A SARS-CoV-2 pseudovirus

Abstract

The present application relates to the field of genetic engineering and molecular biology, in particular to a truncated Spike protein or a variant thereof, a fusion protein comprising the protein, a vector, a host cell and a virus-like particle. The virus-like particle can be used for screening of antibodies and drugs against SARS-CoV-2 (e.g. detecting the neutralization activity of SARS-CoV-2 antibodies, or screening drugs that inhibit SARS-CoV-2 infection at the cellular level). The present application also relates to a pharmaceutical composition and a kit comprising the above-mentioned protein.

Description

Technical Field

[0001] This invention relates to the fields of genetic engineering and molecular biology, specifically to truncated Spike proteins or variants thereof, fusion proteins comprising said proteins, and virus-like particles. The virus-like particles can be used for screening antibodies and drugs against SARS-CoV-2 (e.g., detecting the neutralizing activity of SARS-CoV-2 antibodies, or screening drugs that inhibit SARS-CoV-2 infection at the cellular level). This invention also relates to methods and kits for preparing the aforementioned proteins. Background Technology

[0002] Pseudoviruses are infectious virus-like particles that resemble real viruses, formed by the encapsulation of heterologous nucleic acids carrying reporter genes by viral capsid or envelope proteins. Because the encapsulated nucleic acid does not have the complete nucleic acid sequence to replicate and form a virus, pseudoviruses only have one round of infectivity, making them relatively safe to handle.

[0003] Pseudoviruses are primarily formed by co-transfection of 293T cells with multiple plasmids. They replicate, express, and assemble within the cells, and are obtained by collecting culture supernatant or lysing cells. Pseudoviruses not only possess the same infectivity as true viruses and enter cells in the same way, but they also lack the ability to replicate and produce viruses after entering the cell. Therefore, they are harmless and can be used more safely for the detection of neutralizing antibodies and antiviral drugs.

[0004] For highly infectious and pathogenic viruses, such as SARS-CoV-2, Ebola virus, rabies virus, and H7N9, research on real viruses is extremely dangerous and the quantity is limited. Pseudoviruses, on the other hand, are safe and can be mass-produced through transfection technology, which facilitates research on these highly dangerous viruses.

[0005] The characteristics of pseudovirus systems are that they overcome the shortcomings of traditional methods, making detection technology safer, simpler, faster, and with higher throughput. Moreover, they enable the detection of some viruses that are difficult to culture.

[0006] However, when viral membrane proteins are used directly for pseudovirus construction, the viral titer is often low, which limits the application of pseudoviruses. Therefore, it is necessary to further improve the viral titer of pseudoviruses. Summary of the Invention

[0007] Through extensive experimentation and repeated exploration, the inventors of this application truncated and modified the Spike protein, thereby obtaining a high-titer SARS-CoV-2 pseudovirus. Based on this, they further modified the vector through numerous experiments, obtaining a modified vector and transfection conditions for preparing the high-titer pseudovirus, thus completing this invention.

[0008] Therefore, in a first aspect, the present invention provides a truncated Spike protein or a variant thereof, wherein, compared with the Spike protein of wild SARS-CoV-2, (1) the C-terminus of the truncated Spike protein is shortened by 13-23 amino acids, and / or, (2) the amino acids of the truncated Spike protein are missing at the positions corresponding to positions 669-672 of SEQ ID NO:1;

[0009] The variant, compared to the truncated Spike protein, has one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids; e.g., conserved substitutions of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids).

[0010] In some implementations, the truncated Spike protein has 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, and 23 amino acids truncated at its C-terminus compared to the wild-type SARS-CoV-2 Spike protein.

[0011] In some embodiments, the truncated Spike protein, compared to the wild-type SARS-CoV-2 Spike protein, also contains a deletion, namely, the amino acid at the positions corresponding to positions 669-672 of SEQ ID NO:1 is missing.

[0012] In some embodiments, the missing amino acid is a furin restriction site.

[0013] In some embodiments, the missing amino acid is PRRA.

[0014] In some embodiments, compared to the wild-type SARS-CoV-2 Spike protein, the truncated Spike protein has amino acids missing at positions corresponding to positions 669-672 of SEQ ID NO:1.

[0015] In some implementations, the truncated Spike protein has 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, and 23 amino acids truncated at its C-terminus compared to the wild-type SARS-CoV-2 Spike protein.

[0016] In some embodiments, the missing amino acid is a furin restriction site.

[0017] In some embodiments, the missing amino acid is PRRA.

[0018] In some embodiments, the truncated Spike protein or a variant thereof has an amino acid sequence selected from the following:

[0019] (1) SEQ ID NO:2-SEQ ID NO:23;

[0020] (2) An amino acid sequence having one or more amino acid substitutions, deletions, or additions (e.g., substitutions, deletions, or additions of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids; e.g., conservative substitutions of 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10 amino acids) compared to any of the sequences shown in SEQ ID NO:2-SEQ ID NO:23;

[0021] (3) An amino acid sequence having at least 95%, at least 96%, at least 97%, at least 98%, or at least 99% sequence identity with any of the sequences shown in SEQ ID NO:2-SEQ ID NO:23.

[0022] In this invention, the aforementioned variants essentially retain the biological functions of the proteins from which they originate. In this invention, the biological functions primarily refer to the biological functions of the wild-type SARS-CoV-2 Spike protein, including but not limited to, its binding activity to the cell surface receptor (ACE2), its ability to undergo conformational changes (e.g., coiling and folding to form hairpin-like structures), and its self-assembly into virus-like particles.

[0023] In a second aspect, the present invention provides a fusion protein comprising a truncated Spike protein or a variant thereof as described above, and additional polypeptides.

[0024] In some embodiments, the additional polypeptide is optionally linked to the N-terminus or C-terminus of the protein via a linker. Such linkers are well known in the art, and examples include, but are not limited to, peptide linkers comprising one or more (e.g., 1, 2, 3, 4, or 5) amino acids (e.g., Glu or Ser). In some embodiments, the peptide linker is flexible. In some embodiments, a flexible peptide linker may be advantageous, as it can link two protein / peptide components while maintaining their respective activity and function. Such peptide linkers include, but are not limited to, (GGGGS)3.

[0025] In some embodiments, the additional polypeptide is selected from tags, signal peptides or guide peptides, detectable markers (e.g., luciferase (fluc), green fluorescent protein (GFP)), or any combination thereof. In some embodiments, the protein of the present invention or a variant thereof may be linked to a tag sequence to facilitate the expression, detection, and / or purification of the protein of the present invention. In some embodiments, the protein of the present invention or a variant thereof may be linked to a signal peptide or guide peptide sequence to guide the secretion of the protein of the present invention. In some embodiments, the protein of the present invention or a variant thereof may be linked to a detectable marker sequence to facilitate the detection or tracing of the protein of the present invention.

[0026] In some embodiments, the signal peptide is selected from natural signal peptides of Spike protein, CD5, CD14, IgG heavy chain, immunoglobulin light chain, tissue plasminogen activator (tPA), SCGB1D1, serum albumin preproprotein, Glucose, IL-2, and IFNα2.

[0027] In some embodiments, the fusion protein of the present invention includes a tag known to those skilled in the art, including but not limited to histidine tags (His), glutathione thiotransferase tags (GST), influenza virus hemagglutinin (HA), and flag-tags. Those skilled in the art know how to select an appropriate tag based on the desired purpose (e.g., protein expression, purification, or detection).

[0028] In some embodiments, the signal peptide has an amino acid sequence selected from the following: SEQ ID NO:24-SEQ ID NO:33.

[0029] In some embodiments, the tag has an amino acid sequence as shown in SEQ ID NO:35.

[0030] The aforementioned fusion protein / other polypeptide (e.g., tag or signal peptide) will not adversely affect the desired activity of the protein of the present invention (e.g., the activity of binding to the cell surface receptor (ACE2), the ability to undergo conformational changes (e.g., curling and folding to form a hairpin-like structure), or self-assembly into a virus-like particle).

[0031] In a third aspect, the present invention provides an isolated nucleic acid molecule that encodes a truncated Spike protein or a variant thereof as described above, or that encodes a fusion protein as described above.

[0032] In a fourth aspect, the present invention provides a vector comprising the nucleic acid molecules described above. The vector of the present invention can be a cloning vector or an expression vector. In some embodiments, the vector of the present invention is, for example, a plasmid, a granulocyte, a bacteriophage, a Cosmid, etc. In some embodiments, the vector is capable of expressing the protein of the present invention or the fusion protein described above within a host cell (e.g., a mammalian cell, such as a human cell).

[0033] In some embodiments, the vector is a eukaryotic expression plasmid containing the nucleic acid molecules described above.

[0034] In some embodiments, the eukaryotic expression plasmid is selected from pcDNA3.1+, pCAG3.1, and pCMV3.1.

[0035] In a fifth aspect, the present invention provides a host cell comprising the nucleic acid molecule as described above or the carrier as described above.

[0036] In some embodiments, the cells are mammalian cells.

[0037] In some embodiments, the mammalian cells are human cells, such as hematopoietic cells, epithelial cells, hepatocytes, tumor cells, and nerve cells.

[0038] In some embodiments, the cells are cells that express or overexpress human ACE2 protein (e.g., 293T-ACE2).

[0039] In some embodiments, the host cell also includes a packaging plasmid.

[0040] In a sixth aspect, the present invention provides a virus-like particle containing a truncated Spike protein or a variant thereof as described above, or a fusion protein as described above; or, the virus-like particle is composed of a truncated Spike protein or a variant thereof as described above, or a fusion protein as described above.

[0041] In this invention, the truncated Spike protein or its variants, or the fusion protein, self-assembles to form the outer shell of the virus-like particle. In some embodiments, the virus-like particle further contains nucleic acid components. Such nucleic acid components are, for example, encapsulated within the outer shell formed by the self-assembly of the truncated Spike protein or its variants, or the fusion protein. In some embodiments, the nucleic acid components are selected from any one or more of the following:

[0042] (a) Exogenous DNA, or exogenous DNA encoding exogenous proteins or polypeptides, or exogenous DNA encoding RNA;

[0043] (b) A vector that optionally comprises the exogenous DNA described in (a);

[0044] (c) Exogenous RNA, or exogenous RNA encoding exogenous proteins or polypeptides;

[0045] (d) A vector that optionally contains the exogenous RNA described in (c).

[0046] In this invention, the nucleic acid component can be double-stranded, single-stranded, or any combination thereof. In this invention, the nucleic acid component can be DNA, RNA, or any combination thereof.

[0047] In some embodiments, the nucleic acid component is co-transfected into a host cell with the vector described above and undergoes self-assembly, where the protein expressed by the host cell encapsulates the nucleic acid to form virus-like particles.

[0048] In some implementations, the nucleic acid component is a packaging plasmid.

[0049] In a seventh aspect, the present invention provides a method for preparing a SARS-CoV-2 pseudovirus, the method comprising expressing a truncated Spike protein or a variant thereof, or a fusion protein as described above, in a suitable host cell. In some embodiments, a vector as described above is transferred into a suitable host cell, and the truncated Spike protein or a variant thereof, or a fusion protein as described above, is expressed. In some embodiments, the vector as described above and a packaging plasmid are co-transferred into the host cell.

[0050] In some implementations, the vector and packaging plasmid described above are transfected into host cells.

[0051] In some embodiments, the mass ratio of the vector used in the transfection process to the packaging plasmid is 1-3:1-4, for example, 3:1, 2:1, 1:1, 1:2, 1:3, or 1:4. In some embodiments, the mass ratio of the vector used in the transfection process to the packaging plasmid is 1:3.

[0052] In some implementations, the transfection reagent used in the transfection process is selected from: lipofectamine 2000, lipofectamine 3000, PEI, or any combination thereof.

[0053] In some embodiments, the host cell is a human cell, such as a hematopoietic cell, epithelial cell, hepatocyte, tumor cell, or nerve cell.

[0054] In some embodiments, the cells infected by the virus-like particles are cells that overexpress human ACE2 protein (e.g., 293T-ACE2).

[0055] In an eighth aspect, the present invention provides a kit comprising one or more components selected from the following: a truncated Spike protein or a variant thereof as described above, a fusion protein as described above, an isolated nucleic acid molecule as described above, a vector as described above, a host cell as described above, and a virus-like particle as described above.

[0056] In some embodiments, the kit also includes a packaging plasmid.

[0057] In some embodiments, the kit also includes a transfection reagent.

[0058] In some embodiments, the transfection reagent is selected from: lipofectamine2000, lipofectamine3000, PEI, or any combination thereof.

[0059] In some embodiments, the kit of the present invention also includes instructions for use. Various optimization schemes are discussed herein to improve the titer of the prepared pseudoviruses. For example, the selection of the packaging plasmid, the ratio of the expression plasmid to the packaging plasmid, the selection of the transfection reagent, the ratio of the transfection reagent to the plasmid, and the pseudovirus harvesting time, etc. All of the above schemes can be included in the instructions for use of the kit of the present invention.

[0060] In a ninth aspect, the present invention provides a method for detecting the neutralizing activity of a SARS-CoV-2 antibody, the method comprising contacting the virus-like particle with the antibody or the host cell before, simultaneously with, or after contacting the virus-like particle as described above with the host cell.

[0061] Compared to direct ELISA and single-epitope competitive ELISA, the pseudovirus method better reflects the neutralizing ability of antibodies. It simulates the biological process by which antibodies block viral entry into cells, directly reflecting the protective effect of the antibody. This method typically involves contacting the host cell or virus-like particles with the antibody before the pseudovirus is introduced into the host cell (i.e., before infecting the host cell with the pseudovirus), and then detecting the ability of the pseudovirus to infect the host cell. If the pseudovirus's infectivity decreases, it indicates that the antibody inhibits pseudovirus infection of cells and provides protection to the cells.

[0062] In a tenth aspect, the present invention provides a method for screening candidate drugs capable of inhibiting SARS-CoV-2 infection of cells, the method comprising contacting the virus-like particles or host cells with the candidate drug before, simultaneously or after contacting the virus-like particles as described above with the host cells.

[0063] In some embodiments, the method typically involves contacting the candidate drug to the host cell or virus-like particles before the pseudovirus is exposed to the host cell (i.e., before infecting the host cell with the pseudovirus), and then detecting the ability of the pseudovirus to infect the host cell. A decrease in pseudovirus infectivity indicates that the candidate drug inhibits pseudovirus infection of the cell and provides protection to the cell.

[0064] In an eleventh aspect, the present invention provides the use of the truncated Spike protein or variant thereof as described above, the fusion protein as described above, the isolated nucleic acid molecule as described above, the vector as described above, the host cell as described above, and the virus-like particle as described above for detecting or screening anti-SARS-CoV-2 antibodies or drugs.

[0065] Terminology Definition

[0066] In this invention, unless otherwise stated, the scientific and technical terms used herein have the meanings commonly understood by those skilled in the art. Furthermore, the operational steps used herein, such as molecular genetics, nucleic acid chemistry, chemistry, molecular biology, biochemistry, cell culture, microbiology, cell biology, genomics, and recombinant DNA, are all conventional steps widely used in their respective fields. To better understand this invention, definitions and explanations of relevant terms are provided below.

[0067] As used herein, the term "SARS-CoV-2" is short for "Severe Acute Respiratory Syndrome Coronavirus 2," formerly known as "Novel Coronavirus" or "2019-nCov." It belongs to the β-coronavirus genus and is an enveloped, single-stranded, positive-sense RNA virus. The genome sequence of SARS-CoV-2 is known to those skilled in the art and can be found, for example, in GenBank: MN908947. SARS-CoV-2 contains at least three membrane proteins, including a surface spike protein (S), an integrated membrane protein (M), and a membrane protein (E). Like SARS-CoV, SARS-CoV-2's receptor specifically binds to angiotensin-converting enzyme 2 (ACE2) on host cells via the receptor-binding domain (RBD) on the S protein, enabling viral membrane fusion and cell entry. The S protein plays a crucial role in viral infection of cells.

[0068] As used in this article, the terms “surface spike protein,” “Spike protein,” and “S protein” refer to a membrane protein of SARS-CoV-2, and they have the same meaning and are used interchangeably.

[0069] As used in this article, the terms “novel coronavirus pneumonia” and “COVID-19” refer to pneumonia caused by SARS-CoV-2 infection. They have the same meaning and can be used interchangeably.

[0070] As used herein, the term "SARS-CoV" or "SARS-CoV-1" is short for "severe acute respiratory syndrome coronavirus 1," which belongs to the genus *β-coronavirus* and is an enveloped, single-stranded, positive-sense RNA virus. The genome sequence of SARS-CoV-1 is known to those skilled in the art and can be found, for example, in GenBank: AAP13567.1. Pneumonia caused by SARS-CoV-1 is called SARS.

[0071] As used herein, the terms “wild” and “natural” are used interchangeably. When these terms are used to describe nucleic acid molecules, peptides, or proteins, they mean that the nucleic acid molecule, peptide, or protein exists in nature, is found in nature, and has not undergone any artificial modification or processing. As used herein, wild-type SARS-CoV-2 Spike protein refers to a naturally occurring, biologically active Spike protein. Those skilled in the art can readily obtain the amino acid sequence of the Spike protein from various public databases, such as GenBank. For example, the amino acid sequence of wild-type SARS-CoV-2 Spike protein may be shown in SEQ ID NO: 1.

[0072] As used in this article, the term "C-terminal truncated X amino acids" means that the last X amino acids at the C-terminus are truncated.

[0073] As used herein, the term "corresponding position" refers to the amino acid position at the equivalent position in two sequences being compared when performing an optimal alignment, i.e., when aligning two sequences to obtain the highest percentage identity. For example, the expression "position corresponding to positions 669-672 of SEQ ID NO:1" means the amino acid position in the compared sequence that is equivalent to positions 669-672 of SEQ ID NO:1 when performing an optimal alignment of a sequence with SEQ ID NO:1, i.e., when aligning a sequence with SEQ ID NO:1 to obtain the highest percentage identity.

[0074] As used herein, the terms "pseudovirus" and "virus-like virus" have the same meaning and are used interchangeably; they refer to a virus-like particle formed by the self-assembly of viral proteins, which either does not encapsulate nucleic acid or encapsulates other nucleic acids. Therefore, while the pseudovirus or virus-like virus can infect host cells, it does not have the ability to replicate autonomously. Consequently, it has higher biosafety compared to a real virus. The packaging system of a pseudovirus generally consists of two parts: a packaging component and an expression component. The packaging component is constructed from the viral genome (e.g., HIV-1) with the genetic information required for packaging, reverse transcription, and integration removed, providing the proteins necessary for the pseudovirus particle. The expression component is complementary to the packaging component, containing the genetic information required for packaging, reverse transcription, and integration, as well as the exogenous target gene. By co-transfecting host cells with the packaging component and the vector component, pseudovirus particles can be harvested from the cell supernatant.

[0075] As used herein, the terms "backbone plasmid" and "packaging plasmid" have the same meaning and are used interchangeably. As is generally understood by those skilled in the art, a viral vector system (particularly a lentiviral vector system) can consist of two parts: a packaging component (e.g., a packaging plasmid or backbone plasmid) and a vector component (e.g., a recombinant expression vector carrying a target gene); wherein the packaging component (e.g., a packaging plasmid or backbone plasmid) provides all the accessory proteins required for transcription and packaging of genetic material (e.g., RNA) into recombinant pseudoviral particles. Thus, high-titer pseudoviral particles can be generated by co-transfecting cells with a recombinant expression vector and a packaging plasmid, followed by pseudoviral packaging within the cells, and then secretion of the packaged pseudoviral particles into an extracellular culture medium. Such packaging plasmids or backbone plasmids are well known to those skilled in the art. For example, backbone plasmids constructed based on HIV-1 include, but are not limited to, pSG3.Δenv (Wei X et al., Antibody neutralization and escape by HIV-1. Nature 422:307-312, 2003) and NL4-3.Fluc.R-.E- (Connor RI et al., Vpr is required for efficient replication of human immunodeficiency virus type-1 in mononuclear phagocytes. Virology 206:935–944, 1995), as well as pSG3.Δenv.Fluc and pSG3.Δenv.cmvFluc constructed in the embodiments of this application.

[0076] As used herein, the term "identity" refers to the sequence matching between two polypeptides or two nucleic acids. Two compared sequences are identical at a position when the same base or amino acid monomeric subunit occupies the same location (e.g., a position in each of two DNA molecules is occupied by adenine, or a position in each of two polypeptides is occupied by lysine). The "percentage identity" between two sequences is a function of the number of matching positions shared by the two sequences divided by the number of positions compared × 100. For example, if six out of ten positions in two sequences match, then the two sequences have 60% identity. For example, the DNA sequences CTGACT and CAGGTT share 50% identity (three out of six positions match). Typically, two sequences are compared to produce the maximum identity. Such comparisons can be made using methods readily available, for example, computer programs such as the Align program (DNAstar, Inc.) Needleman et al. (1970) J. Mol. Biol. 48: 443-453. The percentage identity between two amino acid sequences can also be determined using the algorithm of E. Meyers and W. Miller (Comput. Appl Biosci., 4:11-17 (1988)) integrated into the ALIGN program (version 2.0), which uses a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Alternatively, the percentage identity between two amino acid sequences can be determined using the Needleman and Wunsch algorithm (J MoIBiol. 48:444-453 (1970)) in the GAP program integrated into the GCG software package (available at www.gcg.com), which uses a Blossum 62 matrix or a PAM250 matrix, along with gap weights of 16, 14, 12, 10, 8, 6, or 4, and length weights of 1, 2, 3, 4, 5, or 6.

[0077] As used herein, the term "conservative substitution" means an amino acid substitution that does not adversely affect or alter the biological activity of a protein / peptide containing an amino acid sequence. For example, conservative substitutions can be introduced using standard techniques known in the art, such as site-directed mutagenesis and PCR-mediated mutagenesis. Conservative amino acid substitutions include substitutions of amino acid residues with amino acid residues having similar side chains, such as substitutions with residues that are physically or functionally similar to the corresponding amino acid residues (e.g., having similar size, shape, charge, chemical properties, including the ability to form covalent or hydrogen bonds). Families of amino acid residues with similar side chains have been defined in the art. These families include amino acids with basic side chains (e.g., lysine, arginine, and histidine), acidic side chains (e.g., aspartic acid and glutamic acid), uncharged polar side chains (e.g., glycine, asparagine, glutamine, serine, threonine, tyrosine, cysteine, and tryptophan), nonpolar side chains (e.g., alanine, valine, leucine, isoleucine, proline, phenylalanine, and methionine), β-branched side chains (e.g., threonine, valine, and isoleucine), and aromatic side chains (e.g., tyrosine, phenylalanine, tryptophan, and histidine). Therefore, it is preferable to replace the corresponding amino acid residue with another amino acid residue from the same side chain family. Methods for identifying conserved amino acid substitutions are well known in the art (see, for example, Brummell et al., Biochem. 32:1180-1187 (1993); Kobayashi et al., Protein Eng. 12(10):879-884 (1999); and Burks et al., Proc. Natl Acad. Set USA 94:412-417 (1997), which are incorporated herein by reference).

[0078] As used herein, the term "vector" refers to a nucleic acid delivery vehicle into which polynucleotides can be inserted. When a vector enables the expression of a protein encoded by the inserted polynucleotide, it is called an expression vector. Vectors can be introduced into host cells through transformation, transduction, or transfection, allowing the genetic material elements they carry to be expressed in the host cells. Vectors are well-known to those skilled in the art and include, but are not limited to: plasmids; phage particles; Cos plasmids; artificial chromosomes, such as yeast artificial chromosomes (YAC), bacterial artificial chromosomes (BAC), or P1-derived artificial chromosomes (PAC); bacteriophages such as λ phage or M13 phage; and animal viruses. Animal viruses that can be used as vectors include, but are not limited to, retrotranscriptoviruses (including lentiviruses), adenoviruses, adeno-associated viruses, herpesviruses (such as herpes simplex virus), poxviruses, baculoviruses, papillomaviruses, and papillomaviruses (such as SV40). A vector may contain multiple elements controlling expression, including but not limited to, promoter sequences, transcription initiation sequences, enhancer sequences, selection elements, and reporter genes. Additionally, a vector may contain a replication initiation site.

[0079] As used herein, the term "host cell" refers to a cell that can be used to introduce a vector, including but not limited to prokaryotic cells such as Escherichia coli or Bacillus subtilis, fungal cells such as yeast cells or Aspergillus, insect cells such as S2 Drosophila cells or Sf9, or animal cells such as fibroblasts, CHO cells, COS cells, NSO cells, HeLa cells, BHK cells, HEK 293 cells, or human cells.

[0080] Those skilled in the art will understand that the design of expression vectors can depend on factors such as the choice of host cells to be transformed and the desired expression level. A vector can be introduced into a host cell to produce transcripts, proteins, or peptides, including proteins, fusion proteins, isolated nucleic acid molecules, etc., as described herein.

[0081] As used herein, the term "effective amount" means an amount that is effective in achieving the intended purpose. For example, an effective amount for the prevention or treatment of a disease (e.g., rotavirus infection) means an amount that is effective in preventing, stopping, or delaying the onset of a disease (e.g., rotavirus infection), or in alleviating, reducing, or treating the severity of an existing disease (e.g., a disease caused by rotavirus infection). Determining such an effective amount is within the capabilities of those skilled in the art. For example, an effective amount for therapeutic purposes will depend on the severity of the disease to be treated, the overall state of the patient's own immune system, the patient's general characteristics such as age, weight, and sex, the method of administration of the drug, and other concurrent treatments, etc.

[0082] As used herein, the term "neutralizing activity" refers to the functional activity of an antibody or antibody fragment to bind to antigenic proteins on a virus, thereby preventing the virus from infecting cells and / or maturing and / or releasing viral progeny. Antibodies or antibody fragments with neutralizing activity can prevent viral amplification, thereby inhibiting or eliminating viral infection.

[0083] Beneficial effects of the invention

[0084] Compared with existing technologies, the pseudovirus titer of SARS-CoV-2 prepared in this application is increased by at least 10 times, with a maximum increase of 826 times. The development of this pseudovirus expands the scope of SARS-CoV-2 research beyond laboratories with high biosafety conditions. Furthermore, the pseudovirus can be used to detect antibody neutralization capacity, providing excellent technical support for the evaluation of SARS-CoV-2 vaccines and drugs, as well as research on the viral pathogenesis mechanism.

[0085] The embodiments of the present invention will now be described in detail with reference to the accompanying drawings and examples. However, those skilled in the art will understand that the following drawings and examples are for illustrative purposes only and are not intended to limit the scope of the invention. Various objects and advantages of the present invention will become apparent to those skilled in the art from the following detailed description of the drawings and preferred embodiments. Attached Figure Description

[0086] Figure 1 The fluorescence values ​​(RLU) of pseudoviruses prepared from the 22 truncated Spike proteins in Example 1 are shown.

[0087] Figure 2 The fluorescence value (RLU) of the pseudovirus prepared by fusing spike protein with 10 signal peptides is shown.

[0088] Figure 3 The fluorescence values ​​(RLU) of pseudoviruses prepared from the three modified vectors are shown.

[0089] Figure 4 The fluorescence values ​​(RLU) of pseudoviruses prepared from the four modified vectors are shown.

[0090] Figure 5 The fluorescence values ​​(RLU) of pseudoviruses prepared from three backbone plasmids are shown.

[0091] Figure 6 The fluorescence values ​​(RLU) of pseudoviruses prepared using three different transfection reagents are shown.

[0092] Figure 7The fluorescence values ​​(RLU) of pseudoviruses prepared with different mass ratios of recombinant vector to backbone plasmid are shown.

[0093] Sequence information

[0094] Information on some of the sequences involved in this invention is provided in Table 1 below.

[0095] Table 1: Sequence Description

[0096]

[0097]

[0098]

[0099]

[0100]

[0101]

[0102]

[0103]

[0104]

[0105]

[0106]

[0107]

[0108] Detailed Implementation

[0109] The invention will now be described with reference to the following embodiments, which are intended to illustrate the invention (and not limit it).

[0110] Unless otherwise specified, the experiments and methods described in the embodiments are performed in accordance with conventional methods well known in the art and described in various references. For example, conventional techniques such as immunology, biochemistry, chemistry, molecular biology, microbiology, cell biology, genomics, and recombinant DNA used in this invention can be found in Sambrook, Fritsch, and Maniatis, *Molecular Cloning: A Laboratory Manual*, 2nd edition (1989); *Current Protocols in Molecular Biology* (edited by FM. Ausubel et al., (1987)); the *Methods in Enzymology* series (academic publishing company): *PCR 2: A PRACTICAL APPROACH* (edited by MJ. MacPherson, BD. Hames, and GR. Taylor, (1995)); and *Animal Cell Culture*. CELLCULTURE (edited by R.R. Freshney (1987)).

[0111] Furthermore, unless specific conditions are specified in the examples, conventional conditions or conditions recommended by the manufacturer should be followed. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products. Those skilled in the art will understand that the examples are described by way of illustration and are not intended to limit the scope of protection claimed by the invention. All disclosures and other references mentioned herein are incorporated herein by reference in their entirety.

[0112] Example 1. Construction of the recombinant vector pcDNA3.1-Spike

[0113] Based on the BamH1 and Xho1 restriction sites on the plasmid pcDNA3.1+ (purchased from Invitrogen, catalog number V79020), the nucleotide sequence encoding the SARS-CoV-2 Spike protein (SEQ ID NO:1) and the nucleotide sequence encoding the natural signal peptide (SEQ ID NO:34) were ligated to the pcDNA3.1+ plasmid to construct the recombinant vector pcDNA3.1-Spike (Genebank: MT613044).

[0114] The specific procedure is as follows: The double-digested plasmid pcDNA3.1+ was recovered, mixed with the target fragment, infused with infusion enzyme, and placed on a PCR instrument at 50°C for 15 minutes for ligation. The ligation product was then transferred into 50 μl of DH5α competent cells and incubated on ice for 30 minutes. After removing the competent cells, they were heat-shocked in a 42°C water bath for 45-60 seconds and then immediately placed on ice for 2 minutes. Add 500 μl of antibiotic-free LB medium and incubate at 37°C with shaking at 120 rpm for 1 hour. Centrifuge the competent cells at 2800 rpm for 2 min, discard 500 μl of supernatant, mix the remaining cells and spread them on ampicillin-resistant (Amp) plates. Incubate overnight (14-16 hours) at 37°C with the plates inverted. Pick single colonies and inoculate them into 5 mL of LB medium (100 μg / mL Amp). Incubate at 37°C with shaking at 220 rpm for 14-16 hours. Store in glycerol and send to a sequencing company for identification of the expression plasmid.

[0115] Example 2. Modification and titer test of Spike protein

[0116] 1. Modification of Spike protein

[0117] A key difference between the SARS-CoV-2 Spike protein and the SARS-CoV-1 Spike protein lies in the addition of four amino acids, PRRA, at the S1 and S2 cleavage sites. This addition forms a new furinase cleavage site, corresponding to amino acids 669-672 of SEQ ID NO:1. Therefore, this embodiment modifies the SARS-CoV-2 Spike protein and constructs a recombinant vector containing the modified Spike protein. The modification of the Spike protein is as follows:

[0118] (1) The intracellular region of the C-terminus of Spike protein was shortened by 13, 14, 15, 16, 17, 18, 19, 20, 21, 22 and 23 amino acids, respectively. The amino acid sequence corresponding to the shortened Spike protein is shown in SEQ ID NO:2-12. Recombinant vectors were constructed according to the steps described in Example 1 and named pcDNA3.1-Spike13 to pcDNA3.1-Spike23, respectively.

[0119] (2) The Furin enzyme cleavage site of the Spike protein was deleted, that is, the amino acid corresponding to position 669-672 of SEQ ID NO:1 was deleted (i.e., PRRA). The amino acid sequence corresponding to the modified Spike protein is shown in SEQ ID NO:13. The recombinant vector constructed accordingly was named pcDNA3.1-furin.

[0120] (3) First, the C-terminal intracellular region of the Spike protein was truncated by 13, 14, 15, 16, 17, 19, 20, 21, 22, and 23 amino acids, respectively. Then, the furinase restriction site was deleted, specifically the amino acid 669-672 of SEQ ID NO:1 (i.e., PRRA). The amino acid sequence of the modified Spike protein is shown in SEQ ID NO:14-23. The corresponding recombinant vectors were named pcDNA3.1-Spike13-furin to pcDNA3.1-Spike23-furin, respectively.

[0121] The specific process is as follows: Based on the principle of circular PCR, the sequence of the vector pcDNA3.1-Spike (Genebank: MT613044) was used as a template for PCR sequence amplification. Since the PCR product contains the template strand, to reduce interference and improve the success rate of subsequent experiments, the template strand was excised using the Dpn I restriction endonuclease and incubated overnight at 37°C (the principle is that the template DNA sequence originates from conventional E. coli and has undergone dam methylation modification, making it sensitive to Dpn I endonuclease and thus fragmented; while the DNA amplified in vitro has not undergone methylation and therefore will not be recognized by Dpn I). The PCR product was constructed into the pcDNA3.1+ plasmid according to the method described in Example 1 and transformed into DH5α competent cells, plated, and cultured. Approximately 5 single colonies were picked from each culture plate and transferred to 5 mL of LB liquid medium containing ampicillin, and cultured at 220 rpm and 37°C for approximately 16 hours. Plasmids were extracted in small quantities from the bacterial culture, and the extracted plasmids were subjected to agarose gel electrophoresis for preliminary identification based on band size. Bands with correct preliminary identification results were sent for sequencing to further determine whether the mutation was successful and to proceed with the subsequent pseudovirus construction.

[0122] 2. Titer test

[0123] According to the instructions for Lipofectamine 3000 transfection reagent (Thermo Fisher Scientific, cat.no. L3000015), the same amount of the 22 recombinant vectors constructed above (with vector pcDNA3.1-Spike as a control) and the backbone plasmid pSG3.Δenv.cmvFluc (this plasmid was constructed in our laboratory and contains a luciferase gene (fluc) carrying a CMV promoter; related information and sequence of this plasmid are disclosed in patent CN201510293955.5) were co-transfected into 293T cells. The supernatant was collected after approximately 48 hours, and the collected pseudoviruses were used to infect 293T-hACE2 cells (purchased from Sinocare). After 48 hours, the supernatant was discarded, and the chemiluminescence value (RLU) was detected and calculated using the Britelite plus reporter geneassay system (PerkinElmer, cat.no. 6066769) luciferase detection kit.

[0124] The results are as follows Figure 1 As shown, truncating the intracellular region of the Spike protein by 13-23 amino acids increased the viral titer by approximately 10-fold. Deleting the furin restriction site increased the viral titer by approximately 40-fold. Deleting both simultaneously resulted in a further increase in viral titer. Specifically, simultaneously deleting the furin restriction site and truncating 21 amino acids from the intracellular region of the Spike protein's C-terminus resulted in the highest chemiluminescence value (RLU), calculated to be 826-fold higher than the control.

[0125] Example 3. Modification of the recombinant vector and titer test

[0126] 1. Replacement of signal peptide

[0127] To investigate the effect of different signal peptide substitutions on pseudovirus titers, ten signal peptides (i.e., the signal peptides of CD5, CD14, IgG heavy chain, Immunoglobulin light chain, tPA, SCGB1D1, Serum albumin preproprotein, Gluc, IL-2, and IFNα2) were used to replace the SARS-CoV-2 Spike protein's own signal peptide (SEQ ID NO: 34). The amino acid sequences corresponding to the ten signal peptides are shown in SEQ ID NO: 24-33, respectively. Following the experimental procedures described above, modified vectors were constructed, and pseudoviruses were packaged into these vectors according to the aforementioned steps. Viral titers were then tested.

[0128] The results are as follows Figure 2As shown, compared with the control vector pcDNA3.1-Spike (2019-nCoV), the pseudoviruses constructed from vectors containing the signal peptide CD5 and Immunoglobulin light chain showed significantly higher titers, with a 3-5 fold increase in titer.

[0129] 2. Expression vehicle

[0130] To investigate the effect of different expression vectors on viral titer, this embodiment constructed four different vectors based on three plasmids. Following the experimental steps described above, the constructed vectors were encapsulated as pseudoviruses to test viral titer. The plasmids used are as follows: (1) pCAG3.1 plasmid, the specific information of which is disclosed in patent CN201510293955.5; (2) pCMV3.1 plasmid, the specific information of which is disclosed in patent CN201510293955.5; (3) pcDNA3.1+. The following vectors were constructed by inserting the nucleotide sequence encoding the Spike protein (SEQ ID NO:1) and the nucleotide sequence encoding its signal peptide (SEQ ID NO:34) into the multiple cloning site of the above plasmid: (1) pCAG3.1-Sipke vector, which expresses the Sipke protein as shown in SEQ ID NO:1 and the signal peptide as shown in SEQ ID NO:34; (2) pCAG-Sipke-tag vector, which expresses the Sipke protein as shown in SEQ ID NO:1 and the signal peptide as shown in SEQ ID NO:34, and in addition, a tag as shown in SEQ ID NO:35 is expressed at the C-terminus of the Sipke protein; (3) pCMV3.1-Spike vector, which expresses the Sipke protein as shown in SEQ ID NO:1 and the signal peptide as shown in SEQ ID NO:34; (4) pcDNA3.1-Spike vector, which expresses the Sipke protein as shown in SEQ ID NO:1 and the signal peptide as shown in SEQ ID NO:34.

[0131] The results are as follows Figure 3 As shown, the pseudovirus prepared using the pCAG-Sipke-tag vector exhibits a significantly increased viral titer.

[0132] 3. Carrier Modification

[0133] Based on the above experimental results, the vector was further modified using three plasmids. The plasmids used were as follows: (1) pCAG3.1 plasmid; (2) pCMV3.1 plasmid; (3) pcDNA3.1+. Sipke protein or a modified Sipke protein was inserted into the multiple cloning sites of the above plasmids to construct the following vectors: (1) pCAG-Sipke-tag vector, which expresses the Sipke protein as shown in SEQ ID NO:1 and the signal peptide as shown in SEQ ID NO:34, and also expresses the tag as shown in SEQ ID NO:35 at the C-terminus of the Sipke protein; (2) pCAG-LC-furin-tag vector, which expresses the signal peptide as shown in SEQ ID NO:27, the Spike protein with the furin site deleted as shown in SEQ ID NO:13, and the tag as shown in SEQ ID NO:35; (3) pCAG-CD5-furin-tag vector, which expresses the signal peptide as shown in SEQ ID NO:24, the Spike protein with the furin site deleted as shown in SEQ ID NO:13, and the tag as shown in SEQ ID NO:35; (4) pCAG-furin vector, which expresses the Spike protein with the furin site deleted as shown in SEQ ID NO:13 and the tag as shown in SEQ ID NO:35. The signal peptide shown in NO:34; (5) pcDNA3.1-Spike vector (i.e. Figure 4 (The 2019-S vector in the middle).

[0134] The results are as follows Figure 4 As shown, the pCAG-LC-furin-tag vector can package pseudoviruses with higher titers, with a titer increase of 30-50 times compared to the control (2019-S vector).

[0135] Example 4. Exploration of pseudovirus envelope conditions

[0136] 1. Replacement of backbone plasmids

[0137] Following the steps outlined above, the recombinant vector pcDNA3.1-Spike and three backbone plasmids were co-transfected into 293T cells according to the instructions of the Lipofectamine 3000 transfection reagent. The supernatant was collected approximately 48 hours later, and the collected pseudoviruses were used to infect 293T-hACE2 cells. After 48 hours, the supernatant was discarded, and the chemiluminescence values ​​were detected using the Britelite plus reportergene assay system (PerkinElmer, cat.no.6066769) luciferase assay kit. The National Institutes of Health (NIH) provided two HIV-1-based backbone plasmids: pSG3.Δenv (Wei X, Decker JM, Wang S, Hui H, Kappes JC, Wu X, Salazar-Gonzalez JF, Salazar MG, Kilby JM, Saag MS, Komarova NL, Nowak MA, Hahn BH, Kwong PD and Shaw GM. Antibody neutralization and escape by HIV-1. Nature 422:307-312, 2003.) and NL4-3.Fluc.R-.E- (Connor RI, Chen BK, Choe S, Landau NR. Vpr is required for efficient replication of human immunodeficiency virus type-1 in mononuclear phagocytes. Virology 206:935–944, 1995). Based on this, we amplified the Fluc gene and constructed the pSG3.Δenv.Fluc vector carrying the Fluc gene. Then, we added the CMV promoter to the upstream of the Fluc gene to the pSG3.Δenv.Fluc vector to construct the pSG3.Δenv.cmvFluc vector (specific information about pSG3.Δenv.Fluc and pSG3.Δenv.cmvFluc is disclosed in patent CN201510293955.5). Therefore, the three backbone plasmids used in this embodiment are: (1) pNL4-3.Fluc.R-.E-; (2) pSG3.Δenv.Fluc; (3) pSG3.Δenv.cmvFluc.

[0138] The results are as follows Figure 5As shown, the pseudovirus titer prepared using the pSG3.Δenv.cmvFluc backbone plasmid was significantly higher than the others.

[0139] 2. Replacement of transfection reagent

[0140] Following the steps outlined above, the recombinant vector pcDNA3.1-Spike and the backbone plasmid pSG3.Δenv.cmvFluc were co-transfected into 293T cells using three different transfection reagents. The supernatant was collected approximately 48 hours later, and the collected pseudoviruses were used to infect 293T-hACE2 cells. After 48 hours, the supernatant was discarded, and the chemiluminescence values ​​were detected using a Britelite plus reporter gene assay system (PerkinElmer, cat.no.6066769) luciferase assay kit. The three transfection reagents used were lipofectamine 2000, lipofectamine 3000, and PEI.

[0141] The results are as follows Figure 6 As shown, the pseudoviruses prepared using the lipofectamine 3000 transfection reagent have higher titers.

[0142] 3. Ratio of recombinant vector to backbone plasmid

[0143] Following the steps outlined above, the recombinant vector pcDNA3.1-Spike and the backbone plasmid pSG3.Δenv.cmvFluc were co-transfected into 293T cells at different mass ratios according to the instructions of the Lipofectamine 3000 transfection reagent. The supernatant was collected approximately 48 hours later, and the collected pseudoviruses were used to infect 293T-hACE2 cells. After 48 hours, the supernatant was discarded, and the chemiluminescence values ​​were detected using the Briteliteplus reporter gene assay system (PerkinElmer, cat.no.6066769) luciferase assay kit. The mass ratios of the recombinant vector pcDNA3.1-Spike and the backbone plasmid pSG3.Δenv.cmvFluc were 3:1, 2:1, 1:1, 1:2, 1:3, and 1:4, respectively.

[0144] The results are as follows Figure 7 As shown, when the mass ratio of recombinant vector pcDNA3.1-Spike to backbone plasmid pSG3.Δenv.cmvFluc is 1:3, the prepared pseudovirus titer is higher.

[0145] Although specific embodiments of the invention have been described in detail, those skilled in the art will understand that various modifications and variations can be made to the details based on all the published teachings, and all such changes are within the scope of protection of the invention. The entire scope of the invention is given by the appended claims and any equivalents thereof. sequence list <110> China National Institutes for Food and Drug Control <120> A pseudovirus of SARS-CoV-2 <130> IDC200214 <160> 35 <170> PatentIn version 3.5 <210> 1 <211> 1261 <212> PRT <213> artificial <220> <223> SARS-CoV-2 Spike protein <400> 1 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp 1235 1240 1245 Ser Glu Pro Val Leu Lys Gly Val Lys Leu His Tyr Thr 1250 1255 1260 <210> 2 <211> 1248 <212> PRT <213> artificial <220> <223> Spike protein with 13 amino acids truncated at the C-terminus <400> 2 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp 1235 1240 1245 <210> 3 <211> 1247 <212> PRT <213> artificial <220> <223> Spike protein with 14 amino acids truncated at the C-terminus <400> 3 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp 1235 1240 1245 <210> 4 <211> 1246 <212> PRT <213> artificial <220> <223> C-terminally truncated Spike protein with 15 amino acids removed <400> 4 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu 1235 1240 1245 <210> 5 <211> 1245 <212> PRT <213> artificial <220> <223> Spike protein with 16 amino acids truncated at the C-terminus <400> 5 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe Asp 1235 1240 1245 <210> 6 <211> 1244 <212> PRT <213> artificial <220> <223> Spike protein with 17 amino acids truncated at the C-terminus <400> 6 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys Phe 1235 1240 <210> 7 <211> 1243 <212> PRT <213> artificial <220> <223> Spike protein with 18 amino acids truncated at the C-terminus <400> 7 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys Lys 1235 1240 <210> 8 <211> 1242 <212> PRT <213> artificial <220> <223> Spike protein with 19 amino acids truncated at the C-terminus <400> 8 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys Cys 1235 1240 <210> 9 <211> 1241 <212> PRT <213> artificial <220> <223> Spike protein with 20 amino acids truncated at the C-terminus <400> 9 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser Cys 1235 1240 <210> 10 <211> 1240 <212> PRT <213]] artificial <220> <223> Spike protein with 21 amino acids truncated at the C-terminus <400> 10 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly Ser 1235 1240 <210> 11 <211> 1239 <212> PRT <213> artificial <220> <223> Spike protein with 22 amino acids truncated at the C-terminus <400> 11 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys Gly 1235 <210> 12 <211> 1238 <212> PRT <213> artificial <220> <223> Spike protein with 23 amino acids truncated at the C-terminus <400> 12 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala 660 665 670 Arg Ser Val Ala Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly 675 680 685 Ala Glu Asn Ser Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr 690 695 700 Asn Phe Thr Ile Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr 705 710 715 720 Lys Thr Ser Val Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu 725 730 735 Cys Ser Asn Leu Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn 740 745 750 Arg Ala Leu Thr Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu 755 760 765 Val Phe Ala Gln Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp 770 775 780 Phe Gly Gly Phe Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro 785 790 795 800 Ser Lys Arg Ser Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu 805 810 815 Ala Asp Ala Gly Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile 820 825 830 Ala Ala Arg Asp Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val 835 840 845 Leu Pro Pro Leu Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala 850 855 860 Leu Leu Ala Gly Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala 865 870 875 880 Ala Leu Gln Ile Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly 885 890 895 Ile Gly Val Thr Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala 900 905 910 Asn Gln Phe Asn Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser 915 920 925 Thr Ala Ser Ala Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala 930 935 940 Gln Ala Leu Asn Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala 945 950 955 960 Ile Ser Ser Val Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu 965 970 975 Ala Glu Val Gln Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu 980 985 990 Gln Thr Tyr Val Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala 995 1000 1005 Ser Ala Asn Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly 1010 1015 1020 Gln Ser Lys Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met 1025 1030 1035 Ser Phe Pro Gln Ser Ala Pro His Gly Val Val Phe Leu His Val 1040 1045 1050 Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala 1055 1060 1065 Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe 1070 1075 1080 Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr 1085 1090 1095 Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn 1100 1105 1110 Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro 1115 1120 1125 Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr 1130 1135 1140 Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser 1145 1150 1155 Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg 1160 1165 1170 Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu 1175 1180 1185 Gln Glu Leu Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr 1190 1195 1200 Ile Trp Leu Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val 1205 1210 1215 Thr Ile Met Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys 1220 1225 1230 Gly Cys Cys Ser Cys 1235 <210> 13 <211> 1257 <212> PRT <213> artificial <220> <223> Spike protein with furin site deleted <400> 13 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro Val 1235 1240 1245 Leu Lys Gly Val Lys Leu His Tyr Thr 1250 1255 <210> 14 <211> 1244 <212> PRT <213> artificial <220> <223> Spike protein with 13 amino acids truncated at the C-terminus and the furin site deleted. <400> 14 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp 1235 1240 <210> 15 <211> 1243 <212> PRT <213> artificial <220> <223> Spike protein with 14 amino acids truncated at the C-terminus and the furin site deleted. <400> 15 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp 1235 1240 <210> 16 <211> 1242 <212> PRT <213> artificial <220> <223> Spike protein with 15 amino acids truncated at the C-terminus and the furin site deleted. <400> 16 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys Cys Lys Phe Asp Glu 1235 1240 <210> 17 <211> 1241 <212> PRT <213> artificial <220> <223> Spike protein with 16 amino acids truncated at the C-terminus and the furin site deleted. <400> 17 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys Cys Lys Phe Asp 1235 1240 <210> 18 <211> 1240 <212> PRT <213> artificial <220> <223> Spike protein with 17 amino acids truncated at the C-terminus and the furin site deleted. <400> 18 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys Cys Lys Phe 1235 1240 <210> 19 <211> 1238 <212> PRT <213> artificial <220> <223> Spike protein with 19 amino acids truncated at the C-terminus and the furin site deleted. <400> 19 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys Cys 1235 <210> 20 <211> 1237 <212> PRT <213> artificial <220> <223> Spike protein with 20 amino acids truncated at the C-terminus and the furin site deleted. <400> 20 Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser Cys 1235 <210> twenty one <211> 1236 <212> PRT <213> artificial <220> <223> Spike protein with 21 amino acids truncated at the C-terminus and the furin site deleted. <400> twenty one Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly Ser 1235 <210> twenty two <211> 1235 <212> PRT <213> artificial <220> <223> Spike protein with 22 amino acids truncated at the C-terminus and the furin site deleted. <400> twenty two Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys Gly 1235 <210> twenty three <211> 1234 <212> PRT <213> artificial <220> <223> Spike protein with 23 amino acids truncated at the C-terminus and the furin site deleted. <400> twenty three Ser Gln Cys Val Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr 1 5 10 15 Thr Asn Ser Phe Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg 20 25 30 Ser Ser Val Leu His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser 35 40 45 Asn Val Thr Trp Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr 50 55 60 Lys Arg Phe Asp Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe 65 70 75 80 Ala Ser Thr Glu Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr 85 90 95 Thr Leu Asp Ser Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr 100 105 110 Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe 115 120 125 Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu 130 135 140 Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser 145 150 155 160 Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn 165 170 175 Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr 180 185 190 Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe 195 200 205 Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr 210 215 220 Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly 225 230 235 240 Asp Ser Ser Ser Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly 245 250 255 Tyr Leu Gln Pro Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr 260 265 270 Ile Thr Asp Ala Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys 275 280 285 Cys Thr Leu Lys Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser 290 295 300 Asn Phe Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile 305 310 315 320 Thr Asn Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala 325 330 335 Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp 340 345 350 Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr 355 360 365 Gly Val Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr 370 375 380 Ala Asp Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro 385 390 395 400 Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp 405 410 415 Phe Thr Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys 420 425 430 Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn 435 440 445 Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly 450 455 460 Ser Thr Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu 465 470 475 480 Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr 485 490 495 Arg Val Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val 500 505 510 Cys Gly Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn 515 520 525 Phe Asn Phe Asn Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn 530 535 540 Lys Lys Phe Leu Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr 545 550 555 560 Thr Asp Ala Val Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr 565 570 575 Pro Cys Ser Phe Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr 580 585 590 Ser Asn Gln Val Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val 595 600 605 Pro Val Ala Ile His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr 610 615 620 Ser Thr Gly Ser Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly 625 630 635 640 Ala Glu His Val Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala 645 650 655 Gly Ile Cys Ala Ser Tyr Gln Thr Gln Thr Asn Ser Arg Ser Val Ala 660 665 670 Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser 675 680 685 Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile 690 695 700 Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val 705 710 715 720 Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu 725 730 735 Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr 740 745 750 Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln 755 760 765 Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe 770 775 780 Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser 785 790 795 800 Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly 805 810 815 Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp 820 825 830 Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu 835 840 845 Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly 850 855 860 Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile 865 870 875 880 Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr 885 890 895 Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn 900 905 910 Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala 915 920 925 Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn 930 935 940 Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val 945 950 955 960 Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln 965 970 975 Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val 980 985 990 Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn Leu 995 1000 1005 Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys Arg 1010 1015 1020 Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro Gln 1025 1030 1035 Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro 1040 1045 1050 Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp 1055 1060 1065 Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly 1070 1075 1080 Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile 1085 1090 1095 Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val 1100 1105 1110 Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu 1115 1120 1125 Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His 1130 1135 1140 Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala 1145 1150 1155 Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val 1160 1165 1170 Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly 1175 1180 1185 Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu Gly 1190 1195 1200 Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met Leu 1205 1210 1215 Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys Ser 1220 1225 1230 Cys <210> 24 <211> 24 <212> PRT <213> artificial <220> <223> CD5 signal peptide <400> 24 Met Pro Met Gly Ser Leu Gln Pro Leu Ala Thr Leu Tyr Leu Leu Gly 1 5 10 15 Met Leu Val Ala Ser Val Leu Ala 20 <210> 25 <211> 20 <212> PRT <213> artificial <220> <223> CD14 signal peptide <400> 25 Met Glu Arg Ala Ser Cys Leu Leu Leu Leu Leu Leu Pro Leu Val His 1 5 10 15 Val Ser Ala Thr 20 <210> 26 <211> 19 <212> PRT <213> artificial <220> <223> IgG heavy chain signal peptide <400> 26 Met Asp Trp Thr Trp Arg Val Phe Cys Leu Leu Ala Val Thr Pro Gly 1 5 10 15 Ala His Pro <210> 27 <211> 19 <212> PRT <213> artificial <220> <223> Immunoglobulin light chain signal peptide <400> 27 Met Ala Trp Ser Pro Leu Phe Leu Thr Leu Ile Thr His Cys Ala Gly 1 5 10 15 Ser Trp Ala <210> 28 <211> twenty two <212> PRT <213> artificial <220> <223> tPA signal peptide <400> 28 Met Asp Ala Met Lys Arg Gly Leu Cys Cys Val Leu Leu Leu Cys Gly 1 5 10 15 Ala Val Phe Val Ser Pro 20 <210> 29 <211> twenty one <212> PRT <213> artificial <220> <223> SCGB1D1 signal peptide <400> 29 Met Arg Leu Ser Val Cys Leu Leu Leu Leu Thr Leu Ala Leu Cys Cys 1 5 10 15 Tyr Arg Ala Asn Ala 20 <210> 30 <211> 18 <212> PRT <213> artificial <220> <223> Serum albumin preproprotein signal peptide <400> 30 Met Lys Trp Val Thr Phe Ile Ser Leu Leu Phe Leu Phe Ser Ser Ala 1 5 10 15 Tyr Ser <210> 31 <211> 17 <212> PRT <213> artificial <220> <223> Gluc's signal peptide <400> 31 Met Gly Val Lys Val Leu Phe Ala Leu Ile Cys Ile Ala Val Ala Glu 1 5 10 15 Ala <210> 32 <211> 20 <212> PRT <213> artificial <220> <223> IL-2 signal peptide <400> 32 Met Tyr Arg Met Gln Leu Leu Ser Cys Ile Ala Leu Ser Leu Ala Leu 1 5 10 15 Val Thr Asn Ser 20 <210> 33 <211> twenty three <212> PRT <213> artificial <220> <223> IFNα2 signal peptide <400> 33 Met Ala Leu Thr Phe Ala Leu Leu Val Ala Leu Leu Val Leu Ser Cys 1 5 10 15 Lys Ser Ser Cys Ser Val Gly 20 <210> 34 <211> 12 <212> PRT <213> artificial <220> <223> Spike protein signal peptide <400> 34 Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser 1 5 10 <210> 35 <211> 11 <212> PRT <213> artificial <220> <223> Label <400> 35 Gly Gly Thr Glu Thr Ser Gln Val Ala Pro Ala 1 5 10

Claims

1. A truncated Spike protein, in which, Compared with the wild-type SARS-CoV-2 Spike protein, (1) the C-terminus of the truncated Spike protein is shortened by 21 amino acids, and (2) the amino acids in the truncated Spike protein are deleted at the following positions: positions corresponding to positions 669-672 of SEQ ID NO: 1; wherein the amino acid sequence of the wild-type SARS-CoV-2 Spike protein is as shown in SEQ ID NO:

1.

2. The truncated Spike protein of claim 1, wherein, The missing amino acid is a furin restriction site.

3. The truncated Spike protein of claim 1, wherein, The missing amino acid is PRRA.

4. The truncated Spike protein according to any one of claims 1-3, wherein, The amino acid sequence of the truncated Spike protein is shown in SEQ ID NO:

21.

5. A fusion protein comprising the truncated Spike protein of any one of claims 1-4 and an additional polypeptide; said additional polypeptide is selected from tags, signal peptides or lead peptides, detectable markers, or any combination thereof.

6. The fusion protein of claim 5, wherein, The additional polypeptide is attached to the N-terminus or C-terminus of the protein via a linker.

7. The fusion protein of claim 5, wherein, The signal peptide is selected from the natural signal peptides of Spike protein, CD5, CD14, IgG heavy chain, immunoglobulin light chain, tissue plasminogen activator, SCGB1D1, serum protein precursor, Gluc, IL-2 and IFNα2, wherein the sequence of the natural signal peptide of Gluc is shown in SEQ ID NO:

31.

8. The fusion protein of claim 5, wherein, The tags include histidine tags, glutathione thiotransferase tags, influenza virus hemagglutinin, and flag tags.

9. The fusion protein of claim 5, wherein, The signal peptide has an amino acid sequence selected from the following: SEQ ID NO: 24-SEQ ID NO:

33.

10. The fusion protein of claim 5, wherein, The tag has an amino acid sequence as shown in SEQ ID NO:

35.

11. An isolated nucleic acid molecule encoding a truncated Spike protein as described in any one of claims 1-4, or encoding a fusion protein as described in any one of claims 5-10.

12. A vector comprising the nucleic acid molecule of claim 11.

13. The carrier according to claim 12, wherein, The vector is a eukaryotic expression plasmid containing the nucleic acid molecule of claim 11.

14. The carrier according to claim 13, wherein, The eukaryotic expression plasmids were selected from pcDNA3.1+, pCAG3.1 and pCMV3.

1.

15. A host cell comprising the nucleic acid molecule of claim 11 or the vector of any one of claims 12-14.

16. The host cell of claim 15, wherein, The host cell is a mammalian cell.

17. The host cell of claim 16, wherein, The cells of the mammal are human cells.

18. The host cell of claim 17, wherein, The cells in the human body are epithelial cells, liver cells, or tumor cells.

19. The host cell of claim 15, wherein, The host cell is a cell that expresses or overexpresses human ACE2 protein.

20. The host cell of claim 15, wherein, The host cell is a 293T cell that expresses or overexpresses human ACE2 protein.

21. The host cell of claim 15, wherein, The host cell also includes packaging plasmids.

22. A virus-like particle, said virus-like particle containing a truncated Spike protein as described in any one of claims 1-4 or containing a fusion protein as described in any one of claims 5-10; or, said virus-like particle is composed of a truncated Spike protein as described in any one of claims 1-4 or a fusion protein as described in any one of claims 5-10.

23. The virus-like particle of claim 22, wherein, The virus-like particles also contain nucleic acid components, which are selected from any one or more of the following: (a) Exogenous DNA, or exogenous DNA encoding exogenous proteins or polypeptides, or exogenous DNA encoding RNA; (b) A vector that optionally comprises the exogenous DNA described in (a); (c) Exogenous RNA, or exogenous RNA encoding exogenous proteins or polypeptides; (d) A vector that optionally contains the exogenous RNA described in (c).

24. The virus-like particle of claim 23, wherein, The nucleic acid component is a packaging plasmid.

25. A method for preparing a SARS-CoV-2 pseudovirus, the method comprising transferring the vector and packaging plasmid as described in any one of claims 12-14 into a host cell.

26. The method of claim 25, wherein, Transfect the vector and packaging plasmid according to any one of claims 12-14 into the host cell.

27. The method of claim 26, wherein, The mass ratio of the vector according to any one of claims 12-14 to the packaging plasmid used in the transfection process is 1-3:1-4.

28. The method of claim 26, wherein, The mass ratio of the vector according to any one of claims 12-14 to the packaging plasmid used in the transfection process is 1:

3.

29. The method of claim 26, wherein, The transfection reagents used in the transfection process are selected from: lipofectamine2000, lipofectamine3000, PEI, or any combination thereof.

30. The method of claim 25, wherein, The host cell is a human cell.

31. The method of claim 30, wherein, The cells in the human body are epithelial cells, liver cells, or tumor cells.

32. The method of claim 25, wherein, The host cell is a 293T cell that expresses or overexpresses human ACE2 protein.

33. A kit comprising one or more components selected from the following: a truncated Spike protein according to any one of claims 1-4, a fusion protein according to any one of claims 5-10, a nucleic acid molecule according to claim 11, a vector according to any one of claims 12-14, a host cell according to any one of claims 15-21, and a virus-like particle according to any one of claims 22-24.

34. The kit according to claim 33, wherein, The kit also includes packaging plasmids.

35. The kit according to claim 33, wherein, The kit also includes transfection reagents.

36. The kit of claim 35, wherein, The transfection reagent is selected from: lipofectamine2000, lipofectamine3000, PEI, or any combination thereof.

37. Use of the virus-like particles of any one of claims 22-24 in the preparation of a kit for detecting the neutralizing activity of SARS-CoV-2 antibodies, said use comprising contacting the virus-like particles with the antibody or the host cells before, simultaneously with, or after contacting the virus-like particles of any one of claims 22-24 with the host cells.

38. A method for screening candidate drugs capable of inhibiting SARS-CoV-2 infection of cells, the method comprising contacting the virus-like particles or host cells with the candidate drug before, simultaneously with, or after contacting the virus-like particles of any one of claims 22-24 with host cells.

39. Use of the truncated Spike protein of any one of claims 1-4, the fusion protein of any one of claims 5-10, the nucleic acid molecule of claim 11, the vector of any one of claims 12-14, the host cell of any one of claims 15-21, or the virus-like particle of any one of claims 22-24 in the preparation of a kit for detecting or screening antibodies or drugs against SARS-CoV-2.

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