTKLVRSVLEPVKAALDAMRNKVTDYSGNYRF FGAVLAGAAMGVATAATITAGIALHKANENAYAINQMKDAIKNTNKAIQEVSNAQRQTVVVLDNLQGQINTQIIPVLNQLTCEMQGLTVGLQLTRYYSEILTFFGPAIQDPVNSVLTIQAISHAFDRNFDALMTTMGYTAADMYEVLNSDLITGRIISVDPTIGYIALEIRFPTLTTIENAIIQELMPISFNHKSDEWITIVPKYVLERMSFISNIDTSLCLVGDKSIICDNDYATPMSTQMRNCLEGNTESCVREKVLASYVPKFALSGGVIYANCISAACRCADNGEAISQSSSSSVMMLSNQGCSTYEVQNTLISVGKYMGEKEFNSMDIEVGPPIVLDpVDISGQLTEINKTLDKAEDY IEESNDILKGIQVSL

[0355] SEQ ID NO: 187, ChV-2.2 (PQ140951)

[0356] MARWLRNFIILYMILFYSEYSGALHYENLSYVGVIKGLTYNYKIKGDPSTKLLVIKLIPNLNITKENFNDCTQKQMDEHTKLVRSVLEPVKAALDAMRNKVTDYSGNYRFFGAVLAGAAMGVATAATITAGIALHKANENAYAINQMKDAIKNTNKAIQEVSNAQRQTVVVLDNLQGQINTQIIPVLNQLTCEMQGLTVGLQLTRYYSEILTFFGPAIQDPVNSVLTIQAISHAFDRNFDALMTTMGYTAADMYEVLNSDLITGRIISVDPTIGYIALEIRFPTLTTIENAIIQELMPISFNHKSDEWITIVPKYVLERMSFISNIDTSLCLVGDKSIICDNDYATPMSTQMRNCLEGNTESCVREKVLASYVPKFALSGGVIYANCISAACRCADNGEAISQSSSSSVMMLSNQGCSTYEVQNTLISVGKYMGEKEFNSMDIEVGPPIVLDpVDISGQLTEINKTLDKAEEYIEESNDILKGIQVSL

[0357] SEQ ID NO: 188, ResV-a (OR713876)

[0358] MKFLIIKYVTINYMLIYVSIFVKVSRSNLDYEALSSIGVIKGPSYNYKIRGNPITKLLVIKLIPNVGNISDCTTKQIQDYKTLVRNVLTPVSEALTTMLNYIEVQDNGVRFIGAVLAGAALGVATGAAITAGIALHKSNQNAQAISQMKDAIKNTNEAIQSLKMANQEMLSVVDSLRGQINTQIIPVMNQLGCQNIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISGAFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPEVGYIALEIHFPTLTEIPNAVVQELMPISFNDKGDEWMTILPRYVLLRTTYTSNIDISKCLITDRSVICYNDYATPMSFDTIKCLTGNLTSCARERIIASHVPKFALSGGVIYANCLSVTCRCAIDGIPIVQSLRTTIMMLDNKKCRVYQIGELLISTGAYLGSIEFKNEDIELGPPIVLDpIDLGGQIAGINQTLQGVEDYIDKSNDI LDKINPNV

[0359] SEQ ID NO: 189, ResV-01 (OR713877)

[0360] MRFVIIKYVTINYMLIYVSVFVKVSRSNLDYEALSSIGVIKRASYNYKIRGN PITKLLVIKLIPNVGNISDCTTKQIQDYKTLVRNVLTPVSEALTTMLNYIEVQDNGVRFIG AVLAGAALGVATGAAITAGIALHKSNQNAQAISQMKDAIKNTNEAIQSLKMANQEMLS VVDSLRGQINTQIIPVMNQLGCQNIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAIS GAFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPEVGYIALEIHFPTLTEIPNAV VQELMPISFNDKGDEWMTILPRYVLLRTTYTSNIDISKCLITDRSVICYNDYATPMSFDTI KCLTGNLTSCARERIIASHVPKFALSGGVIYANCLSVTCRCAIDGIPIVQSLRTTIMMLDN KKCRVYQIGELLISTGAYLGSIEFQNEDIELGPPIVLDpIDLGGQIAGINQTLQGVEDYIDKSNEILDKINPNV

[0361] SEQ ID NO: 190, ResV- 2 (OR713878)

[0362] MRFFITRYVIINYMLIYISFFVKVGRSNLDYEALSSIGVIKGASYNYKIRGNPITKLLVIKLIPNVGNISDCTTKQIQDYKTLVRNVLTPVSEALTTMLNYIEVQDNGVRFIGA VLAGAALGVATGAAITAGIALHKSNQNAQAISQMKDAIKNTNEAIQSLKMANQEMLSV VDSLRGQINTQIIPVMNQLGCQNIGLTLGIKLTQYYSEILTAFGPAIQDPVNSKLTIQAISG AFSGNFDEMMKVMGYTGSDLHDILQGDLITGNIIGVDPEVGYIALEIHFPTLTEIPNAVV QELMPISFNDKGDEWMTILPRYVLLRTTYTSNIDISKCLITDRSVICYNDYATPMSFDTIK CLTGNLTSCARERIIASHVPKFALSGGVIYANCLSVTCRCAIDGIPIVQSLRTTIMMLDNK KCRVYQIGELLISTGAYLGSIEFKNEDIELGPPIVLDpIDLGGQIAGINQTLQGVEDYIDKS NEILDKINPNV

[0363] SEQ ID NO : 191 , LechV- 1 (OR713879)

[0364] MEFKISFIIVSTLVIIATASLDYEELSKIGIIRGSTYNYKIKGSPSTKLMVVKLIPNLDRVENCTKTQLVNYKQLVKKALEPVKLSIDSMLSNVKSGNNKYRFAGAIMAGVAL GVATAATVTAGIALHQSNENARAIANLKNSIQSTNEAVKQLQMAGQQTLAVIDTIRGEI NNNIIPVINQLSCETVGLSVGIKLTQYYSEIITAFGPALQNPVNSRITIQAISSAFNGNFDEL LKTMGYSSNDMYEILQSGLIRGNIIDVDPDVGYIALEIEFPNLAPVPNAIIQELMPISFNVD GDEWVTLVPNFILTRTTLLSNIDVNRCTQTETSIICDHDYALPMSYQLMDCLKGSTDKCA REKVISSYVPKFALSGGVIYANCLNTICRCMDTDTPITQSIKSTVTLLDNKNCEVYQVGDILISVGAYIGLHDYNSENVETGAPVVIDpIDIGNQLAGINQSLQNAEDYIEKSEEYLSGINPA I

[0365] SEQ ID NO: 192, LechV-2 (OR713880)

[0366] MEIKISFIIVSILVILATASLDYEELSKIGIIRGPTYNYKIKGSPSTKLMVVKLI PNLDRVENCTKTQLVNYKQLVKKALEPVKLSIDSMLSNVKSGNNKYRFAGAIMAGVAL GVATAATVTAGIALHQSNENARAIANLKNSIQSTNEAVKQLQMAGQQTLAVIDTIRGEI NNNIIPVINQLSCETVGLSVGIKLTQYYSEIITAFGPALQNPVNSRITIQAISSAFNGNFDEL LKTMGYSSNDMYEILQSGLIRGNIIDVDPDVGYIALEIEFPNLAPVPNAIIQELMPISFNVD GDEWVTLVPNFILTRTTLLSNIDVNRCTQTETSIICDHDYALPMSYQLMDCLKGSTDKCA REKVISSYVPKFALSGGVIYANCLNTICRCMDTDTPITQSIKSTVTLLDNKNCEVYQVGDI LISVGAYIGLHDYNSENVETGAPVVIDpIDIGNQLAGINQSLQNAEDYIERSEEYLSGINPA I

[0367] SEQ ID NO : 193 , NinV (OQ438286)

[0368] MSENSHWYNIIYIIMKSRCSRLIGLIIVLTALMSAVVGIDFEGLSSIGVVKGR TLNYKIRGNSAYKILVIKLTPTIRSTSGAVNFENCTKDQISAHKVLVRNVLTPVKDALTS MKSKVTDYTPNSRLFGAIVAGAALVTATAATITAGVALHQSNQNAQAIANMKDAISKT NQAVSELKQGTQKLATVVDSLQNQINSQIIPVLNQLGCQTTGLTIGLYLTRYYSEILTVFG PAIQDPVNSALTIQAISKAFEGNFDKLMSAMGYSVSDLKDVLESDLVRGKIIDVDPDVGY MALQIEFPQITPVTGAHIQELLPISFNYKGDEWITILPSFVLNRMTYLSNIDIAHCLVTETS VICDNDLAMPMSTQMRDCLLGNTSKCSREQVITSYVPRFALSGGVIYANCLNTQCACAT NGRSISQSSRQTIMMLDDKDCKIYEVGGIFISVSTYMGIGKFENENISIGPPVVVDpIDVSGQLAEVNQSLSKTEELLEESNEYLSQVHVSLEXAMPLES

[0369] The Examples / Methods have been included to provide guidance to one of ordinary skill in the art for practicing representative embodiments of the presently disclosed subject matter. In light of the present disclosure and the general level of skill in the art, those of skill can appreciate that the following Examples are intended to be exemplary only and that numerous changes, modifications, and alterations can be employed without departing from the scope of the presently disclosed subject matter. The following Examples are offered by way of illustration and not by way of limitation.

[0370] Phylogenetic Analysis

[0371] Candidate sequences were identified by using the NCBI Protein BLAST tool (66). Nipah-Malaysia sequence NC_002728 was used as an input sequence. Sequences were aligned using the MAFFT algorithm (67). We removed any sequences for which there were gaps in the amino acid sequence, ensuring each entry contained a full ectodomain protein sequence. The final tree was built as a UPGMA based on the distance matrix using the Tree Viewer software (68). Alignments are shown in (Figures. 8-9).22F5 Antibody VH Chain (SEQ ID NO: 68) QLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPNNGGR NYVQKFQGRVTMTRDTSISTAYMELSSLSSDDTAVYYCARAGTSDWYFDLWGRGTLV TVSSAS22F5 Antibody VL Chain (SEQ ID NO: 69) DIVITQSPSSLAVSVGEKVTMSCKSSQSLLNSRTRRNYLAWYQQKPGQPPKLLIYWASTR ESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCKQSYNSWTFGGGTKLEIKRT

[0372] Protein Production

[0373] For HNV proteins, human codon-optimized DNA sequences were synthesized by Gene Immune Biotechnology and cloned into a paH plasmid vector. Fusion protein ectodomainsequences were followed at the C-terminal end by a foldon trimerization domain, HRV3C protease site, 8x his tag, and a twin-strep tag (IBA Lifesciences). Attachment protein head domains had an artificial secretion signal, secrecon (43) added to the N-terminal end and an 8x his tag added to the C-terminal end. HNV protein plasmids were transiently transfected into HEK 293 cell lines, either Expi293 or 293F (Thermo Fisher Scientific). The cells were allowed to incubate at 37°C, 8% CO2 and the supernatant was harvested 5 days post-transfection.

[0374] Fusion proteins were purified by first sequestering free biotin by applying BioLock blocking solution (IBA) and then using the Strep-Tactin affinity chromatography resin (IBA). Eluted protein was then further purified using a Superose 6 Increase 10 / 300 GL size-exclusion column (Cytiva) with PBS buffer at pH 8. The elution from SEC was spin concentrated with lOOkDa filters and flash frozen with liquid nitrogen.

[0375] Attachment protein head domains were purified first by sequestering EDTA by adding 2 mg of Cobalt Chloride per IL of supernatant. TALON Cobalt Resin (Takara) was used for purification, with 20 mM HEPES and 150 mM NaCl at pH 8 as a buffer, adding 50 mM Imidazole for elution. Proteins were further purified with a Superdex 75 Increase 10 / 300 GL SEC column (Cytiva), again using the HEPES-NaCl buffer. The elution from SEC was spin concentrated with 10 kDa filters and flash frozen with liquid nitrogen.

[0376] Antibody human codon-optimized DNA sequences were synthesized by Gene Immune Biotechnology and cloned into pVRC8400 plasmid vectors. Heavy chains were designed to begin with the beginning of the antibody sequence and continue through the human IgGl Fc region. Light chains included both the VL and CL domains. Depending on the sequence source, some antibody sequences either utilized mouse or human VL and VH domains. Mouse anti-NiV-F fabs were designed to end after the mouse CHI domain and had an 8x his tag added to the C-terminal end (38).

[0377] Plasmids encoding the heavy and light chains of the antibodies IgG 1E5, nAH1.3, 4G5, and DH851.3 were co-transfected into Expi293 cells at a 1: 1 ratio, using the ExpiFectamine 293 Transfection Kit (Thermo Fisher Scientific, Cat. No. A14525), according to the manufacturer's instructions. Following a six-day incubation period post-transfection, the cellswere harvested, and the culture supernatant was clarified by filtration through a 0.2 pm PES membrane filter to remove cellular debris. IgG purification was subsequently carried out using Protein A affinity chromatography. The filtered supernatant was passed through the Protein A affinity column (Pierce™ Recombinant Protein A Agarose Cat No. 20334), and the bound IgG was eluted using IgG elution buffer by adjusting pH via Tris Base. Eluted fractions were pooled and concentrated with Centricon-70 30 kDA filter (Millipore Sigma) followed by size-exclusion chromatography using a HiLoad 16 / 600 Superdex 200 pg column (Cytiva), pre-equilibrated with l x phosphate-buffered saline (PBS, pH 7.4) containing 0.02% sodium azide to prevent microbial contamination. The purified IgG was collected, concentrated, and analyzed for purity and integrity by SDS-PAGE and Tycho NT.6 (NanoTemper Technologies) for Differential scanning fluorimetry (DSF) assay.

[0378] Anti-NiV-F fabs were purified using the same Cobalt resin-based procedure as described above for head domains.

[0379] To generate Fab fragments for 22F5 mouse IgG and 1C8 mouse IgG, 2 mg of each IgGs were passed through Zeba Spin Desalting Columns (ThermoFisher), then washed with digestion buffer containing 35 mg cysteine. IgGs were incubated with 0.250 ml of the 50% slurry of Immobilized Papain resin (ThermoFisher) followed by incubation with digestion buffer containing 35 mg cysteine at 37 °C overnight in microcentrifuge tubes. The digested Fabs were separated from the Immobilized Papain by centrifuging at 5000 x g for 1 minute, followed by washing with digestion buffer. After digestion, the mixture was subjected to protein A affinity chromatography (ThermoFisher) then monitored by SDS-PAGE to confirm Fab generation. To further purify the Fab fragments, Size-Exclusion Chromatography (SEC) was used for purification of the Fab fragments using Superose 6 Increase 10 / 300 column GL (Cytiva). The sample was loaded onto the column at a volume of 0.5 mL and the elution was carried out with PBS at a flow rate of 0.5 mL / min. Fractions were collected, yielding a final concentration of 3.2 mg / mL and 4.2 mg / ml for 22F5 Fab and 1C8 Fab, respectively. The purified Fab was stored at -80°C. The ELISA control antibody Ab82 was purified as described previously (57).

[0380] Differential scanning fluorimetry (DSF)

[0381] Samples were analyzed using a Prometheus Panta (Nanotemper Technologies). All samples were diluted to 0.125mg / mL using PBS, pH 8. Roughly 10 pL was added to glass capillaries in triplicate for measurements. The instrument was programmed to measure 350nm and 330nm fluorescence from 35.0-95.0°C using a 6.0°C / min temperature gradient. All curves reported represent averages from the triplicate readings.

[0382] Bio-layer interferometry (BLI) was employed to assess the binding between fusion proteins and murine monoclonal antibodies (mAbs) or fragment antigen-binding regions (Fabs). The analysis was performed using an Octet RH16 (Octet RED384) system (Sartorius) at 25°C with kinetics buffer (Sartorius) as the running buffer. HNV-F proteins with the twin-strep tag at 6 pg / mL were captured onto Streptavidin (SA) Biosensors (Sartorius), followed by a 60-second wash in kinetics buffer. The biosensors were then exposed to 20 pg / mL mAbs or 50 pg / mL Fabs for 300 seconds to allow for association with the fusion protein. Dissociation in kinetics buffer was monitored for an additional 300 seconds. Sensor data was analyzed using Octet Analysis Studio (Sartorius), with reference biosensor and zero-analyte control sensorgrams subtracted from the raw data to obtain the curves corresponding to specific binding events.

[0383] For additional fusion protein experiments (Data not shown), the analysis was performed with the same equipment, tips, and buffer. HNV-F proteins were loaded at 6 pg / mL for 300s and equilibrated in buffer for 60s before a 120s association step and 200s dissociation step. All analytes were at 50pg / mL. Regeneration with lOmM Glycine, pH 1.7 was employed, using tips for 6 cycles and regenerating with 10s pulses between regeneration solution and buffer. SARS-S2-HexaPro 70 was loaded onto a tip as a reference sensor. Kinetics buffer was used as a reference sample. Double reference subtraction was performed, except for in the case of Griffithsin IgG, which bound to the SARS-S2-HexaPro, so only reference sample subtraction was used. For each cycle, NiVop08 was loaded and bound to 4B8 Fab, and the signal relative to the first use of the tip was used to adjust the response values of all association curves during that cycle. For attachment head domains with human Fc-containing Abs, the analysis was performed using an Octet RH16 (Octet RED384) system (Sartorius) at 30°C with HBS-EP+ Buffer (Cytiva) as the running buffer. Antibodies at 2 pg / mL were loaded onto anti-human antibody capture (AHC)biosensors (Sartorius) for 300s, followed by a 60s buffer baseline. Association with head domains at 25 pg / mL lasted 120s, followed by dissociation in buffer for 200s. Between cycles, biosensors were regenerated by alternating 3x in 1 OmM Glycine pH 1.0 buffer and running buffer for 5s each. Sensors were used for no more than 10 cycles of regeneration. Double reference subtraction was performed using both a control-loaded sensor and a zero-analyte reference sample.

[0384] Surface Plasmon Resonance

[0385] Binding and kinetic assays were perfomed by Surface Plasmon Resonance (SPR) on a T-200 Biacore system (Cytiva) at 25°C. All the binding and kinetic experiments were carried out in the HBS-EP+ 778 (10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA and 0.05% surfactant P-20) running buffer. For the binding assay, anti-Fc mouse SPR chip were used to capture the 200nM of 22F5 IgG at lOul / min flow rate and 50nM of LayV-l-F ectodomains as analyte were run at 30ul / min flow rate and association and dissociation were monitor at 120 seconds. In the competitive assay, 200nM of 4G5 IgG was captured on anti-Fc human SPR chip at 10 pl / min and analyte as 22F5 Fab with Lay V-l-F ectodomain complex in 5: 1 ratio was flowed at 30 pl / min flow rate for 60 seconds. SPR SA chip was used to capture the 50 nM of the LayV-l-F ectodomains at 5 pl / min flow rate by biotin tag for kinetics analysis. 22F5 Fab 50 nM with two-fold dilution (50 nM, 25 nM, 12.5 nM, 6.25 nM and 3.12 nM) as analyte was flowed with 50 pl / min flow rate and association and dissociation time were analyzed at 60 and 240 seconds, 8 respectively. For the reference curves, we kept the same parameter in reference flow cells without protein capture. The sensorgrams were blank corrected and analyzed in the Biacore T- 200 evaluation software.

[0386] For Ephrin-HNV-G interactions, Binding experiments were performed on a Biacore T-200 (Cytiva) with HBS buffer supplemented with 3 mM EDTA and 0.05% surfactant P-20 (HBS- EP+, Cytiva, MA). All binding assays were performed at 25°C. Ephrin binding to head domains was assessed using a Series S CM5 chip (Cytiva, MA), which was labeled with antihuman IgG (Fc) antibody using a Human Antibody Capture Kit (Cytiva, MA). Ephrin was coated onto the chip at 12.5 nM (120 s at 5 pL / min). Head domains were injected at 100 nM over the ephrin using a single injection (60 s on time, 180 s off time at 50 pL / min). The surface was regenerated with 3 pulses of a 3 M MgC12 solution for 10 s at 100 pL / min.

[0387] Negative-Stain Electron Microscopy

[0388] Frozen samples from -80°C were thawed at room temperature in for 5 minutes. Samples were then diluted to 40 pg / mL with 0.02 g / dL Ruthenium Red in HBS (20 mM HEPES, 150 mM NaCl pH 7.4) buffer containing 8 mM glutaraldehyde. After 5-minute incubation, glutaraldehyde was quenched by adding sufficient IM Tris stock, pH 7.4, to give 80 mM final Tris concentration and incubated for 5 minutes. Quenched sample was applied to a glow- discharged carbon-coated EM grid for 10-12 seconds, blotted, consecutively rinsed with 2 drops of 1 / 20X HBS, and stained with 2 g / dL uranyl formate for 1 minute, blotted and air-dried. Grids were examined on a Philips EM420 electron microscope operating at 120 kV and nominal magnification of 49,000x, and images were collected on a 76 Mpix CCD camera at 2.4 A / pixel. Images were analyzed by 2D class averages using standard protocols with Relion 3.0 (58).

[0389] Mass Photometry

[0390] A TwoMP (Refeyn) mass photometer was used for analysis. Calibrated system with a three-point curve consisting of [3-amylase, Apoferritin, and Thyroglobulin. Samples were diluted to 40 pM in PBS. To each well, lOpL of PBS was added to focus the system before adding 10 pL of sample and mixing for a final concentration of 20 pM. Recorded data for 60s, with the machine counting individual adsorption events. The DiscoverMP processing software (Refeyn) was used to assign molecular weights to individual events.

[0391] Methods for Mouse Immunizations

[0392] Immunization in VH1-2RJH2 / VK1 -33RCSA hTdT(SE13) mice (59) were intramuscularly immunized with Lay V-F_WT (25 mcg / animal) and NiVop8 (25 mcg / animal) adjuvanted with GLA-SE (IDRI EM-082, 5 mcg / animal) at weeks 0, 4, 15 and 21. Serum titers were monitored by ELISA as described below. Mice with high-binding antibody titers were selected for the subsequent spleen cell fusion and B-cell sorting experiments.

[0393] Hybridoma cell line generation and monoclonal antibody production

[0394] Mice were boosted with the indicated priming antigen 3 days prior to fusion. Spleen cells were harvested and fused with NS0 murine myeloma cells using PEG1500 to generate hybridomas (37). After 2 weeks, supernatant of hybridoma clones were collected and screenedby binding ELISA as described below. Hybridomas that secreted Lay V-F reactive antibodies were cloned by limiting dilution until the phenotypes of all limiting dilution wells were identical. IgG mAbs were purified by protein G purification.

[0395] Indirect binding ELISA method

[0396] ELISA assays were utilized for the purposes of measuring mouse serum antibody titers, screening for antigen specific hybridoma clones and characterizing the binding of monoclonal antibodies. 384 well ELISA plates (Costar #3700) were coated with 2 mcg / ml streptavidin (Thermo Fisher Scientific Inc. Cat. No. S-888) or protein antigen in 0.1 M sodium bicarbonate overnight at 4°C. Plates were washed with PBS / 0.1% Tween-20 and blocked for one hour with assay diluent (PBS containing 4%(w / v) whey protein / 15% Normal Goat Serum / 0.5% Tween-20 / 0.05% Sodium Azide). Streptavidin coated plates were washed and followed by 10 pl Twin-Strep-tag® (IBA Lifesciences GmbH) protein at 2 mcg / ml in assay diluent for one hour. All plates were then washed and samples were added in 10 pl volumes depending on sample type as follows. Mouse serum was diluted 1 to 30 and titrated three-fold. Hybridoma supernatants were added undiluted as a single well. Monoclonal Fabs and IgG were added starting at 100 mcg / ml in three-fold titration. Plates were washed and lOpl goat anti-mouse IgG-HRP secondary antibody diluted 1 :10,000 (Southern Biotech #1030-05) (for serum and hybridoma supernatants), goat anti-human IgG Fab-HRP diluted 1: 15,000 (Jackson Immunoresearch, 109-035-097 ) or goat anti-Mouse IgG Fab-HRP diluted 1 : 10,000 (Southern Biotech 1015-05) (for monoclonal Fabs and IgG) in assay diluent without azide was incubated for 1 hour, washed again and detected with 20 pl SureBlue Reserve (Seracare 5120-0081) for 15 minutes. Reaction was stopped with the addition of 20 pl HCL stop solution. Plates were read at 450 nm using a Spectramax 384 plus.

[0397] Antibody Gene Cloning and Sequencing

[0398] Cell suspensions of the 22F5 hybridoma cell line were pelleted by centrifugation at 300 g for 5 minutes. The cell pellets were washed twice with Phosphate-buffered saline (PBS, IX) to remove residual culture medium. The 22F5 antibody genes were amplified from the hybridoma cells by RT-PCR. Briefly, RNA was isolated from the cell pellets by adding them toreverse transcription solution. Immunoglobulin genes were reverse transcribed using Superscript III (Thermo Fisher Scientific) with random hexamer oligonucleotides (Gene Link, Hawthorne, NY) as primers. The cDNA served as a template for nested PCR amplification of the antibody heavy and light chain genes. The PCR used mouse antibody gene-specific primers 62 and was carried out with AmpliTaq Gold 360 (Thermo Fisher Scientific). The PCR products were purified using the Biomek FX Laboratory Automation Workstation (Beckman Coulter, Brea, CA). Purified PCR products were then sequenced using Sanger sequencing to obtain the antibody gene sequences. The V(D)I rearrangement, somatic hypermutation frequency, and CDR3 (complementarity-determining region 3) length of the VH (heavy chain variable region) and VK / L (light chain variable region) genes were analyzed using a custom in-house bioinformatics pipeline for immunogenetic analysis. Annotation was performed using Partis (74) with a custom knock-in mouse reference which combined the knocked-in human VH1-2*O2, JH2*01, and VKl-33*01 with mouse endogenous V, D and J gene segments.

[0399] Cryo-EM grid Preparation and Imaging

[0400] To prepare grids, UltaFoil Rl.2 / 1.2 (Cu, 300-mesh; Electron Microscopy Sciences, PA) grids were glow discharged for 15 seconds at 15 mA using a PELCO easiGlow Cleaning System (Ted Pella Inc). The final concentration of purified AngV-F protein was maintained as 1.5 mg / ml in PBS. and to create air-water interface during the vitrification, samples were incubated with. 0.005 % (w / v) of n-dodecyl P-D-maltoside (DDM) was added to the protein samples before applying to the grid to prevent the protein form adhering to the air-water interface. 3.0 pL of the protein sample was applied to the grid and incubated for 30 seconds at >95% humidity. Excess protein was blotted away for 2.5 seconds before being plunge frozen into liquid ethane using Leica EM GP2 plunge freezer (Leica Microsystems). Cryo-grids were imaged using a FEI Titan Krios (Thermo Scientific) or Tundra cryo TEM (Thermo Scientific).

[0401] The Titan Krios TEM was used for the determination of the AngV-F trimer, dimer-of- trimer, and 22F5 Fab-LayV-l-F complex structures. The Krios was equipped with K3 camera (Gatan) at 81k magnification, operated at 300kV. For AngV-F, approximately 24,000 micrographs were collected at nominal defocused range between -2.8 to -1.7 pm with a doserange of 63-65 e / A2. For LayV-l-F, 9,715 micrographs were collected at a nominal defocused range between -2.8 to -1.2 pm with a dose range of 58.6-60.6 e / A2.

[0402] The Tundra TEM was used for determination of the AngV-F hexameric lattice and 22F5 Fab-1066 LayV-F-DS complex structures. For the AngV-F hexameric lattice, theTundra was equipped with CETA-F camera (Thermo Scientific™) at 180k magnification, operated at lOOkV. Approximately 4,000 micrographs were collected at nominal defocused range between - 3.0 to -1.5 pm with dose range of 23-25 e / A2. For the 22F5 Fab-LayV-F-DS complex, the Tundra was equipped with a Falcon 4i camera (Thermo Scientific) at 180k magnification, operated at lOOkeV. 9,514 micrographs were collected at a nominal defocused range between - 2.5 to -0.7 1pm with a dose of 30.72 e / A2.

[0403] Cryo-EM Data Processing

[0404] Cryo-EM image quality was monitored on-the-fly during data collection using automated processing routines. Data processing was carried out using cryoSPARC (60, 61). Micrographs were curated through contrast transfer function (CTF) where greater than >30 A were discarded. Automated blob picker was used to assign the particle position. Different box sizes are utilized to accommodate the varying dimensions of specific molecular assemblies. For instance, monomer particles are extracted using a box size of 320 pixels, while dimer particles require a larger box size of 512 pixels. For even larger molecular assemblies, such as hexamer lattices, a box size of 1024 pixels was employed. Following particle extraction, multiple rounds of 2D classification was performed to remove junk, ab-initio 3D reconstruction was used to create 3D reconstructions and poor-quality particles were discarded after through heterogenous refinement. Final resulting volumes were subjected to non-uniform refinement to build a high- resolution 3D reconstruction. Phenix (Macromolecular structure determination using x-rays, neutrons and electrons: recent developments in phenix (62), Coot (63), Pymol, nextPYP (64) and ChimeraX (65).

[0405] X-ray crystallography.

[0406] Crystals were grown via the vapor diffusion method at room temperature in siting drop well format. GakV-2-G head domain protein crystals were formed by mixing 0.20 pL ofprotein at 15 mg / mL with 0.20 piL of well solution containing 0.2 M magnesium sulfate heptahydrate, 20% w / v PEG3350 at pH 7.1 . Crystals appeared within one week.

[0407] Cryoprotection of the crystals was done by mixing glycerol to a final concentration of 50% with the well solution. Crystals were moved to the drop containing the well solution and glycerol and soaked for one minute then flash-cooled in liquid nitrogen and data were collected at the NSLS-II, Beamline 19-ID at 100 K and a wavelength of 1 A. All data were indexed and integrated using iMosflm (66) and scaled using AIMLESS.

[0408] Phaser (67) in the PHENIX suite was used to perform molecular replacement using a model generated from the AlphaFold3 prediction server (68) using the GakV-2-G head domain sequence. Phenix. refine (69) was used for data refinement, and manual refinement was done in Coot (63).EXAMPLE 1

[0409] Identification and Classification of Henipavirus Species

[0410] To begin categorizing this diverse set of Henipavirus strains, we identified as many unique Henipavirus G and F proteins as could be found, assessed the phylogeny of these sequences, and developed a universal naming scheme. The NCBI BLAST tool (21) was employed to search for all available unique sequences related to the input sequence (NC_002728) (23). In addition to utilizing BLAST, we carried out an extensive literature search to include other sequences that may not have been discoverable through BLAST, such as Gamak virus (7). After identifying related sequences, we narrowed the list to include only sequences from mostly complete genomes, leaving out those where only partial DNA sequences of only G or F proteins were available. Almost all the identified strains had genome lengths close to the canonical 18.2 kbp for Henipaviruses (24), with some species being slightly longer. No sequence in our panel had a deposited genome shorter than 11.2 kbp. The several sequences that were shorter than the rest were either Nipah-Malaysia or Hendra strains that were not recently discovered and have been well-characterized in the literature.

[0411] Next, the Clustal Omega tool (25) was used to align the G and F amino acid sequences. As our goal was to purify soluble ectodomain constructs, we truncated the sequences, using trends in the sequence alignments to decide on exact truncation points between species. All HNV-F ectodomains were of similar length, allowing all to be truncated at the C-terminal end of the ectodomain, at the site equivalent to Nipah virus residue 488 to be consistent with previous purifications of F ectodomains (19). For the G proteins, the boundaries of domains do not align as consistently across species as they do with the overall less variable F protein, though the N- terminal portion of the sequence up to the end of the transmembrane domain was relatively consistent across all species, allowing for the truncation position for the ectodomain to be set at the analogous site to Nipah virus residue 71.[00412J While the full G ectodomain and its elusive fusion promotion mechanism is an important area for research, in this study we focus on the G head domain that harbors the receptor binding site and is targeted by a myriad of neutralizing antibodies (14,16,41,48). Therefore, while our sequence and phylogenetic analysis considers the whole genome or full G amino acid sequence, only the head domains of G proteins were purified. For these head domains, sequence variability resulted in the transition between the neck domain and head domain being less clear from sequence. In classical Henipavirus sequences, there are frequent glycine and proline residues leading up to a conserved cysteine in the head domain. The Parahenipaviruses also contain a proline-rich region before the head domain, with as many as five consecutive prolines in some sequences. This proline-rich region is often followed in Parahenipaviruses by many aspartic and glutamic acid residues. For G Head domains, the site analogous to Nipah virus residue 177 was selected as a suitable start point based on sequence analysis and available structural data for Nipah and Langya G proteins (14, 26). After removing redundant sequences where differences were noted only in the signal sequence, transmembrane domains, cytosolic domains, or other regions not included in the final F ectodomain or G head domain constructs, the panel was assembled into a phylogenetic tree based on complete genomes (Figure 1). From this list, a subset of unique F ectodomains and G head domains was purified that would broadly sample the diversity of species in the phylogenetic tree, resulting in 35purified F ectodomains and 32 purified G head domains. The exact amino acid sequences for the F and G proteins can be found in Figures 8 and 9.

[0413] The International Committee on Taxonomy of Viruses (ICTV) has recently updated terminology related to the Henipaviruses, establishing the Henipavirus and Parahenipavirus genera, and specifying select species within these groups (3). Our effort to compare the properties of dozens of unique F and G sequences required finer granularity than the species level, and as a result, for this study we adopted an abbreviation and numbering scheme to aid in distinguishing specific sequences. The basis for this terminology is expanded upon in our description below and the corresponding accession code and current ICTV name for each sequence is listed in Figure 10.

[0414] For clarity in referring to specific sequences with greater granularity than the species level, we based our abbreviations and numbering on conventions of Nipah virus naming in the literature. Previously, strains have been named based on the location where the sequence was reported, specifically, Bangladesh (NiV-B) and Malaysia (NiV-M). Recent discovery and classification of new Nipah virus strains support the existence of potentially two further geographical groupings, India (NiV-I) and Cambodia (NiV-C) (57,58), and these distinctions are apparent in our tree as well. With the Bangladesh strains, the phylogeny revealed the grouping of strains into two clades, prompting us to denote the two groupings as NiV-B 1 and NiV-B2, respectively. From here, unique entries were designated an additional number to further classify the variant (i.e. NiV-B2.1, NiV-B2.2, NiV-B3.1, etc.). An identical scheme was applied for the India strains, which previously have been referred to as members of the Bangladesh clade (57?). Malaysia strains were classified by number as well. Entry MK801755 was identified during surveillance in Cambodia in 2003, where it was isolated from Pteropus lylei fruit bats (59), and as the only unique NiV-C member of the panel, requires no numbering.

[0415] Previous studies have established two clades for Hendra virus (29), in which the more recently discovered clade is referred to as HeV-g2. However, due to the use of G as an abbreviation for the attachment protein, we sought to adopt a different naming scheme to avoid confusion. Unlike Nipah, where genomic differences align with country borders, all Hendra virusdiscoveries detections have been in Australia. Therefore, the two distinct clades have been named Alpha (a) and Beta (0), with each member of the clade given a number. The other Henipaviruses that have emerged from a fruit bat reservoir (Figure 1) have all previously been named in the literature. These include Cedar Virus (CedV)(30), Ghana Virus (GhV)(31), and Angavokely Virus (AngV)(32). Our panel includes two unique Cedar virus attachment proteins; therefore, these strains were numbered 1 and 2. There are only singular strains of Ghana and Angavokely viruses identified, however some evidence indicated that the originally deposited sequence for Ghana virus F protein may have included an inaccurate N-terminal end of the protein, and a corrected sequence was indicated (33). While our phylogenetic tree only includes the one strain reported, our panel of purified proteins included both F protein sequences.

[0416] Among Henipaviruses found in shrews, now officially classified as members of the Parahenipavirus genus, several already have established names in the literature, including Mojiang Virus (MojV), Langya Virus (LayV), Gamak Virus (GakV), Denwin Virus (DewV), Melian Virus (MelV), and Ninorex Virus (NinV) (2, 7, 8, 34). For those with more than one strain reported, the same numbering approach used with the Nipah-Bangladesh clade was used. In the case of Langya and Daeryong viruses, distinct clades are found with geographic separation, as with Nipah virus. Similarly, these clades were assigned as Korea and China for clarity.

[0417] Many of the recently discovered Parahenipavirus sequences have been given repetitive names based on their shared collection site and host species. However, in some situations, this could imply overly close or distant phylogenetic relationships between strains. For example, strains listed as Jingmen Crocidura shantungensis virus 1 (e.g. OM030314) (9?) are not closely related to those listed as Jingmen Crocidura shantungensis virus 2 (e.g. OM030315). Inversely, strains listed as Wenzhou shrew Henipavirus 1 (OQ715593.1) and Wenzhou Apodemus agrarius Henipavirus 1 (MZ328275.1) were shown to be quite closely related (Figure 1). For simplicity and clarity, we grouped many of these sequences using variants of the name “Shrew Henipavirus (SHNV)” with a number, thus establishing SHNV1, SHNV2, SHNV3, SHNV4, through SHNV11. For SHNV5 and SHNVIOspecifically, there were several strains thatthe phylogenetic tree indicated were closely related enough to be considered the same species (Figure 1). As a result, we utilized a numbering scheme as described above.

[0418] Lee et al., 2020 (7), previously reported the discovery of Daeryong Virus in Korea, our phylogenetic analysis revealed that Jingmen Crocidura shantungensis virus 2 (OM030315, PP272750.1)), detected in China, could be considered the same species as Daeryong Virus. Following the naming procedure used for Nipah virus, the strains were given the names Daeryong virus-Korea (DarV-K) and Daeryong virus-China (DarV-C).This demonstrates that several of these newly discovered Parahenipaviruses are geographically well- distributed.

[0419] For F proteins, the sequence identity relative to NiV-Ml-F is high for both NiV and HeV strains, with all NiV sequences over 98% and HeV strains over 97%. The sequence identity of most non-NiV / HeV strains to NiV-Ml-F is -40%, with sequence similarity around 55-60%, revealing that the F protein is relatively well conserved across genera. A phylogenetic tree of HNV-F proteins based on amino acid sequences was overall similar to the whole genome tree with several small differences. Specifically, the distinction between NiV-C-F and NiV-M strains is more apparent in the F-based phylogenetic tree, the F proteins of SFTNV3 and SHNV11 appear to be very closely related, and AngV-F appeared distinct from all other HNV strains, rather than part of the bat clade as seen through full-genome phylogeny.

[0420] Compared to F proteins, HNV-G proteins demonstrate much greater sequence variability. Sequence identity values are only -78% between NiV and HeV, and drop to below 20% for Parahenipavirus G proteins, compared to NiV-Ml-G. The sequence similarity scores are also rather low, with most Parahenipavirus strains at around 35% similarity to NiV. When considering amino acid-based phylogeny, the relationships between strains remain similar as we had observed for the F ectodomains. One difference is that SHNV3-G and SHNV11-G, as with their F counterparts, appear more closely related than their full genome indicates. Overall, the limited sequence similarity observed for the G head domains suggests that there could be greater antigenic, structural, and mechanistic differences between different HNV-G proteins than among F proteins.EXAMPLE 2

[0421] Preparation of diverse Henipavirus F ectodomains

[0422] We expresssed 35 HNV-F constructs that encoded the ectodomains spanning residues 1 to the equivalent of 488 in NiV, followed by a foldon trimerization domain (35) and a C- terminal Twin Strep tag. For all constructs, the putative natural cleavage site that allows processing from the F0 204 precursor to the fusion-competent F1 / F2 complex was retained. The proteins were expressed through transient transfection in 293F cells and purified using their C- terminal Strep affinity tags followed by size exclusion chromatography (SEC). We obtained variable yields of F proteins, with some constructs expressing over 2 mg / L and others less than 0.1 mg / L. For eight F proteins, the yields were too low to perform any downstream studies. We found that yields could vary between strains that only differed by a few amino acids. For example, NiV- B 1.1 -F and NiV-B 1 ,2-F differ by only three residues in the ectodomain, but while NiV-B 1 ,2-F produced ~1 mg / L, NiV-B 1.1 -F had no yield across two purification attempts.

[0423] HNV-F ectodomains typically elute from SEC with three peaks, one that eluted immediately after the column void volume, a broad “middle peak”, and the main peak that corresponds to the molecular weight of an F trimer eluting last (Figure 2A), with the ratio of the peaks to one another differing from strain to strain. Furthermore, the middle peak of some strains can sometimes be seen to have a shoulder, indicating there could be multiple sized components eluting within this peak. The SDS-PAGE profile of the middle peak fractions showed a major band at the molecular weight expected for the trimeric ectodomain unit, indicating that the presence of the higher molecular weight peak was not a result of any covalent linkage or differential glycosylation that would change the molecular weight enough to be distinguishable on SDS-PAGE . Both peaks individually or mixed together generated an essentially identical molecular weight profile, with each being dominated by the expected molecular weight of the trimer, and both showing a very small presence of species twice to three times the weight of the trimer (Figure 2A, insert). SDS-PAGE also indicated that all F proteins are being expressed in their Fo state, without any proteolytic processing to F1 / F2. Negative stain electron microscopy (NSEM) of each middle or main peak reveals comparable sets of two-dimensional class averages. Thetypical pre- and postfusion shapes identified previously (19) can be seen throughout the panel for both middle and main peak components. Taken together, these data suggest that the middle peak is composed of transient, non-covalently associated F ectodomain trimers, which are reminiscent of previous studies of Paramyxoviridae F proteins where oligomeric states were reported for the F protein (41,75,39). In summary, though the F protein expression yields varied greatly, similar profiles were observed through SEC, SDS-PAGE, and NSEM analysis across all HNV species tested.EXAMPLE 3

[0424] Antigenicity of HNV F ectodomains

[0425] We next characterized the antigenicity of the F ectodomain panel. Since many new HNV species have been recently identified and given the limited availability of antibody sequences with known reactivity to LayV and other newly categorized members of our panel, we sought to identify antibodies of novel specificities by vaccination of mice with HNV-F antigens (Figure 2B). One group was given four injections of the LayV-Cl-F ectodomain, and for two others, the fourth injection of LayV-Cl-F was replaced with NiV, either the wild-type NiV-Ml- F, or NiVop08, a pre-fusion stabilized NiV construct (36). By boosting with NiV constructs, we sought to elicit broadly reactive antibodies. Using hybridoma fusion technology (37) we identified two murine IgGl antibodies, named 1C8 and 22F5, both from group 2, with reactivity to LayV-l-F. An additional vaccination series, using the same antigens with a similar timeline, led to the isolation of an additional four murine antibodies, all from group 2, named 9A9, 8C7, 20G7, and 8D4.

[0426] The binding of antibodies elicited in our immunization experiments, as well as previously identified anti-NiV or anti-LayV antibodies, to our panel of HNV-F ectodomains was assessed by enzyme-linked immunosorbent assay (ELISA), surface plasmon resonance (SPR), or Bio- layer interferometry (BLI) (Figure 2C) (20, 38). 1H1 and 4B8, previously identified as anti-NiV-F antibodies, displayed broad reactivity to all NiV and HeV strains, but extremely limited reactivity to all other species. A panel of previously characterized mouse anti-NiV-FFabs broadly reacted to nearly all NiV and HeV antigens, as expected. 4G5, previously identified as an anti-LayV-F and Moj V-F antibody that binds both the pre- and post- fusion conformations at an epitope involving the HRA and HRB regions, not only strongly bound to LayV-F and MojV-F, but also to SHNV5-1-F and SHNV5-2-F, thus indicating that antibodies targeting the 4G5 epitope have greater breadth than was previously known (Figure 2C).

[0427] Of the antibodies isolated from the immunization studies, 8C7, 20G7, and 9A9 bound only to NiV-F ectodomains. 1C8, 22F5, and 8D4 demonstrated strong reactivity not only to 268 LayV-F proteins, but also to MojV-F and both SHNV5 strains. Moreover, 22F5, which generated stronger binding signals than 1C8, bound to SHNV2-F as well (Figure 2C). 8D4 demonstrated the broadest binding profile, with binding detected to F proteins from each of GakV-1, GakV-2, NinV, SHNV1, SHNV2, and SHNV3. Considering the phylogenetic relationships of these strains (Figure 1), 8D4’s reactivity may cover nearly the entirety of the known Parahenipaviruses. However, 8D4 demonstrated sharply reduced binding to pre-fusion stabilized F constructs, indicating that it specifically bound to the post-fusion F conformation. Since there are very few well-characterized antibodies that have been described for Parahenipaviruses, we selected 22F5 for detailed characterization, given its combination of a broad cross-reactivity profile and ability to bind pre-fusion F proteins.

[0428] To assess the potential to target the HNV-F protein glycans, we tested the binding of 2G12, a glycan reactive antibody previously noted for its broad specificity to diverse viral fusion proteins and ability to neutralize influenza and HIV-1 (76-77). We did not detect any binding between 2G12 and the HNV-F panel. We also tested the reactivity of recombinant Griffithsin, a lectin discovered in Griffithsia red algae that has been investigated for antiviral properties (78-79) and has been shown to be protective against NiV challenge in hamsters (80). Griffithsin bound to most of the NiV / HeV strains in the panel, while also binding to two prefusion stabilized Parahenipavirus constructs, of SHN5-1 and GakV-1, specifically, underscoring the potential for a glycan-directed approach for broad spectrum targeting of HNVs.

[0429] Using (SPR), we tested the binding of 22F5 to LayV-F ectodomains in various conformational states, using two separate disulfide stabilized pre-fusion LayV-F constructsdescribed previously, that we refer to here as DS (39) and DSv2 (20). Additionally, we tested binding with and without competition with 4G5 (Figure 2D). 22F5 bound to Lay V-F in both pre- and post-fusion conformations, indicating conformational bispecificity, and its binding was mostly unaffected by the presence of 4G5, indicating a distinct epitope. Given the strong and diverse binding profile of 22F5, we selected it for further characterization, including sequencing and structure determination.

[0430] Taken together, our findings align with the current understanding that Henipaviruses and Parahenipaviruses have distinct antigenic profiles. Parahenipavirus F proteins are not bound by known anti-NiV / HeV antibodies. However, the breadth of strains bound by our LayV-F-reactive antibodies indicates that cross-neutralization of a sizeable portion of known Parahenipaviruses with a small number of antibodies can be achievable.EXAMPLE 4

[0431] Structural Characterization of 22F5 Antibody

[0432] Using single particle cryo-EM, we determined a structure of the 22F5 Fab bound to the post-fusion form of the LayV-Cl-F protein, at a global resolution of 4.5 A (Figure 3A), with local resolution of ~3 Aat the interface of 22F5 with the F protein. The 22F5 Fab bound to the DII region (11) of Lay V-F, with a few DI contacts as well. While there have been several previously observed antibodies targeting DII epitopes in NiV-F and HeV-F proteins (38), the only other available structure for an anti-LayV-F antibody, 4G5, details an epitope that includes contributions from HRA and HRB (20). Additionally, we determined the structure of the 22F5 Fab bound to the LayV-F-DS ectodomain at ~4A local resolution at the interface (Figure 3B), confirming that the 22F5 epitope is accessible in both the pre- and post-fusion conformation of LayV-F. For both LayV-Cl-F and LayV-F-DS, 22F5 was seen bound at all three DII sites.

[0433] The 22F5 Fab interacts in a mostly conserved manner with both the pre- and postfusion LayV-F ectodomains, utilizing both its heavy and light chains. The LI, L3, and H2 complimentary-determining regions (CDR) are particularly involved (Figure 3C). One notable change to the epitope between the pre- and post-fusion states is the lack of contact with thefusion peptide in the post-fusion conformation (Figure 3C, bottom). 22F5 binding appears to heavily rely on polar interactions (Figure 3D). Specifically, heavy chain residues N54 and R57 contact LayV-F DII residues N384 and the main chain of Q398, respectively. In the light chain, R27, Y32, and Y92 interact with DII residues D391, D393, and Q416, respectively. Additionally, the 22F5 light chain N terminal residue DI interacts with N285, a residue that belongs to domain DI of the neighboring protomer. When bound to the pre-fusion conformation, 22F5 contacts the neighboring protomer DI via its CDRH2 and framework region residues. Due to the shifting of protomers relative to each other during pre- to post-fusion conformational conversion, 22F5 shifts its interaction with the neighboring protomer DI region, losing its CDRH2 and heavy chain framework region contacts and gaining the light chain DI contact with the post-fusion F protein. (Figure 3C and 3D). Using SPR, we determined the affinity of 22F5 for both ectodomain constructs, as well as a heat-treated sample of the wild-type LayV-Cl-F that ensured uniform postfusion conformation, with binding observed at low nM affinities in all cases (Figure 3E). Interestingly, despite the loss of contacts with the fusion peptide residues, when bound to the post-fusion conformation of LayV-F, 22F5 bound tighter to the post-fusion conformation than the pre-fusion conformation. Given that the fusion peptide is a flexible region, it is possible that contact with it is entropically unfavorable, thereby explaining the improved affinity with the post-fusion LayV-F where this contact was absent.

[0434] In summary, 22F5 binding and structural data describe an epitope that is accessible in both pre-and post-fusion conformations and reveal a region on the Parahenipavirus F protein that can be targeted for greater breadth than previously recognized. 22F5 is a valuable addition to the antibody toolbox for the Parahenipavirus genus for which relatively few antibodies are known.EXAMPLE 5

[0435] Differential Scanning Fluorimetry ofHNV F ectodomains

[0436] We assessed the stability of the purified F ectodomainsusing Differential Scanning Fluorimetry (DSF), a label-free method that measures changes in intrinsic fluorescence as the proteins unfold. The DSF profiles of the two wild-type proteins, NiV-Ml-F and LayV-Cl-F, andone prefusion-stabilized NiV-F construct, NiVop08 (36), were consistent with our previous reports (19). With each peak in a DSF profile indicating a conformational event, for the unstabilized F ectodomains we observed one transition between 45-65°C and one or more transitions at higher temperature, up to over 90°C (Figure 4A). LayV-Cl-F showed two transitions with a negative peak at 50°C and a positive peak at 86°C. NiV-Ml-F showed three transitions with a negative peak 56°C, and positive peaks at 76°C, and 92°C. NiVop08 showed two transitions, a negative peak at 66°C and a positive peak at 76°C. We next measured the DSF profiles of the same set of proteins after pre-incubation at 60°C for 90 minutes, based on previous observations that heat treatment could force conformational conversion in Paramyxoviridae F proteins (40 conf). Overlaying all DSF profiles revealed that the lower-temperature negative peak present in all three proteins initially was eliminated in only the two wild-type proteins after heat incubation (Figure 4A). NSEM irmed that the wild type proteins have been fully converted to the postfusion conformation, as revealed by the characteristic elongated six-helix bundle present in the 2D class averages (Figure 4B). The NiVop08 prefusion stabilized construct retained the more compact shape characteristic of the prefusion state (Figure 4B). By exposing NiV-Ml-F and LayV-Cl-F to a series of times and temperatures, we found that 50°C, 15 minutes was the minimum amount of heat treatment sufficient to eliminate the pre-to-post- fusion conversion DSF peak, indicating that the F protein population was predominantly in the postfusion state. Together, these results indicate that the negative DSF peak present around 45-60°C is indicative of a pre- to postfusion conformational change taking place, and this DSF signature can be utilized to follow population level conformational 375 transition from the pre- to postfusion forms.

[0437] We then measured all purified F proteins for which there was sufficient material purified, typically under 5 pg, with DSF. We observed a wide range of conversion peak intensities and temperatures (Figures 4C-4F). While all proteins generated a large positive peak at high temperature indicative of a denaturation event, not all proteins produced a detectable conversion peak. Fourteen proteins had essentially no detectable signal prior to the high temperature peak (Figure 4D). For these F ectodomain samples, two-dimensional class averagesfrom NSEM analysis revealed the predominance of postfusion conformation for these proteins, with no detectable pre- fusion F species observed in the datasets, confirming the DSF results. Though not exclusively, most of the proteins with no conversion peak are Parahenipaviruses, and inversely, all of the Parahenipaviruses have no conversion peak, except for Langya virus strains. Many Parahenipaviruses F proteins could not be assessed due to insufficient purification yield, and, while not wishing to be bound by any theory, these results may indicate it could be due to inherent instability. Among the F proteins that generated a conversion peak, there were shifts in temperature and intensity (Figure 4E). These changes did not appear related to species differences, with NiV-Ml-F and NiV-M3-F, separated only by one amino acid mutation, being right- and left-shifted compared to the average, respectively. Both Langya virus strains were the most left-shifted of the set, indicative of lower pre-prefusion F stability. NSEM analysis showed a mix of pre- and postfusion conformations in this set, therefore the presence of this negative DSF peak indicates some amount of prefusion conformation present, rather than presence of a uniform set of prefusion proteins. Between the conversion peak and the final high temperature peak is typically a broad, positive peak, present with differing intensities and shapes. Further investigation would be required to determine if this peak is either part of any natural, yet undefined, conformational event or part of the denaturation process.

[0438] Two proteins of the panel, AngV-F and the sequence-corrected version of GhV-F that restores the signal peptide, GhV(+A)-F, display DSF profiles that are outliers to the rest of the panel (Figure 4F). Interestingly, both proteins displayed a profile that appeared closer to NiVop08, with their high temperature positive peak significantly left-shifted. Two-dimensional NSEM classes for these two proteins showed the presence of largely, but not exclusively, prefusion conformations. Despite their apparent similarities to a pre-fusion stabilized F construct, these are wild-type proteins that are can undergo normal conformational conversion, barring unexpected and dramatic mechanistic differences. The ability of the AngV-F and GhV(+A)-F proteins to undergo conformational conversion from the pre- to post-fusion states is supported by presence of post-fusion F forms in the NSEM 2D classes. We incubated these outlier F proteins under both the 60°C, 90 minute and 50°C, 15 minute conditions. After incubation at 50°C for 15minutes, a condition under which NiV-F and Lay V-F proteins show effectively complete transition to the post-fusion conformation, we found that AngV-F and GhV(+A)-F proteins retain much of their native DSF profile, suggesting greater pre-fusion stability. Incubation at 60°C for 90 minutes, however, was sufficient to induce a dramatic change in their DSF profiles, suggesting more complete conformational conversion.

[0439] Since DSF is reliant on primarily Tryptophan residues to generate signal, differences in DSF profile could arise from differing number and position of Tryptophans, however, the F ectodomain has only one Tryptophan that is almost universally conserved, with only CedV-l-F mutated at this spot. Furthermore, AngV-F contains one additional tryptophan unique to AngV- F. All F ectodomain constructs additionally contain one tryptophan in the foldon tag used to assist with trimerization and two in the twin-strep tag used for purification. In general, sequence variability is likely not a significant factor in DSF analysis of HNV-F proteins. Our results support the utilization of DSF as a rapid analytical technique requiring a small amount of protein for assessing population-level pre-to-post fusion transition of HNV-F proteins. In this study, the DSF analysis was central to our identification of two species that exhibit F pre-fusion conformations that appear more stabilized than in other HNV species.EXAMPLE 6

[0440] Cryo-EM Structures of AngV-F Protein

[0441] Based upon both amino acid sequence and pre-fusion stability as revealed by DSF analysis, AngV-F has divergent properties from the currently characterized F proteins. Therefore,, we determined the structure of the AngV-F protein by cryo-EM single particle analysis (Figure 5A). The cryo-EM dataset revealed three distinct particle populations. The most populated class was of the isolated F ectodomain trimer, followed by a dimer-of-trimers population, and finally a population where the F proteins formed a hexameric array. The isolated AngV-F trimer was refined to a resolution of 4.0 A and used for detailed examination of the F ectodomain structure and for comparing to the previously known structures of other F proteins. In cryo-EM structures of the HNV-F proteins the stalk region is typically poorly defined (15, 19,80, 71, 82). Our AngV-F cryo-EM structure revealed well-defined density for the stalk, allowing atomic-level modeling.

[0442] The AngV F ectodomain exhibited a larger central cavity compared to NiV-F and LayV-F proteins. This expanded cavity is the result of both the stalk region not inserting as far into the cavity as in other HNV-F proteins and the domains surrounding the cavity being spaced further apart (Figure 5B). In the previously determined HNV-F structures, the helical stalk region begins with a lysine at its N-terminal end, thus positioning a trio of lysines, one from each of the protomers, at the bottom of the internal cavity. In AngV-F, we instead observe a proline at the beginning of the stalk (HRB) helix (Figure 5C). The surface electrostatics of this region are therefore, different at this position for AngV-F compared to NiV-F and LayV-F structures. The proximity of the lysines in LayV-F and NiV-F and their electrostatic repulsion could be a source of metastability of the pre-fusion F protein, as charge repulsion may be a mechanism to force apart the HRB helix during conformational conversion. The absence of this lysine trio and the incorporation of a proline cap at the N terminus of the HRB helix may lend greater stability to the pre-fusion form of the AngV-F protein. Notably, all species in our panel have either a lysine or arginine at this site, with only AngV-F substituting the proline.

[0443] Several glycans could be resolved (N62, N93, N353, N434 and N458), including ones analogous to those previously defined in NiV-F structures (41, 42). The glycans shield known sites of vulnerability in Paramyxoviridae F proteins, including at the apex (N61), near the fusion peptide (N93 in Domain III), and on the stalk domain (N458). Interestingly, the glycan proximal to the fusion peptide utilizes a non-canonical NNV sequence (Figure 5D). The MojV-F and SHNV2-F proteins also have NNV sequences at this site that may be similarly glycosylated. Glycans in the apex region near N61 and the HRB / stalk region near N458 are observed in many HNV-F protein structures. However, there is no AngV-F analogue to the N414 glycan of HeV-F and NiV-F. Instead, AngV-F contains two new glycans in DI (N434) and DII (N353). The non- canonical NNV glycan at N93 is also found at the same site in HeV-F and NiV-F, but interestingly, not LayV-F. The differential glycosylation patterns observed across diverse HNV- F species may play a role in determining the potential for cross reactivity, but critically, thepresence of the NNV glycan in AngV-F indicates that prediction of glycosylation through sequence alone may be difficult.

[0444] Interactions between the AngV-F trimers were mediated through intermolecular contacts along the DI and Dill domains. These include hydrophobic interactions in Dill between L157 and V158 on neighboring ectodomains, both positioned on a loop previously identified as an important antigenic site (11, 38) (Figure 5E). Furthermore, there are charge interactions involving H45 from DI and T99 and Fl 05 of Dill (Figure 5E). Similar oligomeric states between pre-fusion F protein molecules (trimer, dimer-of-trimers, trimer-of-trimers) were also found in the cryo-EM dataset of a Hendra virus F protein (HeV-a2-F). AngV-F dimers resemble "mirror-like" protein dimers, with each exterior interaction surface of a protomer available for dimerization. These dimers, in turn, interact with protomers from three other trimers, forming hexameric structures. Each trimer is consistently aligned within the plane of the hexameric assembly, leading to a horizontally level lattice, distinct from an NiV-F crystal structure in which the lattice was not within one plane (41).

[0445] Taken together, our cryo-EM structures revealed that, while maintaining the overall conserved HNV-F architecture as observed in previously characterized HNV-F proteins, key mutations imparted notably distinct pre-fusion characteristics on the AngV-F protein. They also revealed multimerized pre-fusion F proteins adding to the growing body of literature that indicate a role for the multimeric assemblies in the context of the virion and indicate a distinct mode of F protein multimerization leading to the formation of hexameric lattices for the AngV-F protein.EXAMPLE 7

[0446] Structure-guided Pre-fusion Stabilization of diverse HNV-F Proteins

[0447] We investigated whether the proline substitution observed at the start of the HRB helix, unique to AngV-F, was the cause of the increased pre-fusion stability. We expressed a series of HNV-F constructs from a variety of HNV species incorporating this proline substitution. Available HNV-F structures indicate that the site analogous to K453 in NiV-Ml-F is consistently positioned at the N-terminal start of HRB, and even among other species forwhich structures are not available, the region surrounding this site is relatively well-conserved (Figure 6A).

[0448] All of the proline mutants were able to be purified in sufficient amounts to permit downstream quantitation, with yields per transfection volume mostly exceeding those of the wild-type construct (Figures 6B-D). In the case of the SHNV5-1-K455P mutant, the yield sharply decreased, but yet all mutants, including SHNV5-1-K445P-F, DSF analysis revealed significant pre-fusion stabilization (Figures 6B-D). The prominent fusion conversion peak was present in all constructs, often with greater intensity than the wild-type, or demonstrating a rightshift of conversion temperature, possibly indicating higher stability of the pre-fusion conformation. In the case of the NiV, HeV, and Lay V mutants, the 70-80°C peak was also present with higher intensity than the wild-type. Whereas GakV-l-F had displayed exclusively post-fusion conformation through both DSF and NSEM analysis (Figure 6D), the GakV-1- K452P-F DSF profile was dramatically altered, even appearing similar to the profile of the prefusion NiVop08 mutant or outliers like AngV-F and GhV(+A)-F. NSEM analysis confirmed the findings from DSF, with pre-fusion 2D classes observable with all mutants. The altered 2D classes of GakV-l -K452P-F, as with it’s DSF profile, was particularly striking. Whereas the wild-type classes contained solely post-fusion conformations, the mutant classes were almost universally pre-fusion. Despite the strong pre- fusion stabilization across all species, it was still common to observe post-fusion classes, reflecting that as with AngV-F, this mutation stabilizes pre-fusion conformation without preventing conversion (Figure 6B-D).

[0449] We assessed the binding of the NiV and HeV-based mutants, NiV-Ml-K458P-F and HeV- al.2-K453P-F, to the pre-fusion specific 4B8 antibody. Strong binding was observed between 4B8 IgG and all F constructs tested: both wild-type and mutant NiV-Ml-F and HeV- al.2-F, and NiVop08. However, after incubation at 50°C for 15 minutes, while binding to NiVop08 was maintained, binding to wild-type NiV / HeV was nearly eliminated. The HeV proline mutatnt’ s binding to 4B8 was similarly eliminated, while the NiV mutant’s binding was only moderately reduced. Taken together, our results indicate that pre- fusion stabilization of NiV / HeV through the substitution for the HRB start proline lends greater pre-fusion statestability but still allows for conformational conversion and antigenicity consistent with wild-type constructs.

[0450] We collected larger NSEM datasets and reconstructed 3D classes to quantify pre- and post-fusion ratios for these mutants and their wild-type counterparts. In all cases, the proportion of pre-fusion particles was increased in the mutant construct ccompared to wild-type. Interestingly, even for GakV-l-F and SHNV5-1-F, species where no pre-fusion conversion peak was present in DSF, a small amount of pre-fusion classes were detected in the larger NSEM datasets. Most striking was the conversion of GakV-l-F from -10% pre-fusion classes, the least of any species, to over 75% 543 pre-fusion with the proline mutant, the most of any species. Although the use of NSEM results in low-resolution 3D volumes, it is still apparent that the typical pre- and post-fusion class shapes are observed in all species, indicating that the proline mutation has not significantly altered the pre-fusion conformation.

[0451] In summary, the introduction of this proline presents an effective and translateable means of pre-fusion stabilization for HNV-F proteins, and one that is expected to have minimal impact on any of the antigenic sites on the surface of the protein.EXAMPLE 8

[0452] Purification and Characterization of Diverse Henipavirus G Head Domains

[0453] To analyze our panel of HNV-G proteins, we focused on the isolated head domain, which is responsible for receptor binding and is an important antigenic site (11). To allow for the previously described head domain sequences to be secreted in a mammalian expression system, we added an artificial secretion signal (43), and appended a C-terminal 8x His tag. The G head domains were transiently transfected and affinity purified with cobalt resin affinity chromatography, followed by SEC. The typical SEC profile for head domain proteins reflects the simple nature of their single-domain fold. All proteins eluted with a consistent single main peak, typically with a small, higher molecular shoulder or peak that is co-eluted aggregates. Yields for the head domains ranged from 2.5 to 35.3 mg / L, with the majority above 10 mg / L.

[0454] It has previously been noted that Langya and Mojiang virus attachment proteins do not bind to Ephrin B2 or B3 ligands (EFNB2 / B3), the receptors used by Henipaviruses (20, 82). Through BLI and SPR experiments, we assayed the binding of all G head domains in our panel to recombinant Fc-tagged Ephrin-B2 and -B3. As expected, all Nipah and Hendra virus head domains bound to both B2 and B3, with stronger binding observed for B2 rather than B3 (Figure 7A). CedV-l-G and GhV-G also bound to B2, with very faint binding to B3. Consistent with the previous observations, Langya and Mojiang virus G head domains, as well as the rest of the panel, including all Parahenipaviruses, did not show any appreciable binding to either B2 or B3.

[0455] We assessed the antigenicity of our G head domain panel with a series of previously characterized anti-NiV / HeV-G antibodies: HENV-26 and -32 (16), ml02.3 and .4 (84), nAH1.3 (47), and 1E5 (48). We also included the same Griffithsin construct used with the F panel. The known anti-NiV / HeV antibodies demonstrated broad, strong binding to all tested NiV and HeV strains (Figure 7B). There were lower levels of binding observed for some of these antibodies outside of NiV / HeV strains, particularly with Ghana virus. Griffithsin showed moderate binding to several strains, albeit inconsistent and not seemingly linked to sequence similarity. It bound most tightly to the SHNV3-G and SHNV1 1-G head domains. Although it did not show consistent binding to NiV / HeV proteins, as with HNV-F, nevertheless, the ability to bind to several Parahenipavirus G proteins is notable, given the lack of known binders for these novel strains. Slight variations in binding were observed through ELISA, mostly in the form of low to moderate levels of 1E5 binding to GhV-G, DarV-K-G, and MelV-G 586.

[0456] We used DSF to assess the thermostability of the G head domains. Unlike F proteins, the head domains are not known to undergo a dramatic conformational change when heated, and most proteins generated a profile with a single peak between 60-70°C, with some small shifts in the inflection temperature that was largely consistent within species groupings. Several species generated profiles distinct from the most common result, such as broad, negative, or double peaks. Unlike the relatively high conservation of Tryptophan positions in F proteins, G proteins display a much higher level of sequence diversity. Indeed, there are substantial differences in Tryptophan positions in species that generated outlier profiles, CedV, GhV, AngV, and SHNV2especially. The variability of Tryptophans in the G protein across species groupings precludes genus-wide comparisons of DSF profiles, yet these results provide interesting comparative insights within species groupings that may have implications for differences in receptor binding and virus fitness between species. As observed in the case of the SARS-CoV-2 spike protein, where thermostability of the receptor binding domain (RBD) is a property that is optimized during virus evolution (44-46), stability of the G head domain may impact its ability to bind receptor and antibodies, thus impacting its biological function.EXAMPLE 9

[0457] Structure of Gamak virus attachment protein head domain

[0458] HNV-G proteins are tetramers (dimer of dimers), with each monomer, in the ectodomain, composed of a helical stalk domain, a neck domain involved in oligomerization and control of quaternary structure, and a 6-bladed, P-propeller head domain where the receptorbinding site is located (11).

[0459] When comparing the sequence diversity of the HNV-G proteins, we noted that the C-terminal end of G head domains are a site of substantial variability (Figure 7C). GakV-G has among the longest C-terminal end, over 30 residues longer than NiV-G. A crystal structure of the GakV-2-G protein head domain at 1.4 A resolution (Figure 7D) showed the expected 6-bladed P-propeller architecture that is well-conserved among HNV-G proteins. A well-resolved glycan was observed in the region that is analogous to the Ephrin (EFN) receptor binding surface in the NiV-G head domain (Figure 7E). The presence of this glycan further illustrates the changed receptor specificity of the Parahenipaviruses, as it would likely cause a steric hindrance to Ephrin binding if present in NiV or HeV. The elongated C-terminal end was resolved in our structure, revealing a novel domain spanning residues 606 to 628 that formed a three-stranded P-sheet stacked against blade 5 of the P-propeller (Figure 7D). This additional compact and ordered domain is distinct from the previously known structures of Henipavirus G protein head domains. Furthermore, the N-terminus of the head domain appeared well ordered and packed against the 6-blade P-propeller core (Figure 7G). Residues 194-199 threaded between blades 5 and 6,contributing a P-strand to blade 6 as it continued to lead into blade 1. This stretch of residues also contacted blades 2, 4, and 5, thus forming a connection between different secondary structures within the -propeller. Taken together, the structure of the GakV G head domain illustrates the structural diversity that can be accommodated within the conserved - propeller fold to possibly impact receptor tropism and antigenicity.

[0460] Over the past few years, there has been a rapid expansion of identified species and strains in the Henipavirus genus, ultimately leading to the separation of these species into the Henipavirus and Parahenipavirus genera (3). Given the history of zoonotic transmission with these viruses, charcterization of these new species is an important first step for pandemic readiness. The present invention discloses a diversified panel of HNV F and G proteins that reflects the global diversity of HNVs. Until now, many of the strains in our expanded panel of Henipaviruses were known by their sequences alone, their surface glycoproteins totally uncharacterized and their relationships to other Henipaviruses unknown. Focusing particularly on Parahenipavirsues, closely related to Henipaviruses such as NiV and HeV, we elucidate their genetic diversity, antigenic relationships, structural novelties and mechanistic features.

[0461] Purifying and characterizing the F protein ectodomains of these strains has revealed that despite low sequence identity, HNV-F proteins share many architectural and biophysical similarities, a trend we previously noted by comparing Langya virus to Nipah virus (19). One of the notable similarities was the presence of some level of oligomerization between F trimers. This was apparent from the SEC profiles, but the presence of trimer multimers did not seem to correlate with other parameters measured, such as purification yield, metastability, or phylogeny. It appears instead that the formation of these complexes is transient and dependent on the concentration and environment of the F protein, as indicated by the absence of very small proportions of oligomers detected by mass photometry or NSEM where sample concentration is much lower than during size-exclusion chromatography and cryo-EM. The ubiquity of this type of interaction indicates that oligomerization of F proteins likely is a conserved trait within HNVs.

[0462] F protein oligomerization was pronounced in the case of Angavokely virus, where the formation of a hexameric lattice of F protein trimers was observed in the cryo-EM datasets. The existence of a hexamer-of-trimers arrangment has been noted previously with NiV (41), though the biological relevance was limited by the use of crosslinking to stabilize the arrangement. Our structure demonstrates the ability of AngV-F to multimerize in this way without any such assistance and, critically, without the involvement of the transmembrane domain, viral envelope, cytoplasmic domain, or viral matrix protein. The hexameric lattice is formed purely through ectodomain interactions, validating the characterizations of inter-trimer interactions previously reported for NiV (39), yet distinct in the implicated residues, with the AngV-F interaction involving DI and Dill, while the NiV-F interaction is dominated by DII residues. Previous studies have defined the lattice of Paramyxoviridae and Pneumoviridae F proteins, with various arrangements being determined. In one case, hexameric lattices have been observed, as with a whole-virion tomography structure of parainfluenza virus (3, 40). In others, such as with measles virus and respiratory syncytial virus, alternate arrangements of fusion and attachment proteins have been observed (52,53). Our results with AngV-F indicate that HNV-F proteins may adopt the hexameric arrangement, though structural studies of pseudoviruses and whole virions would help elucidate the physiological relevance of this finding. Furthermore, it cannot be ruled out that the formation of a hexameric F lattice may comprise one stage of HNV- F functionality, as the arrangment could shift during some type of maturation event or during G mediated triggering. The inventive characterizations of HNV-F proteins gives us a rapid and inexpensive tool with which we can assess their metastability without relying on time-consuming structure determination.

[0463] Using this assay, we were able to identify distinct signatures for pre-to-post fusion conversion and F protein denaturation, revealing how differences in sequence or structure could impact metastability. Though there are limitations in biological relevance working with soluble F0 ectodomains, this assay has proven reliable in quickly detecting altered pre-fusion stability in HNV-F proteins. It was through this DSF assay that we identified the strongly pre-fusion stabilized character of AngV-F, leading to our discovery of the HRB helix proline substitution asa translatable means for pre-fusion stabilization. Previous attempts to translate successful prefusion stabilization strategies for NiV-F to Parahenipaviruses have required structure-based evaluation of analogous sites to the disulfide, cavity-filling, or proline substitution strategies used previously (20,39). However, the region surrounding the start of the HRB helix as well as specifically the positively charged residue substituted for a proline are so well-conserved as to have allowed successful and efficacious transfer of the stabilizing mutant to each species attempted without further decision on positioning of the substitution. Additionally, while strategies like the triple mutation found in NiVop08 appear to fully lock the F protein in a prefusion conformation, the HRB start proline more moderately stabilizes without completely preventing conformational conversion. Though the incorporation of such a mutation would impart significant limitations on the biological relevance of any studies that evaluate conformational change in a mutated F construct, its utility is highlighted in situations where wild-type constructs do not produce any pre-fusion protein whatsoever, as we have seen with SHNV5-1-F and GakV-l-F.

[0464] DSF analysis of G head domains also reveals a diversity of themostability patterns. Although G head domains are comprised of a single domain 6-stranded P-propeller fold and are not known to have any major internal conformational rearrangements as part of their receptor binding mechanism, some studies have indicated that receptor binding by HNV-G proteins could effect subtle changes in protein flexibility that do not manifest as large-scale conformational events (54). The differences in thermostability could potentially correlate with mechanistic differences between strains; however, more structural data detailing the structural changes that take place after receptor binding in G proteins will be needed to confirm this. From our crystal structure of the Gamak virus head domain, we can observe other likely sources of mechanistic differences between species.

[0465] Though receptors for the Parhanehipaviruses remain unkown, the presence of the glycan in the traditional Ephrin-binding suggests an altered mode of receptor engagement. While the glycan may not be solely responsible for the lack of Ephrin binding in GakV-G, it would likely hinder binding of a similar protein receptor in the EFNB2 / 3 site, suggesting that GakVeither uses a smaller receptor or perhaps a different receptor binding site entirely. This would not be without precedent in Paramyxoviruses. For example, though EFNB2 / 3 engages the same face of the head domain in NiV and HeV that sialic acid does in other Paramyxoviruses (49,50), the measles virus receptor, CD 150, binds at the side surface of the head domain, making major contacts along blades 5 and 6 (51).

[0466] The C-terminal diversity in the head domains, and especially the additional exterior blade revealed in the GakV-2-G structure, may be the source of mechanistic changes as well. Together with the altered arrangement of the N- and C-terminal ends of the head domain we observed, these features could suggest a different pattern for quaternary interactions in GakV-G ectodomain tetramers, different types of conformational changes within GakV-G after receptor binding, or may even serve as a possible binding site for the as of yet undiscovered receptor.

[0467] While several anti-NiV and anti-HeV neutralizing antibodies have been well characterized, the antigenicity of new HNV strains had not yet been explored. By assessing the binding of a panel of both previously identified anti-HNV antibodies and newly discovered antibodies elicited through vaccination, we can now better describe the antigenic landscape of HNV proteins. As expected, we saw little ability of anti-NiV / HeV antibodies to bind to non- NiV / HeV proteins.

[0468] We found antigenic overlap between the F proteins of LayV, MojV, SHNV5, and to a lesser extent, GakV. Both a previously reported anti-Moj V-F antibody, 4G5, and our vaccine- elicited antibodies, 22F5 especially, bound to these strains. When considering our phylogenetic analysis, both by full genome and amino acid sequence, it can be seen this cross-reactivity spans distantly related strains. SHNV5 appears to be more

[0469] While the G and F proteins are not particularly heavily glycosylated relative to other viral glycoproteins, they ultimately do exhibit a varied glycosylation landscape, with glycans appearing in important antigenic sites in some species. While the lectin griffithsin only showed moderate levels of binding to most antigens tested here, it was able to bind to a very diverse set of HNV-G head domains. Most interestingly, its strongest binding was to the SHNV3 and SHNV1 1-G head domains, proteins for which there are currently no known antibodies or ligandsthat can bind them. As investigation of griffithsin for antiviral potency across several families continues, it may prove a useful tool for analysis of novel HNV proteins.

[0470] With a group of viruses prone to such sequence diversity, one major concern in the event of a possible widespread outbreak of disease in humans is that emergent infective strains may escape immunity conferred from prior infection or immunization with other HNV strains. This phenomenon can be observed with yearly outbreaks of new influenza strains or with the progression of SARS-CoV-2 evolution, where new strains emerged and quickly dominated the pool of circulating virus as they evaded prior immunity (85,86). A focus of efforts to address this possibility have long been designing immunogens that can elicit antibodies with broad coverage of the currently exisitng and, hopefully, future diversity of strains. Being able to draw antigenic boundaries within the Henipavirus and Parahenipavirus genera is a foundation for similar efforts, and here we see that while the prospect of cross-reactivity over the entire genus remains elusive, there is already demonstrated cross-reactivity among a large portion of the Parahenipaviruses. While it will be important to assess this observed cross-reactivity in neutralization assays and animal challenge models, there is currently a lack of knowledge both of the host receptors for Parahenipaviruses and of the proper cell lines or animal models to study infection.

[0471] While some progress has been made in identifying permissive cell lines for Parahenipaviruses (83,87), the rate of fusion is typically much lower than seen with more well- studied Nipah / Hendra models, indicating that such viral entry assays could be in need of further optimization. Ultimately, in the absence of robust experimental systems to study the virological properties and the breadth of protection from potential HNV infections, the inventions reported here build a critical foundation for future exploration of Parahenipavirus antigenicity.

[0472] Ultimately, this broad characterization of HNV proteins provides a set of valuable biophysical and biochemical data for many species and strains that, to this point, had only been known as a sequence in a database. As these new species are studied in more depth, we hope that our analysis can be used to provide context for the comparison of other key aspects of Henipavirus biochemistry, structure, and immunology.

[0286] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein.

[0287] The use of the terms “a” and “an” and “the” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any nonclaimed element as essential to the practice of the invention.

[0288] Preferred embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.References1. Rima, B. et al. ICTV Virus Taxonomy Profile: Paramyxoviridae. J Gen Virol 100, 1593-1594 (2019). doi.org / 10.1099 / jgv.0.0013282. 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Claims

Claims:

1. A modified Henipavirus fusion (F) protein having a proline at an amino acid position analogous to position 447 of the Angavokely virus (AngV), optionally wherein the modified Henipavirus F protein is selected from the group consisting of SEQ ID NOs: 70-193.

2. A method for identification of Henipavirus fusion (F) protein antigens comprising: a) preparing an identification and classification scheme for Henipavirus strains; b) aligning the amino acid sequences of F proteins, wherein the amino acid sequences of the F proteins were truncated at the C-terminal of the ectodomain equivalent to Langya virus residue 447; c) using the alignments of b) to create a phylogenic tree to identify viral strains specific to fruit bat reservoirs and shrew reservoirs; d) preparing a panel of F proteins by preparing DNA expression vectors encoding ectodomain residues spanning residues 1 to the equivalent of 488 in Nipah virus; and e) isolating and purifying the F proteins expressed by the constructs.

3. The method of claim 2, wherein the F proteins of the expression constructs of (e) are tested for antigenicity against known antibodies.

4. The method of claim 2, wherein the F proteins of the expression constructs of (e) are used to immunize mice to create antibodies.

5. The method of claim 2, wherein the F proteins of the expression constructs of (e) are characterized biophy sically.

6. The method of either claim 3 or 4, wherein the F proteins of the expression constructs are modified to comprise the amino acid sequence of SEQ ID NOS: 70-131, 132-193, or a portion or fragment thereof.

7. A method for identification of Henipavirus G protein antigens comprising: a) preparing an identification and classification scheme for Henipavirus strains; b) aligning the amino acid sequences of G proteins, wherein the N-terminal portion of the ectodomains, up to the end of the transmembrane domain,was deleted, and wherein the amino acid sequence of G head domains analogous to Nipah virus residue 71 was the starting point; c) using the alignments of b) to create a phylogenic tree to identify viral strains specific to fruit bat reservoirs and shrew reservoirs; d) preparing a panel of G proteins by preparing DNA expression vectors encoding the isolated head domain; and e) isolating and purifying the G proteins expressed by the constructs.

8. The method of claim 7, wherein the G proteins of the expression constructs of (e) are tested for antigenicity against known antibodies.

9. The method of claim 7, wherein the G proteins of the expression constructs of (e) are used to immunize mice to create antibodies.

10. The method of claim 7, wherein the G proteins of the expression constructs of (e) are characterized biophysically.

11. An immunogenic composition, comprising the Henipavirus fusion protein antigen of the expression constructs of (e) of claim 1 and a pharmaceutically acceptable carrier, optionally wherein the F proteins have been modified to have a proline at a position analogous to position 447 in AngV.

12. The immunogenic composition of claim 11, wherein the composition comprises the amino acid sequence of SEQ ID NOS: 70-131, 132-193, or a portion or fragment thereof.

13. An immunogenic composition, comprising the Henipavirus G protein antigen of the expression constructions of (e) of claim 7 and a pharmaceutically acceptable carrier.

14. A method for generating an immune response in a subject, comprising administering to the subject an effective amount of the immunogenic composition of claim 12.

15. An amino acid sequence encoding the Henipavirus fusion protein antigen of the expression constructs of (e) of claim 2, optionally wherein the F proteins have been modified to have a proline at a position analogous to position 447 in AngV16. The amino acid sequence of claim 15, wherein the sequence is selected from the group consisting of SEQ ID NOS: 70-131 and 132-193.

17. A nucleotide sequence encoding the amino acid sequence encoding the Henipavirus fusion protein antigen of the expression constructs of (e) of claim 118. A nucleotide sequence encoding the amino acid sequence of the Henipavirus fusion protein antigen, wherein the amino acid sequence is selected from the group consisting of SEQ ID NOS: 70-131 and 132-19319. A vector encoding the nucleotide sequence of either of claim 17 or 18.

20. A cell comprising the sequence of either claim 17 or 18.

21. An immunogenic composition comprising a full-length Henipavirus F protein having a proline amino acid inserted at the HRB helix start analogous to the proline in the AngV-F wild type F protein (position 447 in AngV).

22. The immunogenic composition of claim 21, comprising the amino acid sequences of SEQ ID NOS: 70-131.

23. The immunogenic composition of claim 21, comprising the amino acid sequences of one or more of SEQ ID NOS: 70-131 which have been modified from the wild-type protein by insertion of the proline amino acid or substitution of an amino acid for the proline amino acid at the position equivalent to position 447 in AngV.

24. An immunogenic composition comprising a Henipavirus F ectodomain protein truncation having a proline amino acid inserted at the HRB helix start analogous to the proline in the AngV-F wild type F protein (position 447 in AngV).

25. The immunogenic composition of claim 24, comprising one or more of the amino acid sequences of SEQ ID NOS: 132-193.

26. The immunogenic composition of claim 24, comprising one or more of the amino acid sequences of SEQ ID NOS: 132-193 which have been modified from the wild-type by insertion of the proline amino acid or substitution of an amino acid for the proline amino acid at the position equivalent to position 447 in AngV.

27. An antibody, comprising 22F5 or 1C8.

28. The antibody 22F5 of claim 27, wherein said antibody having a VH domain comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 68 and a VL domain comprising the amino acid sequence having at least 90% identity to SEQ ID NO: 69.

29. A composition comprising antibody 22F5 and a pharmaceutically acceptable carrier.

30. The use of the composition of claim 29 to identify LayV-F ectodomains.

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