CN116710127A

CN116710127A - Severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) polypeptides and their use for vaccine purposes

Info

Publication number: CN116710127A
Application number: CN202180059200.2A
Authority: CN
Inventors: Y·利维; G·祖拉夫斯基; S·祖拉夫斯基; C·拉卡巴拉茨; S·卡迪诺; M·苏诺
Original assignee: University Paris 12 Val De Marne; Assistance Publique Hopitaux de Paris APHP; Institut National de la Sante et de la Recherche Medicale INSERM; Baylor Research Institute
Current assignee: University Paris 12 Val De Marne; Assistance Publique Hopitaux de Paris APHP; Institut National de la Sante et de la Recherche Medicale INSERM; Baylor Research Institute
Priority date: 2020-05-26
Filing date: 2021-05-26
Publication date: 2023-09-05

Abstract

Undeniably, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has become the greatest global health threat faced by humans in this century. If the virus stabilizes the heel in the population, the SARS-CoV-2 vaccine is critical to reduce morbidity and mortality. The inventors have established a candidate vaccine against SARS-CoV-2. In particular, the inventors have identified specific epitopes to be included in candidate vaccines, as the amino acid sequences of these proteins were subjected to computer-simulated analysis by online software (NetMHC-4.0 and NetMHC II-2.3) and peptide binding prediction software to locate (map) predicted MHC-I and MHC-II epitopes. The B cell epitope was also located using on-line software (Bepippred-2.0 and discover), as well as a region enriched for epitopes of homologous sequences between SARS-CoV-2 and SARS-CoV-1. Finally, the present inventors have raised some specific CD40 antibodies that comprise one or more SARS-CoV-2 polypeptides of the invention and are suitable for vaccine purposes. Thus, the present invention relates to SARS-CoV-2 polypeptides and their use for vaccine purposes.

Description

Severe acute respiratory syndrome coronavirus 2 (SARS-COV-2) polypeptides and their use for vaccine purposes

Technical Field

The present invention relates to the field of medicine, in particular virology and vaccinology.

Background

The increased speed, rate and observed mortality of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) worldwide presents challenges to public health, socioeconomic and science. Since its transmission appears to be very active, it has been infected by more than 185 countries by 5 months and 10 days 2020, with more than 4,100,000 diagnosed cases and more than 280,000 deaths. SARS-CoV-2 can cause respiratory syndrome with clinical pathology similar to mild upper respiratory disease (common cold-like symptoms), occasionally with severe lower respiratory disease and extra-pulmonary manifestations, leading to multiple organ failure and death. This pandemic occurs following several highly pathogenic human coronavirus infections, including SARS-CoV with a mortality rate of 10% in 2002 and MERS-CoV with a mortality rate of 36% in 2012. There is no treatment or vaccine available. However, if the virus is stable in the heel of a human population, the SARS-CoV-2 vaccine is critical to reduce morbidity and mortality.

Disclosure of Invention

The present invention relates to SARS-CoV-2 polypeptide and its use for vaccine purposes.

Detailed Description

Undeniably, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has become the greatest global health threat faced by humans in this century. The speed, rate, and observed increase in mortality of global spread present challenges to public health, socioeconomic and scientific. SARS-CoV-2 can cause respiratory syndrome with clinical pathology similar to mild upper respiratory disease (common cold-like symptoms), and occasionally severe lower respiratory disease and extrapulmonary manifestations, leading to multiple organ failure and death. There are no therapeutic methods or vaccines available. However, if the virus is stable in the heel of a human population, the SARS-CoV-2 vaccine is critical to reduce morbidity and mortality. In order to accelerate the development of candidate vaccines, it is critical to use a custom and adapted ready-to-use vaccine platform for emerging and re-emerging pathogens. The inventors have established a candidate vaccine against SARS-CoV-2. In particular, the inventors have identified specific epitopes to be included in candidate vaccines, as the amino acid sequences of these proteins were subjected to computer-simulated analysis by online software (NetMHC-4.0 and NetMHC II-2.3) and peptide binding prediction software to locate (map) predicted MHC-I and MHC-II epitopes. The B cell epitopes and epitope-rich regions, which have homology between SARS-CoV-2 and SARS-CoV-1, were also mapped using on-line software (Bepippred-2.0 and discover).

Definition:

as used herein, the term "subject" or "subject in need thereof" refers to a human or non-human mammal. Typically, the patient has or is likely to be infected with SARS-CoV-2.

As used herein, the term "coronavirus" has its ordinary meaning in the art, referring to any member of the Coronaviridae family (Coronaviridae). Coronaviruses are positive-strand RNA viruses whose genomes are about 27kb to about 33kb long (depending on the particular virus). The virion RNA has a cap at the 5 'end and a polyA tail at the 3' end. The length of the RNA maximizes the size of the coronavirus in the RNA viral genome. In particular, coronavirus RNA encodes: (1) an RNA-dependent RNA polymerase; (2) N protein; (3) a tri-envelope glycoprotein; and (4) three non-structural proteins. These coronaviruses infect a wide variety of mammals and birds. They can lead to respiratory tract infections (common), intestinal infections (which occur mainly in infants >12 months), and possibly to neurological syndromes. Coronaviruses are transmitted by aerosols of respiratory secretions.

As used herein, the term "severe acute respiratory syndrome coronavirus 2" or "SARS-CoV-2" has its ordinary meaning in the art, meaning a strain of coronavirus that causes coronavirus disease 2019 (covd-19), coronavirus disease 2019 refers to respiratory syndrome, whose clinical pathology behaves similarly to mild upper respiratory disease (common cold-like symptoms), and occasionally severe lower respiratory disease and extra-pulmonary manifestations, leading to multiple organ failure and death. In particular, the term refers to Severe acute respiratory syndrome coronavirus 2 isolate 2019-nCoV_HKU-SZ-005b_2020, the complete genome of which is available under NCBI accession number MN 975262.

As used herein, the term "Covid-19" refers to respiratory disease caused by severe acute respiratory syndrome coronavirus 2.

As used herein, the term "asymptomatic" refers to a subject who has not experienced symptoms of a detectable coronavirus infection. As used herein, the term "symptomatic" refers to a subject experiencing detectable symptoms of a coronavirus infection. Symptoms of coronavirus infection include: fatigue, loss of sense of smell, headache, cough, fever, dyspnea.

As used herein, the terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to a polymer of amino acids of any length. These terms also include modified amino acid polymers; for example, disulfide bond formation, glycosylation, lipidation, phosphorylation or coupling to a labeling component. When discussed in the context of gene therapy, a polypeptide refers to the corresponding intact polypeptide, or any fragment or genetically engineered derivative thereof that retains the desired biochemical functions of the intact protein.

As used herein, the term "polynucleotide" refers to a polymeric form of nucleotides of any length, including deoxyribonucleotides or ribonucleotides, or analogs thereof. Polynucleotides may comprise modified nucleotides, such as methylated nucleotides and nucleotide analogs, and may be interrupted by non-nucleotide components. The nucleotide structure, if present, may be modified before or after assembly of the polymer. As used herein, the term polynucleotide refers interchangeably to double-stranded and single-stranded molecules. Unless otherwise indicated or required, any embodiment of the invention as described herein is a polynucleotide that includes each of the double stranded form, as well as two complementary single stranded forms known or predicted to constitute the double stranded form.

As used herein, the expression "derived from" refers to a process whereby a first component (e.g., a first polypeptide) or information from the first component is used to isolate, derive, or prepare a second, different component (e.g., a second polypeptide that is different from the first polypeptide).

As used herein, "percent identity" between two sequences is a function of the number of identical positions that both sequences have (i.e., identity% = number of identical positions/total number of positions x 100), taking into account the number of gaps that need to be introduced to achieve optimal alignment of the two sequences and the length of each gap. Sequence comparison and determination of percent identity between two sequences can be accomplished using mathematical algorithms as described below. The percent identity between two amino acid sequences can be determined using Needleman and Wunsch algorithms (Needleman, saulB. & Wunsch, christian d. (1970) "A general method applicable to the search for similarities in the amino acid sequence of two proteins", journal of Molecular biology.48 (3): 443-53.). The percent identity between two nucleotide or amino acid sequences may also be determined using an algorithm, such as EMBOSS Needle (alignment; available at www.ebi.ac.uk). For example, EMBOSS Needle may be used with BLOSUM62 matrix with a "gap opening penalty" of 10, a "gap extension penalty" of 0.5, an error "end gap penalty", an "end gap opening penalty" of 10, and an "end gap extension penalty" of 0.5. In general, "percent identity" is a function of the number of matching locations divided by the number of comparison locations, multiplied by 100. For example, if 6 are identical in 10 sequence positions between two compared sequences after alignment, the identity is 60%. % identity is typically determined over the entire length of the query sequence being analyzed. Two molecules having the same primary amino acid sequence or nucleic acid sequence are identical, irrespective of any chemical and/or biological modification. According to the invention, the first amino acid sequence having at least 90% identity to the second amino acid sequence means that the first sequence has 90 to the second amino acid sequence; 91;92;93;94;95;96;97;98;99 or 100% identity.

As used herein, the term "mutation" has its ordinary meaning in the art, referring to a substitution, deletion, or insertion. In particular, the term "substitution" refers to the removal of a particular amino acid residue at a particular position and the insertion of another amino acid residue at the same position. In the specification, mutation references are made according to standard mutation nomenclature. In particular, the term "mutation" includes "naturally occurring mutation" and "non-naturally occurring mutation".

As used herein, the term "naturally occurring mutation" refers to any mutation that can be found in a naturally occurring variant of a SARS-CoV-2 polypeptide, generally including the b.1.1.7 lineage (also known as 20I/501y.v1 variant of interest (VOC) 202012/01), the b.1.351 lineage (also known as 20H/501y.v2) and the p.1 lineage (also known as 20J/501y.v3). Such mutations are well known in the art, including those described in the references incorporated by reference below:

·(1)Jie Hu et al.The D614G mutation of SARS-CoV-2spike protein enhances viral infectivity and decreases neutralization sensitivity to individual Convalescent sera.bioRxviv(2020).

·(2)Korber B.et al.Spike mutation pipeline revcals the emergence of a more transmissible form of SARS-CoV-2.bioRxviv(2020).doi.org/10.1101/2020.04.29.069054.

·(3)Lizhou Zhang et al.The D614G mutation in the SARS-CoV-2 spike protein reduces S1 shedding and increases infectivity.bioRxxviv(2020).doi.org/10.1101/2020.06.12.148726.

·(4)Junxian Ou et al.Emergence of RBD mutations in circulating SARS-CoV-2 strains enhancing the structural stability and human ACE2 receptor affinity of the spike protcin.bioRxiv(2020).doi：10.1101/2020.03.15.991844v4

·(5)Saha，P.et al.Mutations in Spike Protcin of SARS-CoV-2 Modulate Receptor Binding，Membrane Fusion and Immunogenicity：An Insight into Viral Tropism and Pathogenesis of COVID-19.chemRxiv(2020).doi：10.26434/chemrxiv.12320567.v1

·(6)Jian Shang，Yushun Wan，Chuming Luo，Gang Ye，Qibin Geng，Ashley Auerbach，Fang Li.Cell entry mechanisms of SARS-CoV-2.Proceedings of the National Academy of Sciences May 2020，117(21)11727-11734；DoT：10.1073/pnas.2003138117

·(7)Allison J.Greaney，Andrea N.Loes，Katharine H.D.Crawford，Tyler N.Starr，Keara D.Malone，Helen Y.Chu，Jesse D.Bloom，bioRxiv 2020.12.31.425021；doi：https：//doi.org/10.1101/2020.12.31.425021

·(8)Nicholas G.Davies，Rosanna C.Barnard，Christopher I.Jarvis，Adam J.Kucharski，James Munday，Carl A.B.Pcarson，Timothy W.Russell，Damien C.Tully，Sam Abbott，Amy Gimma，William Waites，Kerry LM Wong，Kevin van Zandvoort，CMMID COVID-19 Working Group，Rosalind M.Eggo，Sebastian Funk，Mark Jit，Katherine E.Atkins，W.John Edmunds.Estimated transmissibility and severity of novel SARS-CoV-2 Variant of Concern 202012/01 in England.medRxiv 2020.12.24.20248822；doi：https：//doi.org/10.1101/2020.12.24.20248822

·(9)Houriiyah Tegally，Eduan Wilkinson，Marta Giovanetti，et al.Emergence and rapid spread of a new severe acute respiratory syndrome-related coronavirus 2(SARS-CoV-2)lineage with multiple spike mutations in South Africa.medRxiv 2020.12.21.20248640；doi：https：//doi.org/10.1101/2020.12.21.20248640

·(10)Kim JS，Jang JH，Kim JM，Chnng YS，Yoo CK，Han MG.Genome-Wide Identihcation and Characterization of Point Mutations in the SARS-CoV-2Genome.Osong Public Health Res Perspect.2020；11(3)：101-111.doi：10.24171/j.phrp.2020.11.3.05

·(11)Nilgiriwala K，Mandal A，Patel G，Mestry T，Vaswani S，Shaikh A，Sriraman K，Parikh S，Udupa S，Chatterjee N，Shastri J，Mistry N.Genome Sequences of Five SARS-CoV-2 Variants from Mumbai，India，Obtained by Nanopore Sequencing.Microbiol Resour Announc.2021 Apr 15；10(15)：e00231-21

·(12)Wenjuan Zhang，Brian D Davis，Stephanie S Chen，Jorge M Sincuir Martinez，Jasmine T Plummer，Eric Vail.Emergence of a Novel SARS-CoV-2 Variant in Southern Califomia.JAMA.2021 Apr 6；325(13)：1324-1326

for example, mutation N501Y is a nonsensical mutation in the Receptor Binding Domain (RBD) of the S protein, and is the first identified in southeast England, brazil/Japan, and south Africa, respectively, of all three SARS-CoV-2 lineages B.1.1.7, P.1 (also known as 20J/501Y.V3), and 501Y.V2. This mutation is one of the key contact residues in RBD and has been identified as increasing binding affinity to human and murine ACE 2. The E484K mutation within the Receptor Binding Domain (RBD) of the S protein (present in the new lineages 501y.s2 and b.1.1.28 from south africa and brazil, respectively) affects residues within RBD that have proven to be important for binding to many neutralizing antibodies. The E484Q mutation within the Receptor Binding Domain (RBD) of the S protein (present in new lineages b.1.617 and b.1.429 from india and denmark, respectively) also affects the same residues within RBD. Studies have shown that the L452R mutation can stabilize the interaction between spike protein and its human ACE2 receptor, thereby increasing viral infectivity. Thus, this mutation affects antibody recognition and enables SARS-CoV-2 to escape immunologically. Viruses carrying such mutations have been shown to evade recognition of antibodies in human convalescence serum, thus altering the effectiveness of the vaccine (see, e.g., allison j. Great, andrea n. Loes, katharine h. D. Crawford, tyler n. Starr, keara d. Malone, helen y. Chu, jesse d. Bloom, bioRxiv 2020.12.31.425021). Several other mutations were also found. The mutation K417N, K417T, V367F, N354D, W436R or V483A of the S1 protein has been demonstrated to have a higher affinity for binding to ACE 2. It has been previously reported that in MERS and SARS-CoV studies, the V483A and G476S mutations are associated with affinity for binding to human receptors. On the other hand, R408I may reduce the binding affinity of ACE 2. Thus, according to the present invention, the major naturally occurring mutation comprises the amino acid sequence set forth in SEQ ID NO:3 (wherein amino acid residue (K) at position 417 in SEQ ID NO:3 is replaced by amino acid residue (N), a K417N mutation in SEQ ID NO:3 (wherein amino acid residue (K) at position 417 in SEQ ID NO:3 is replaced by amino acid residue (T)), the amino acid sequence set forth in SEQ ID NO:3 (wherein amino acid residue (E) at position 484 in SEQ ID NO:3 is replaced with amino acid residue (K)), the sequence set forth in SEQ ID NO:3, wherein the amino acid residue (E) at position 484 is substituted with an amino acid residue (Q), the amino acid sequence set forth in SEQ ID NO:3 (wherein the amino acid residue (L) at position 452 in SEQ ID NO:3 is replaced by an amino acid residue (N)) and in SEQ ID NO:3 (wherein the amino acid residue (N) at position 501 in SEQ ID NO:3 is substituted with the amino acid residue (Y).

As used herein, the term "non-naturally occurring mutation" refers to any mutation inserted into a polypeptide of the invention by genetic engineering. In particular, the mutations are inserted to facilitate production of the polypeptide. For example, the mutation comprises a sequence set forth in SEQ ID NO:3 (wherein amino acid residue (C) at position 136 in SEQ ID NO:3 is replaced with amino acid residue (S)) and in SEQ ID NO:3 (wherein amino acid residue (C) at position 538 in SEQ ID NO:3 is replaced with amino acid residue (S). The mutations are particularly useful for avoiding disulfide bond formation within the polypeptides of the invention.

As used herein, the term "membrane protein" or "protein M" refers to the SARS-CoV-2 protein as a component of the viral envelope, which plays a central role in viral morphogenesis and assembly through interactions with other viral proteins. Typically, the membrane protein has the amino acid sequence set forth in SEQ ID NO:1, and a polypeptide having the amino acid sequence shown in 1.

SEQ ID NO:1 > sp|p0 dtc5|vme1_sar2 membrane protein os=severe acute respiratory syndrome coronavirus 2 OX = 2697049 PE =3 SV =1. The polypeptides Mpep1 and Mpep2 are underlined.

MADSNGTITVEELKKLLEQWNLVIGFLFLTWICLLQFAYANRNRFLYIIKLIFLWLLWPVTLACFVLAA VYRINWITGGIAIAMACLVGLMWLSYFIASFRLFARTRSMWSFNPETNILLNVPLHGTILTRPLLESELVIGAVILR GHLRIAGHHLGRCDIKDLPKEITVATSRTLSYYKLGASQRVAGDSGFAAYSRYRIGNYKLNTDHSSSSDNIALLVQ

As used herein, the term "nucleoprotein" or "protein N" refers to the SARS-CoV-2 protein that packages the orthochain viral genomic RNA into a helical Ribonucleocapsid (RNP) and plays a fundamental role in viral particle assembly through the interaction of this protein with the viral genome and membrane protein M. Typically, the nucleoprotein has the sequence set forth in SEQ ID NO:2, and a polypeptide having the amino acid sequence shown in 2.

SEQ ID NO:2 > sp|p0 dtc9|ncap_sars2nucleoprotein os=severe acute respiratory syndrome

Coronavirus 2 OX = 2697049 GN =npe=1 SV =1. The polypeptides Npep1 and Npep2 are underlined.

MSDNGPQNQRNAPRITFGGPSDSTGSNQNGERSGARSKQRRPQGLPNNTASWFTALTQHGKEDLKFPRGQGVPINTNSSPDDQIGYYRRATRRIRGGDGKMKDLSPRWYFYYLGTGPEAGLPYGANKDGIIWVATEGALNTPKDHI GTRNPANNAAIVLQLPQGTTLPKGFYAEGSRGGSQASSRSSSRSRNSSRNSTPGSSRGTSPARMAGNGGDAALALLLLDRLNQLESKMSGKGQQQQGQTVTKKSAAEASKKPRQKRTATKAYNVTQAFGRRGPEQTQGNFGDQELIRQGTDYKH WPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKAD ETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMSSADSTQA

As used herein, the term "spike protein" or "protein S" refers to the SARS-CoV-2 spike glycoprotein that binds to its cellular receptor (i.e., ACE 2) and mediates membrane fusion and viral entry. Each monomer of the trimeric S protein is approximately 180 kDa and comprises two subunits S1 and S2, which mediate attachment and membrane fusion, respectively. In particular, spike protein S1 attaches the viral particle to the cell membrane by interaction with a host receptor (i.e., the human ACE2 receptor). Spike protein S2 mediates fusion of virions and cell membranes by acting as a class I viral fusion protein. In the current model, the protein has at least three conformational states: a natural state before fusion, an intermediate state before hairpin and a hairpin state after fusion. During fusion of the viral and target cell membranes, the coiled-coil region (heptad repeat) assumes a hairpin trimer structure, localizing the fusion peptide to the C-terminal region near the extracellular domain. The formation of this structure appears to drive the juxtaposition (appposition) and subsequent fusion of the viral and target cell membranes. Spike protein S2' functions as a viral fusion peptide that is exposed after S2 cleavage upon endocytosis of the virus (unmask). Typically, the spike protein has the amino acid sequence as set forth in SEQ ID NO:3, and a polypeptide having the amino acid sequence shown in 3.

SEQ ID NO:3 > sp|p0dtc2|spike_sars2 SPIKE glycoprotein os=severe acute respiratory syndrome coronavirus 2 OX = 2697049 GN =sspe=1 SV =1. The polypeptides Spep1, spep2, spep3 and Spep4 are underlined. RBD polypeptides are shown in italics and bold.

VAIHADQLTPTWRVYSTGSNVFQTRAGCLIGAEHVNNSYECDIPIGAGICASYQTQTNSPRRARSVASQ SIIAYTMSLGAENSVAYSNNSIATPTNFTISVTTETLPVSMTKTSVDCTMYTCGDSTECSNLLLQYGSFCTQLNRAL TGIAVEQDKNTQEVFAQVKQIYKTPPIKDFGGFNFSQILPDPSKPSKRSFIEDLLFNKVTLADAGFIKQYGDCLGDI AARDLICAQKFNGLTVLPPLLTDEMIAQYTSALLAGTITSGWTFGAGAALQIPFAMQMAYRFNGIGVTQNVLYENQK LIANQFNSAIGKIQDSLSSTASALGKLQDVVNQNAQALNTLVKQLSSNFGAISSVLNDILSRLDKVEAEVQIDRLIT GRLQSLQTYVTQQLIRAAEIRASANLAATKMSECVLGQSKRVDFCGKGYHLMSFPQSAPHGVVFLHVTYVPAQEKNF TTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEEL DKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQYIKWPWYIWLGFIAGLIAIVMVTIMLCCMTSCCSCLKGCCSCGSCCKFDEDDSEPVLKGVKLHYT

As used herein, the term "RBD polypeptide" refers to a polypeptide consisting of the amino acid sequence: and the sequence of SEQ ID NO:3 (i.e., in a spike protein), the amino acid sequence ranging from amino acid residue 319 to amino acid residue 541 has at least 90% identity.

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541.

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises one or more non-naturally occurring mutations. In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a non-naturally mutated amino acid at position 538. In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a C538S mutation.

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises one or more naturally occurring mutations. In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises one or more naturally occurring mutations at positions 417, 452, 484 or 501. In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprising one or more naturally occurring mutations in a position selected from the group consisting of the K417N, K417T, L452N, E84Q, E K and N501Y mutations.

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises one or more non-naturally occurring mutations, in particular a C538S mutation, and one or more naturally occurring mutations, preferably one or more of the K417N, K417T, L452N, E84Q, E484K and N501Y mutations.

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 amino acid residue at position 319

Amino acid composition ranging from amino acid residues at positions 541 and comprising naturally occurring mutations of N501Y and non-naturally occurring mutations of C538S.

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a naturally occurring mutation of K417T, E484K, N Y and a non-naturally occurring C538S mutation.

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a naturally occurring mutation of K417T, E484K, N501Y and a non-naturally occurring C538S mutation(“RBD _{South Africa variants} "OR" RBD _SA _Var ”)。

In some embodiments, the RBD polypeptide consists of the amino acid sequence set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises the naturally occurring mutation of E484Q, L452N and the non-naturally occurring C538S mutation ("RBD) _{Indian variants} ”)。

As used herein, the term "conjugate" or interchangeably "conjugated polypeptide" is intended to mean a combination or chimeric molecule formed by covalent attachment of one or more polypeptides. The term "covalently linked" or "coupled" refers to the covalent attachment of polypeptide and non-peptide moieties to each other either directly or indirectly through one or more spacer moieties (e.g., a bridge, spacer, or one or more bond moieties). A particular conjugate is a fusion protein.

As used herein, the term "fusion protein" comprises at least one polypeptide of the invention operably linked to a heterologous polypeptide. In fusion proteins, the term "operably linked" means that the peptide of the invention and the heterologous polypeptide are fused in-frame to each other.

As used herein, the term "heterologous polypeptide" refers to a polypeptide that is not derived from the same protein as the heterologous polypeptide fused to.

As used herein, the term "linker" refers to at least one sequence of amino acids that links a peptide of the invention to a heterologous polypeptide. Linkers are well known to those of ordinary skill in the art and typically comprise 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 or more amino acids.

As used herein, the term "antibody" refers to immunoglobulin molecules and immunologically active portions of immunoglobulin molecules, i.e., molecules that contain an antigen binding site that immunospecifically binds to an antigen. In natural antibodies in rodents and primates, two heavy chains are linked to each other by disulfide bonds, each heavy chain being linked to a light chain by disulfide bonds. There are two types of light chains, λ (1) and κ (k). There are five main heavy chain classes (or isotypes) that determine the functional activity of an antibody molecule: igM, igD, igG, igA and IgE. Each chain comprises a different sequence domain. In a typical IgG antibody, the light chain comprises two domains, a variable domain (VL) and a constant domain (CL). The heavy chain comprises four domains, one variable domain (VH) and three constant domains (CH 1, CH2 and CH3, collectively referred to as CH). The variable regions of the light chain (VL) and heavy chain (VH) determine the binding recognition and specificity for an antigen. The constant region domains of the light Chain (CL) and heavy Chain (CH) confer important biological properties such as antibody chain attachment, secretion, trans-placental flow (trans-placental mobility), complement binding, and binding to Fc receptors (FcR). Fv fragments are the N-terminal part of the Fab fragment of an immunoglobulin, consisting of the variable parts of one light chain and one heavy chain. The specificity of an antibody is due to the structural complementarity between the binding site of the antibody and the epitope. The antibody binding site consists of residues primarily from the hypervariable region or Complementarity Determining Regions (CDRs). Sometimes, residues from non-hypervariable regions or Framework Regions (FR) may participate in the antibody binding site, or affect the overall domain structure, thereby affecting the binding site. Complementarity determining regions or CDRs refer to amino acid sequences that collectively define the binding affinity and specificity of the native Fv region of the native immunoglobulin binding site. The light and heavy chains of an immunoglobulin each have three CDRs, which are referred to as L-CDR1, L-CDR2, L-CDR3 and H-CDR1, H-CDR2, H-CDR3, respectively. Thus, an antigen binding site typically comprises six CDRs, including sets of CDRs from each of the heavy and light chain V regions. The Framework Region (FR) refers to the amino acid sequence between the CDRs. Thus, the variable regions of the light and heavy chains typically comprise 4 framework regions and 3 CDRs in the following order: FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4. Residues in the antibody variable domains are typically numbered according to the system designed by Kabat et al. See Kabat et al 1987,in Sequences of Proteins ofImmunological Interest,US Department of Health and Human Services,NIH,USA (Kabat et al 1992, hereinafter "Kabat et al"). The Kabat residue name does not always correspond directly to the linear numbering of amino acid residues in the SEQ ID sequence. The actual linear amino acid sequence may contain fewer or additional amino acids than in the strict Kabat numbering, corresponding to a shortened structural component or an inserted structural component in the basic variable domain structure, whether framework or Complementarity Determining Regions (CDRs). For a given antibody, the correct Kabat residue number can be determined by aligning homologous residues in the antibody sequence to a "standard" Kabat numbering sequence. According to the Kabat numbering system, the CDRs of the heavy chain variable domain are located at residues 31-35 (H-CDR 1), residues 50-65 (H-CDR 2) and residues 95-102 (H-CDR 3). According to the Kabat numbering system, the CDRs of the light chain variable domain are located at residues 24-34 (L-CDR 1), residues 50-56 (L-CDR 2) and residues 89-97 (L-CDR 3). For the antibodies described below, CDR-see section of the antibody page titled How to identify the CDRs by looking at a sequence has been determined using the CDR-finding algorithm from www.bioinf.org.uk.

As used herein, the term "immunoglobulin domain" refers to a globular region of an antibody chain (e.g., a chain of a conventional 4-chain antibody, or a chain of a heavy or light chain antibody), or to a polypeptide consisting essentially of such a globular region.

As used herein, the term "Fc region" is used to define the C-terminal region of an immunoglobulin heavy chain, including native sequence Fc regions and variant Fc regions. The human IgG heavy chain Fc region is generally defined as comprising amino acid residues from position C226 or from position P230 to the carboxy terminus of an IgG antibody. The residue number of the Fc region is the EU index of Kabat. The C-terminal lysine (residue K447) of the Fc region may be removed, for example, during production or purification of the antibody. Thus, an antibody composition of the invention may comprise a population of antibodies that have all K447 residues removed, a population of antibodies that have no K447 residues removed, and a population of antibodies that have a mixture of antibodies with or without K447 residues.

As used herein, the term "epitope" has its ordinary meaning in the art, being a fragment of at least 8 amino acids that is recognized by an immunoreactive component. As used herein, the term "immunoreactive component" includes, but is not limited to, macrophages, lymphocytes, T lymphocytes, killer T lymphocytes, immune response modifiers, helper T lymphocytes, antigen receptors, antigen presenting cells, cytotoxic T lymphocytes, T-8 lymphocytes, CD1 molecules, B lymphocytes, antibodies, recombinant antibodies, genetically engineered antibodies, chimeric antibodies, monospecific antibodies, bispecific antibodies, multispecific antibodies, diabodies, chimeric antibodies, humanized antibodies, human antibodies, heterologous antibodies, monoclonal antibodies, polyclonal antibodies, antibody fragments, and/or at least a portion of a synthetic antibody. The term "epitope" may be used interchangeably with antigen, paratope binding site, antigenic determinant and/or determinant.

As used herein, the term "multi-epitope polypeptide" refers to a polypeptide comprising at least 2 epitopes. In particular, the polyepitope polypeptides of the invention comprise one or more SARS-CoV-2 polypeptides of the invention.

As used herein, the term "antibody epitope" refers to a peptide that can be recognized by, or induce the formation of, a specific antibody.

As used herein, the term "chimeric antibody" refers to an antibody comprising VH and VL domains of a non-human antibody, and CH and CL domains of a human antibody. In one embodiment, a "chimeric antibody" is an antibody molecule in which (a) the constant region (i.e., heavy and/or light chain) or portions thereof are altered, substituted, or exchanged such that the antigen binding site (variable region) is linked to a different or altered class, effector function, and/or class of constant region, or an entirely different molecule (e.g., enzyme, toxin) that confers novel properties to the chimeric antibody of an agonist molecule (e.g., CD40 ligand, hormone, growth factor, drug, etc.); or (b) the variable region or portion thereof is altered, replaced or exchanged with a variable region having a different or altered antigen specificity. Chimeric antibodies also include primatized antibodies, particularly humanized antibodies. Furthermore, the chimeric antibody may comprise residues not found in the recipient antibody or the donor antibody. These modifications were made to further improve antibody performance. See Jones et al, nature 321 for details: 522-525 (1986); riechmann et al, nature 332:323-329 (1988); and Presta, curr.Op.struct.biol.2:593-596 (1992). (see U.S. Pat. No. 4,816,567; and Morrison et al, proc. Natl. Acad. Sci. USA,81:6851-6855 (1984)).

As used herein, the term "humanized antibody" includes antibodies having 6 CDRs of a murine antibody but humanized framework and constant regions. More specifically, the term "humanized antibody" as used herein may include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) have been grafted to human framework sequences.

As used herein, the term "human monoclonal antibody" is intended to include antibodies having variable and constant regions derived from human immunoglobulin sequences. The human antibodies of the invention may include amino acid residues not encoded by human immunoglobulin sequences (e.g., mutations introduced by random or site-directed mutagenesis in vitro, or by somatic mutation in vivo). However, in one embodiment, the term "human monoclonal antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) have been grafted to human framework sequences.

As used herein, the term "human leukocyte antigen system" or "HLA" has its ordinary meaning in the art, referring to the Major Histocompatibility Complex (MHC) in humans. The locus comprises a plurality of genes encoding cell surface antigen presenting proteins. Proteins encoded by certain genes are also referred to as antigens. The major HLA antigens are HLA class I antigens (A, B and C) and HLA class II antigens (DR, DP and DQ). HLA class I antigens present peptides (8-12 amino acids) that are usually derived from the interior of the cell, attracting CD8 cytotoxic T cells that destroy the cell. HLA class II antigens present peptides, which are usually derived from external cells, to CD 4T helper lymphocytes, thereby stimulating B cells and other immune cells.

As used herein, the term "immune response" refers to a response of the immune system to an antigen in a host, including the production of antigen-specific antibodies and/or cytotoxic responses. For primary antigen exposure an immune response (primary immune response) is usually detectable after a lag period of several days to two weeks; the immune response to subsequent stimulation of the same antigen (secondary immune response) is more rapid than the primary immune response. Immune responses to the transgene product may include humoral (e.g., antibody responses) and cellular (e.g., cytolytic T cell responses) immune responses, which may be caused by the immunogenic product encoded by the transgene. The level of immune response may be measured by methods known in the art (e.g., by measuring antibody titers).

As used herein, the term "APC" or "antigen presenting cell" refers to a cell capable of activating T cells, including but not limited to certain macrophages, B cells, and dendritic cells.

As used herein, the term "dendritic cell" or "DC" refers to any member of a different population of morphologically similar cell types found in lymphoid or non-lymphoid tissues. These cells are characterized by their unique morphology, high surface level expression of MHC class II (Steinman et al, ann. Rev. Immunol.9:271 (1991); the description of which is incorporated herein by reference).

As used herein, the term "CD40" has its ordinary meaning in the art, referring to the human CD40 polypeptide receptor. In some embodiments, CD40 is an isoform of the human canonical sequence reported by UniProtKB-P25942 (also known as human TNR 5).

As used herein, the term "CD40L" has its ordinary meaning in the art, referring to a human CD40L polypeptide, e.g., as reported by UniProtKB-P25942, including its SEQ ID NO:4, a CD40 binding domain. CD40L may be expressed as a soluble polypeptide, a natural ligand for the CD40 receptor.

SEQ ID NO:4 > CD40L binding domain

MQKGDQNPQIAAHVISEASSKTTSVLQWAEKGYYTMSNNLVTLENGKQLTVKRQGLYYIYAQVTFCSNREASSQAPEIASLCLKSPGREERILLRAAMTHSSAKPCGQQSIHLGGVFELQPGASVFVNVTDPSQVSHGTGFTSFGLLKL

As used herein, the term "CD40 agonist antibody" refers to an antibody that increases CD 40-mediated signaling activity in the absence of CD40L in a cell-based assay (e.g., a B cell proliferation assay). In particular, CD40 agonist antibodies (i) induce B cell proliferation, as measured in vitro by flow cytometry analysis, or by repeated dilution analysis of CFSE-labeled cells; and/or (ii) induce secretion of cytokines (e.g., IL-6, IL-12, or IL-15), as measured in vitro using a dendritic cell activation assay.

As used herein, the term "langerhans element (Langerin)" has its ordinary meaning in the art, referring to a human C-type lectin domain family 4 member K polypeptide. In some embodiments, langerhans is an isoform of the human canonical sequence reported by UniProtKB-Q9UJ71 (also known as human CD 207).

As used herein, the term "treatment" or "treatment" refers to prophylactic or preventative (prophoric) treatment, as well as curative or disease modifying treatment, including treatment of patients at risk of or suspected of having an infectious disease, as well as patients ill or diagnosed as having a disease or medical condition, including inhibition of clinical recurrence. Treatment may be administered to a patient suffering from a medical condition or ultimately available condition to prevent, cure, delay the onset of, reduce the severity of, or ameliorate one or more symptoms of the condition or recurrence of the condition, or to extend the patient's survival beyond that expected without such treatment. "treatment regimen" refers to a mode of treatment for a disease, such as a mode of administration used during treatment. Treatment protocols may include induction protocols and maintenance protocols. The phrase "induction regimen" or "induction phase" refers to a treatment regimen (or portion of a treatment regimen) for the initial treatment of a disease. The overall goal of an induction regimen is to provide high levels of drug to the patient during the initial stages of the treatment regimen. The induction regimen may employ (partially or fully) a "loading regimen" which may include administration of a greater dose of the drug than the physician employs during the maintenance regimen, administration of the drug more frequently than the physician does during the maintenance regimen, or both. The phrase "maintenance regimen" or "maintenance phase" refers to a treatment regimen (or portion of a treatment regimen) for maintaining a patient during treatment of a disease, for example, to have the patient in remission for a prolonged period (month or year). The maintenance regimen may employ continuous therapy (e.g., administration of drugs at regular intervals, e.g., weekly, monthly, yearly, etc.), or intermittent therapy (e.g., discontinuing therapy, intermittent therapy, therapy upon recurrence, or therapy when certain predetermined criteria are met [ e.g., pain, disease manifestation, etc.).

As used herein, the term "pharmaceutical composition" refers to a composition as described herein or a pharmaceutically acceptable salt thereof, as well as other agents, such as carriers and/or excipients. The pharmaceutical compositions provided herein generally include a pharmaceutically acceptable carrier.

As used herein, the term "pharmaceutically acceptable carrier" includes any and all solvents, diluents or other liquid vehicles, dispersing or suspending aids, surfactants, isotonic agents, thickening or emulsifying agents, preservatives, solid binders, lubricants and the like which are suitable for the particular dosage form desired. Remington's pharmaceutical-Sciences, sixteenth Edition, e.w. martin (Mack Publishing co., easton, pa., 1980) discloses various carriers for formulating pharmaceutical compositions and known techniques for preparing the same.

As used herein, the term "vaccination" or "vaccination" refers to, but is not limited to, the process of eliciting an immune response against a particular antigen in a subject.

As used herein, the term "vaccine composition" refers to a composition that can be administered to a human or animal to induce an immune system response; this immune system response can lead to the activation of certain cells, particularly APC, T lymphocytes and B lymphocytes.

As used herein, the term "antigen" refers to a molecule that, if processed and presented by an MHC molecule, is capable of being specifically bound by an antibody or T Cell Receptor (TCR). Antigens can also be recognized by the immune system and/or can induce humoral and/or cellular immune responses, resulting in B and/or T lymphocyte activation. An antigen may have one or more epitopes or antigenic sites (B and T epitopes).

As used herein, the term "adjuvant" refers to a compound that can induce and/or enhance an immune response against an antigen when administered to a subject or animal. Adjuvants are also intended to mean substances whose action is generally that of accelerating, prolonging or enhancing the specific immune response to a specific antigen. In the context of the present invention, the term "adjuvant" refers to a compound that enhances the innate immune response by affecting the transient response of the innate immune response and that enhances the longer term effects of the adaptive immune response by activation and maturation of Antigen Presenting Cells (APCs), particularly Dendritic Cells (DCs).

As used herein, the expression "therapeutically effective amount" refers to a sufficient amount of the active ingredient of the present invention to induce an immune response at a reasonable benefit/risk ratio suitable for medical treatment.

As used herein, the term "immune checkpoint inhibitor" has its ordinary meaning in the art, referring to any compound that inhibits the function of an immune checkpoint protein. As used herein, the term "immune checkpoint protein" has its ordinary meaning in the art, referring to a molecule expressed by a T cell that either turns on a signal (stimulatory checkpoint molecule) or turns off a signal (inhibitory checkpoint molecule). Immune checkpoint molecules are believed in the art to constitute immune checkpoint pathways similar to CTLA-4 and PD-1 dependent pathways (see, e.g., pardoll,2012.Nature Rev Cancer 12:252-264; mellman et al 2011.Nature 480:480-489). Examples of inhibitory checkpoint molecules include A2AR, B7-H3, B7-H4, BTLA, CTLA-4, CD277, IDO, KIR, PD-1, LAG-3, TIM-3 and VISTA.

The polypeptide of the invention:

one object of the present invention relates to SARS-CoV-2 polypeptide (M1-110, also referred to as "Mpep 1") derived from the M protein, which consists of a polypeptide sequence identical to SEQ ID NO:1 from residue 1 to residue 110, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

In some embodiments, the polypeptide (M1-110) comprises a sequence that hybridizes to SEQ ID NO:1 from residue 1 to residue 110, 50 of the amino acid sequences having at least 90% identity to the amino acid sequence of seq id no; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109; or 110 consecutive amino acids.

In some embodiments, the polypeptide (M1-110) consists of a polypeptide that hybridizes to SEQ ID NO:1 from residue 1 to residue 110. In some embodiments, the polypeptide (M1-110) comprises one or more naturally occurring mutations.

Another object of the invention relates to the SARS-CoV-2 polypeptide (M132-222 also referred to as "Mpep 2") derived from protein M, which consists of a polypeptide sequence identical to SEQ ID NO:1 from residue 132 to residue 222, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

In some embodiments, the polypeptide (M132-222) comprises a sequence that hybridizes to SEQ ID NO:1 from residue 132 to residue 222, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90; or 91 consecutive amino acids.

In some embodiments, the polypeptide (M132-222) consists of a polypeptide that hybridizes to SEQ ID NO:1 from residue 132 to residue 222, and an amino acid sequence having at least 90% identity. In some embodiments, the polypeptide (M132-222) comprises one or more naturally occurring mutations.

Another object of the invention relates to a SARS-CoV-2 polypeptide derived from protein N (N78-206 is also referred to as "Npep 1") consisting of a polypeptide sequence that hybridizes to SEQ ID NO:2 from residue 78 to residue 206, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

In some embodiments, the polypeptide (N78-206) comprises a sequence that hybridizes to SEQ ID NO:2 from residue 78 to residue 206, 50 of the amino acid sequences having at least 90% identity to the amino acid sequence of seq id no; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. Or 129 consecutive amino acids.

In some embodiments, the polypeptide (N78-206) consists of a polypeptide sequence that hybridizes to SEQ ID NO:2 from residue 78 to residue 206. In some embodiments, the polypeptide (N78-206) comprises one or more naturally occurring mutations.

Another object of the invention relates to a SARS-CoV-2 polypeptide derived from protein N (N276-411 also known as "Npep 2") consisting of a polypeptide having a sequence identical to SEQ ID NO:2 from residue 276 to residue 411, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

In some embodiments, the polypeptide (N276-411) comprises a sequence that hybridizes to SEQ ID NO:2 from residue 276 to residue 411; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135; or 136 consecutive amino acids.

In some embodiments, the polypeptide (N276-411) consists of a polypeptide that hybridizes to SEQ ID NO:2 from residue 276 to residue 411. In some embodiments, the polypeptide (N276-411) comprises one or more naturally occurring mutations.

Another object of the invention relates to a SARS-CoV-2 polypeptide (S125-250, also known as "Spep 1") derived from protein S, which consists of a polypeptide sequence identical to SEQ ID NO:3 from residue 125 to residue 250, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

In some embodiments, the polypeptide (S125-250) comprises a sequence that hybridizes to SEQ ID NO:3 from residue 125 to residue 250, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125; or 126 consecutive amino acids.

In some embodiments, the polypeptide (S125-250) consists of a polypeptide that hybridizes to SEQ ID NO:3 from residue 125 to residue 250.

In some embodiments, the polypeptide (S125-250) comprises one or more naturally occurring mutations.

In some embodiments, the polypeptide (S125-250) consists of the sequence set forth in SEQ ID NO:3 to residue 250, and comprising a non-naturally occurring mutation at position 136. In some, the polypeptide (S125-250) consists of the amino acid sequence set forth in SEQ ID NO:3 to residue at position 250, and comprises a C136S non-naturally occurring mutation ("Spep 1) _C136S ") amino acid sequence.

Another object of the invention relates to a SARS-CoV-2 polypeptide (S280-598, also known as "Spep 2") derived from protein S, which is identical to a polypeptide consisting of a polypeptide having a sequence as set forth in SEQ ID NO:3 from residue 280 to residue 598.

In some embodiments, the polypeptide (S280-598) comprises a sequence that hybridizes to SEQ ID NO: 50 of the amino acid sequence having at least 90% identity to the amino acid sequence ranging from residue 280 to residue 598 in 3; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135;136;137, respectively; 138;139, respectively; 140;141;142;143, a base; 144 (144); 145, respectively; 146;147;148, a step of selecting a key; 149;150;151;152;153;154;155;156, respectively; 157 (157); 158;159;160, a step of detecting a position of the base; 161;162;163;164, a step of detecting the position of the base; 165;166;167;168;169 (169); 170, a step of; 171;172;173, a part of the base; 174;175;176;177 (step 1); 178;179;180;181;182;183 (V); 184;185;186, a step of detecting the position of the base; 187;188;189;190;191; 192. 193;194, a step of receiving a signal; 195;196;197;198, a step of; 199;200;201;202;203, a base station; 204; 205. 206;207 (x, y); 208;209;210;211;212;213;214;215, respectively; 216;217;218;219, a step of; 220;221;222, a step of; 223, a third step; 224;225;226;227;228;229;230, a step of; 231;232;233;234;235;236;237; 238. 239;240, a step of; 241, a base; 242;243;244;245; 246. 247, respectively; 248;249;250;251; 252. 253;254, a base plate; 255, respectively; 256;257;258;259;260;261;262;263;264;265;266;267;268, a step of; 269;270; 271(s); 272;273;274;275;276, respectively; 277;278;279;280; 281. 282;283;284;285;286;287;288;289;290;291 of a metal strip; 292;293;294;295;296;297;298;299;300;301;302;303;304;305; 306. 307. 308. 309, and (c) a third party; 310;311;312;313;314, a step of; 315;316, a step of; 317. 318; or 319 consecutive amino acids.

In some embodiments, the polypeptide (S280-598) consists of a polypeptide sequence that hybridizes to SEQ ID NO:3 from residue 280 to residue 598.

In some embodiments, the polypeptide (S280-598) consists of a polypeptide sequence that hybridizes to SEQ ID NO:3 from residue 280 to residue 598, and comprising one or more non-naturally occurring mutations. In some embodiments, the polypeptide (S280-598) consists of a polypeptide sequence that hybridizes to SEQ ID NO:3 from residue 280 to residue 598, and comprising a non-natural mutation at position 538. In some embodiments, the polypeptide (S280-598) consists of a polypeptide sequence that hybridizes to SEQ ID NO:3 from residue 280 to residue 598, and comprising a non-naturally occurring mutation of C538S.

In some embodiments, the polypeptide (S280-598) consists of the amino acid sequence set forth in SEQ ID NO:3 from residue 280 to residue 598, and comprising one or more naturally occurring mutations. In some embodiments, the polypeptide (S280-598) consists of the amino acid sequence set forth in SEQ ID NO:3 from residue 280 to residue 598, and comprising one or more naturally occurring mutations at positions 417, 438 or 501. In some embodiments, the polypeptide (S280-598) consists of the amino acid sequence set forth in SEQ ID NO:3 from residue 280 to residue 598, and comprising one or more naturally occurring mutations selected from the group consisting of the K417N, K417T, E K and N501Y mutations.

In some embodiments, the polypeptide (S280-598) consists of the amino acid sequence set forth in SEQ ID NO:3 from residue 280 to residue 598, and comprises an N501Y naturally occurring mutation and a C538S non-naturally occurring mutation.

In some embodiments, the polypeptide (S280-598) consists of the amino acid sequence set forth in SEQ ID NO:3 from residue 280 to residue 598, and comprises a naturally occurring mutation of K417T, E484K, N501Y and a non-naturally occurring mutation of C538S.

In some embodiments, the polypeptide (S280-598) consists of the amino acid sequence set forth in SEQ ID NO:3 from residue 280 to residue 598, and comprises a naturally occurring mutation of K417N, E484K, N501Y and a non-naturally occurring mutation of C538S.

Another object of the invention relates to the SARS-CoV-2 polypeptide (S680-1029, also known as "Spep 3") derived from protein S, which consists of a polypeptide sequence identical to SEQ ID NO:3 from residue 680 to residue 1029, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity.

In some embodiments, the polypeptide (S680-1029) comprises a sequence identical to SEQ ID NO: 50 of the amino acid sequence having at least 90% identity to the amino acid sequence ranging from residue 680 to residue 1029 in 3; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135;136;137, respectively; 138;139, respectively; 140;141;142;143, a base; 144 (144); 145, respectively; 146;147;148, a step of selecting a key; 149;150;151;152;153;154;155;156, respectively; 157 (157); 158;159;160, a step of detecting a position of the base; 161;162;163;164, a step of detecting the position of the base; 165;166;167;168;169 (169); 170, a step of; 171;172;173, a part of the base; 174;175;176;177 (step 1); 178;179;180;181;182;183 (V); 184;185;186, a step of detecting the position of the base; 187;188;189;190;191; 192. 193;194, a step of receiving a signal; 195;196;197;198, a step of; 199;200;201;202;203, a base station; 204; 205. 206;207 (x, y); 208;209;210;211;212;213;214;215, respectively; 216;217;218;219, a step of; 220;221;222, a step of; 223, a third step; 224;225;226;227;228;229;230, a step of; 231;232;233;234;235;236;237; 238. 239;240, a step of; 241, a base; 242;243;244;245; 246. 247, respectively; 248;249;250;251; 252. 253;254, a base plate; 255, respectively; 256;257;258;259;260;261;262;263;264;265;266;267;268, a step of; 269;270; 271(s); 272;273;274;275;276, respectively; 277;278;279;280; 281. 282;283;284;285;286;287;288;289;290;291 of a metal strip; 292;293;294;295;296;297;298;299;300;301;302;303;304;305; 306. 307. 308. 309, and (c) a third party; 310;311;312;313;314, a step of; 315;316, a step of; 317. 318;319; 320. 321, a base; 322;323 (323); 324, a base; 325, a step of; 326, a step of; 327, respectively; 328. 329;330;331;332;333;334;335;336, a base; 337, respectively; 338;339;340 (340); 341;342;343;344;345, a frame structure; 346;347;348;349; or 350 contiguous amino acids.

In some embodiments, the polypeptide (S680-1029) consists of a polypeptide sequence that hybridizes to SEQ ID NO:3 from residue 680 to residue 1029. In some embodiments, the polypeptide (S680-1029) comprises one or more naturally occurring mutations. Another object of the invention relates to the SARS-CoV-2 polypeptide (S1056-1209, also known as "Spep 4") derived from protein S, which consists of a polypeptide sequence identical to SEQ ID NO:3 from residue 1056 to residue 1209, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

In some embodiments, the polypeptide (S1056-1209) comprises a sequence that hybridizes to SEQ ID NO:3 from residue 1056 to residue 1209, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135;136;137, respectively; 138;139, respectively; 140;141;142;143, a base; 144 (144); 145, respectively; 146;147;148, a step of selecting a key; 149;150;151;152;153; or 154 consecutive amino acids.

In some embodiments, the polypeptide (S1056-1209) consists of a polypeptide sequence that hybridizes to SEQ ID NO:3 from residue 1056 to residue 1209. In some embodiments, the polypeptide (S10565-1209) comprises one or more naturally occurring mutations.

Conjugates and fusion proteins

In some embodiments, the invention relates to conjugates, wherein a heterologous polypeptide is conjugated or fused to one or more SARS-CoV-2 polypeptides of the invention.

In some embodiments, a conjugate of the invention comprises 2, 3, 4, 5, 6, 7, or 8 SARS-CoV-2 polypeptides of the invention. In some embodiments, conjugates of the invention comprise a polypeptide (M1-110, "Mpep 1"), a polypeptide (M132-222, "Mpep 2"), a polypeptide (N78-206, "Npep 1"), a polypeptide (N276-411, "Npep 2"), a polypeptide (S125-250, "Spep 1"), a polypeptide (S280-598, "Spep 2"), a polypeptide (S680-1029, "Spep 3"), and a polypeptide (S1056-1209, "Spep 4").

In some embodiments, the conjugates of the invention comprise one or more sequences derived from a restriction cloning site present in a polynucleotide encoding the conjugate. Typically, the sequence may consist of 2 amino acid residues, typically including AP, AS, AR, PR, SA, TR and TS sequences.

In some embodiments, the conjugates of the invention comprise a sequence of a signal peptide. As used herein, the term "signal peptide" has its ordinary meaning in the art, and refers to a propeptide that exists as an N-terminal peptide on a protein precursor form. The function of the signal peptide is to facilitate translocation of the expressed polypeptide to which it is linked into the endoplasmic reticulum. During this process, the signal peptide is typically cleaved. The signal peptide may be heterologous or homologous to the organism used to produce the polypeptide.

In some embodiments, the conjugates of the invention comprise a polypeptide (N276-411 "Npep 2"), a polypeptide (S125-250, "Spep 1"), and a polypeptide (S1056-1209, "Spep 4"). In some embodiments, the conjugate comprises a multi-epitope polypeptide comprising a fusion (from N-terminus to C-terminus) of a polypeptide (N276-411 "Npep 2"), a polypeptide (S125-250, "Spep 1"), and a polypeptide (S1056-1209, "Spep 4"), and optionally one or more linkers as described below (separating Npep2 from Spep1 and/or Spep1 from Spep 4). In some embodiments, the conjugate comprises a multi-epitope polypeptide having the formula Npep2-Spep1-f2-Spep4, wherein f2 represents a linker as described below. In some embodiments, the conjugate comprises a polypeptide having a sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

SEQ ID NO：5>Multi-epitope polypeptides

In some embodiments, the conjugates of the invention comprise a polypeptide (S125-250, "Spep 1") and a polypeptide (S1056-1209, "Spep 4"). In some embodiments, the conjugate comprises a multi-epitope polypeptide comprising a fusion (from N-terminus to C-terminus) of a polypeptide (N276-411 "Npep 2") and a polypeptide (S125-250, "Spep 1") and optionally a linker separating Npep2 from Spep1 as described below. In some embodiments, the conjugate comprises a multi-epitope polypeptide having the formula Spep1-f2-Spep4, wherein f2 represents a linker as described herein. In some embodiments, the conjugate comprises a polypeptide having a sequence as set forth in SEQ ID NO:6, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

SEQ ID NO：6>

In some embodiments, the conjugates of the invention comprise a polypeptide (N276-411, "Npep 2") and an RBD polypeptide. In some embodiments, the conjugate comprises a multi-epitope polypeptide comprising a fusion of a polypeptide (N276-411 "Npep 2) and an RBD polypeptide (from N-terminus to C-terminus) and optionally a linker separating Npep2 from RBD as described below. In some embodiments, the conjugate comprises a polypeptide having a sequence as set forth in SEQ ID NO: 7.

SEQ ID NO：7>

In some embodiments, the conjugates of the invention comprise an RBD _SA _VAR Polypeptides and polypeptides (S1056-1209, "Spep 4"). In some embodiments, the conjugate comprises a polypeptide having a sequence as set forth in SEQ ID NO:58, and a multi-epitope polypeptide of the amino acid sequence shown in seq id no.

SEQ ID NO：58>

In some embodiments, the conjugates of the invention comprise a polypeptide (N276-411, "Npep 2") and a polypeptide (S125-250, "Spep 1"). In some embodiments, the conjugate comprises a polypeptide having a sequence as set forth in SEQ ID NO:63, and a polypeptide having the amino acid sequence shown in seq id no.

SEQ ID NO：61>

In some embodiments, the heterologous polypeptide is an immunoglobulin domain, particularly the Fc region of an antibody. More particularly, the heterologous polypeptide is the heavy and/or light chain of an antibody.

In some embodiments, the antibody is an IgG antibody, preferably an IgG1 or IgG4 antibody, or even more preferably an IgG4 antibody.

In some embodiments, the antibody is a chimeric antibody, particularly a chimeric mouse/human antibody.

In some embodiments, the antibody is a humanized antibody.

Chimeric or humanized antibodies can be prepared based on the sequences of murine monoclonal antibodies prepared as described above. DNA encoding heavy and light chain immunoglobulins can be obtained from murine hybridomas of interest and engineered to contain non-murine (e.g., human) immunoglobulin sequences using standard molecular biology techniques. For example, to generate chimeric antibodies, the murine variable region can be linked to a human constant region using methods known in the art (see, e.g., U.S. Pat. No. 4,816,567, cabill et al). To generate humanized antibodies, murine CDR regions can be inserted into a human framework using methods known in the art. See, for example, winter, U.S. Pat. No. 5,225,539 and Queen et al, U.S. Pat. No. 5,530,101;5,585,089;5,693,762 and 6,180,370.

In some embodiments, the antibody is a human antibody. In some embodiments, transgenic or transchromosomal mice carrying parts of the human immune system other than the mouse system may be used to identify human antibodies. These transgenic and transchromosomal mice include mice referred to herein as HuMAb mice and KM mice, respectively, and are collectively referred to herein as "human Ig mice". HuMAb(Medarex, inc.) comprises a human immunoglobulin gene minilocus (miniloci) encoding unrearranged human heavy chain (μ and γ) and K light chain immunoglobulin sequences, and a targeting mutation that inactivates endogenous μ and K chain loci (see, e.g., lonberg et al, 1994nature 368 (6474): 856-859). In another embodiment, human antibodies may be produced using mice carrying human immunoglobulin sequences on transgenes and transchromosomes, such as mice carrying human heavy chain transgenes and human light chain transchromosomes. Such mice are referred to herein as "KM mice" and are described in PCT publication WO 02/43478 to Ishida et al.

In some embodiments, the antibody is directed against a surface antigen of an APC, and thus the SARS-CoV-2 polypeptide of the present invention targets the cell to elicit an immune response.

In some embodiments, the antibody is selected from the group consisting of antibodies that specifically bind to: DC immunoreceptor (DCIR), MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11B, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, langerhans's, DECTIN-1, B7-2, IFN-gamma receptor and IL-2 receptor, ICAM-1, fey receptor, LOX-1 and ASPGR. In some embodiments, the antibody is specific for a cell surface marker of a professional APC. Preferably, the antibody is specific for a cell surface marker of DC, such as CD83, CMRF-44 or CMRF-56. The antibody may be specific for another cell surface marker of professional APC, such as B cells or macrophages.

In some embodiments, the antibody is specific for CD40 expressed on DCs, B cells, and other APCs in order to recruit more APCs.

In some embodiments, the CD40 antibody is derived from a 12E12 antibody and comprises:

-a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H, said CDR1H having amino acid sequence GFTFSDYYMY (SEQ ID NO: 8), said CDR2H having amino acid sequence YINSGGGSTYYPDTVKG (SEQ ID NO: 9) and said CDR3H having amino acid sequence RGLPFHAMDY (SEQ ID NO: 10);

-and a light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L, said CDR1L having amino acid sequence SASQGISNYLN (SEQ ID NO: 11), said CDR2L having amino acid sequence ysilh (SEQ ID NO: 12) and said CDR3L having amino acid sequence QQFNKLPPT (SEQ ID NO: 13).

In some embodiments, the CD40 antibody is derived from an 11B6 antibody and comprises:

-a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H, said CDR1H having amino acid sequence GYSFTGYYMH (SEQ ID NO: 14), said CDR2H having amino acid sequence RINPYNGATSYNQNFKD (SEQ ID NO: 15), said CDR3H having amino acid sequence EDYVY (SEQ ID NO: 16), and

a light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L, said CDR1L having amino acid sequence RSSQSLVHSNGNTYLH (SEQ ID NO: 17), said CDR2L having amino acid sequence KVSNRFS (SEQ ID NO: 18) and said CDR3L having amino acid sequence SQSTHVPWT (SEQ ID NO: 19).

In some embodiments, the CD40 antibody is derived from a 12B4 antibody and comprises:

-a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H, said CDR1H having amino acid sequence GYTFTDYVLH (SEQ ID NO: 20), said CDR2H having amino acid sequence YINPYNDGTKYNEKFKG (SEQ ID NO: 21), said CDR3H having amino acid sequence GYPAYSGYAMDY (SEQ ID NO: 22), and

A light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L, said CDR1L having amino acid sequence RASQDISNYLN (SEQ ID NO: 23), said CDR2L having amino acid sequence YTS RRHS (SEQ ID NO: 24) and said CDR3L having amino acid sequence HHGNTLPWT (SEQ ID NO: 25).

In some embodiments, the CD40 antibody is selected from mAb1, mAb2, mAb3, mAb4, mAb5, and mAb6 selected as described in table a.

Table a: CD40 antibodies

SEQ ID NO:26 (amino acid sequence of variable heavy chain region (VH) (v 2) of humanized 11B 6)

EVQLVQSGAEVKKPGASVKISCKASGYSFTGYYMHWVKQAHGQGLEWIGRINPYNGATSYNQNFKDRATLTVDKSTSTAYMELSSLRSEDTAVYYCAREDYVYWGQGTTVTVSSAS

SEQ ID NO:27 (amino acid sequence of variable light chain (VL) Vk (v 2) of humanized 11B6 VL)

DVVMTQSPLSLPVTLGQPASISCRSSQSLVHSNGNTYLHWYQQRPGQSPRLLIYKVSNRFSGVPDRFSGSGSGTDFTLKISRVEAEDVGVYFCSQSTHVPWTFGGGTK

SEQ ID NO:28 (amino acid sequence of variable heavy chain region VH (v 3) of humanized 11B 6)

EVQLVQSGAEVKKPGASVKVSCKASGYSFTGYYMHWVRQAPGQGLEWIGRINPYNGATSYNQNFKDRVTLTVDKSTSTAYMELSSLRSEDTAVYYCAREDYVYWGQGTTVTVSSAS

SEQ ID NO:29 (VH amino acid sequence of mAb3 (12B 4))

EVQLQQSGPELVKPGASVKMSCKASGYTFTDYVLHWVKQKPGQGLEWIGYINPYNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARGYPAYSGYAMDYWGQGTSVTVSSAS

SEQ ID NO:30 (VL amino acid sequence of mAb3 (12B 4))

DIQMTQTTSSLSASLGDRVTISCRASQDISNYLNWYQQKPDGTVKLLIYYTSRLHSGVPSRFSGSGSGTDYSLTISNLEQEDIATYFCHHGNTLPWTFGGGTK

SEQ ID NO:31 (VH amino acid sequence of mAb4 (24A 3 HC))

DVQLQESGPDLVKPSQSLSLTCTVTGYSITSDYSWHWIRQFPGNKLEWMGYIYYSGSTNYNPSLKSRISITRDTSKNQFFLQLNSVTTEDSATYFCARFYYGYSFFDYWGQGTTLTVSSAS

SEQ ID NO:32 (VL amino acid sequence of mAb4 (24A 3 KC))

QIVLTQSPAFMSASPGEKVTMTCSASSSVSYMHWYQQKSGTSPKRWIYDTSKLASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCQQWSSNPLTFGAGTK

SEQ ID NO:33 (VH amino acid sequence of mAb 5)

QVQLVQSGAEVKKPGASVKVSCKASGYTFTGYYMHWVRQAPGQGLEWMGWINPDSGGTNYAQKFQGRVTMTRDTSISTAYMELNRLRSDDTAVYYCARDQPLGYCTNGVCSYFDYWGQGTLVTVSSAS

SEQ ID NO:34 (VL amino acid sequence of mAb 5)

DIQMTQSPSSVSASVGDRVTITCRASQGIYSWLAWYQQKPGKAPNLLIYTASTLQSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQANIFPLTFGGGTK

SEQ ID NO:35 (VH amino acid sequence of mAb6 (12E 12 H3 humanized HC))

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGLPFHAMDYWGQGTLVTVSSAS

SEQ ID NO:36 (VL amino acid sequence of mAb6 (humanized K2 12E 12))

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLpPTFGGGTK

In some embodiments, the CD40 antibody is a CD40 agonist antibody. CD40 agonist antibodies are described in WO2010/009346, WO2010/104747 and WO2010/104749. Other anti-CD 40 agonist antibodies being developed include CP-870, 893, which are all human IgG2 CD40 agonist antibodies developed by Pfizer. It binds CD40 with a KD of 3.48X10-10M, but does not block the binding of CD40L (see, e.g., U.S. Pat. No. 7,338,660), and SGN-40, which is a humanized IgG1 antibody developed by Seattle Genetics from mouse antibody clone S2C6, which was raised using a human bladder cancer cell line as an immunogen. SGN-40 binds CD40 with a KD of 1.0X10-9M and acts by enhancing the interaction between CD40 and CD40L, thus exhibiting a partial agonist effect (Francisco J A, et al, cancer Res,60:3225-31, 2000). Even more particularly, the CD40 agonist antibody is selected from mAb1, mAb2, mAb3, mAb4, mAb5, and mAb6 selected as described in table a.

In some embodiments, the heavy or light chain of a CD40 agonist antibody (i.e., a chain not coupled to or fused to one or more SARS-CoV-2 polypeptides of the invention) is coupled to or fused to the CD40 binding domain of CD 40L.

In some embodiments, the CD40 binding domain of CD40L is fused to the C-terminus of the light or heavy chain of the CD40 agonist antibody, optionally through a linker, preferably a FlexV1 linker as described below.

In some embodiments, the antibodies of the invention consist of CD40 agonist antibodies, wherein the heavy chain of the antibody is fused or conjugated to one or more SARS-CoV-2 polypeptides of the invention and the light chain is conjugated or fused to the CD40 binding domain of CD40L (SEQ ID NO: 4).

In some embodiments, the antibody is specific for Yu Langge hanstin.

In some embodiments, the antibody is derived from antibody 15B10 having ATCC accession No. PTA-9852. In some embodiments, the antibody is derived from antibody 2G3 having ATCC accession No. PTA-9853. In some embodiments, the antibody is derived from antibody 91E7, 37C1 or 4C7 as described in WO 2011032161.

In some embodiments, the anti-langerhans antibody comprises a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H of the 15B10 antibody, and a light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L of the 15B10 antibody.

In some embodiments, the anti-langerhans antibody comprises a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H of the 2G3 antibody, and a light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L of the 2G3 antibody.

In some embodiments, the anti-langerhans antibody comprises a heavy chain comprising the complementarity determining regions CDR1H, CDR H and CDR3H of a 4C7 antibody, and a light chain comprising the complementarity determining regions CDR1L, CDR L and CDR3L of a 4C7 antibody.

In some embodiments, the antibody is selected from, for exampleTable BThe selected mAb7, mAb8, mAb9, mAb10, mAb11 and mAb12.

SEQ ID NO:37 (amino acid sequence of heavy chain variable region (VH) of 15B 10)

SVKMSCKASGYTFTDYVISWVKQRTGQGLEWIGDIYPGSGYSFYNENFKGKATLTADKSSTTAYMQLSSLTSEDSAVYFCA

SEQ ID NO:38 (amino acid sequence of light chain variable region (VL) of 15B 10)

ASISCRSSQSLVHSNGNTYLHWYLQKPGQSPKLLIYKVSNRFSGVPDRFSGSGSGTNFTLEISRVEAEDLGLYFCS

SEQ ID NO:39 (amino acid sequence of heavy chain variable region (VH) of 2G 3)

SSVKMSCKASGYTFTDTVISWVKQRTGQGLEWIGDIYPGSGYSFYNENFKGKATLTADKSSTTAYMQLSSLTSEDSAVYFCA

SEQ ID NO:40 (amino acid sequence of light chain variable region (VL) of 2G 3)

VTLTCRSSTGAVTTSNYANWVQEKPDHLFTGLIGGTNNRVSGVPARFSGSLIGDKAALTITGAQTEDEAIYFCA

SEQ ID NO:41 (amino acid sequence of heavy chain of 4C 7)

QVQLQQSGAELVRPGASVTLSCKASGYTFIDHDMHWVQQTPVYGLEWIGAIDPETGDTGYNQKFKGKAILTADKSSRTAYMELRSLTSEDSAVYYCTIPFYYSNYSPFAYWGQGALVTVSAAKTTAPSVYPLAPVCGGTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPALLQSGLYTLSSSVTVTSNTWPSQTITCNVAHPASSTKVDKKIEPRVPITQNPCPPLKECPPCADLLGGPSVFIFPPKIKDVLMISLSPMVTCVVVDVSEDDPDAQISWFVNNVEVHTAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNRALPSPIEKTISKPRGPVRAPQVYVLPPPAEEMTKKEFSLTCMITGFLPAEIAVDWTSNGRTEQNYKNTATVLDSDGSYFMYSKLRVQKSTWERGSLFACSVVHEGLHNHLTTKTISRSLGKAS

SEQ ID NO:42 (amino acid sequence of light chain of 4C 7)

QIVLSQSPAILSASPGEKVTMTCRASSSVSYMHWYQRKPGSSPKPWIYATSNLASGVPARFSGSGSGTSYSLTISRVEAEDAATYYCQQWSSNPLTFGAGTKLELKRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNSWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

In some embodiments, the heterologous polypeptide is conjugated to one or more SARS-CoV-2 polypeptides of the invention by use of chemical conjugation. Several methods for linking or coupling antibodies to conjugate moieties thereof are known in the art. Examples of types of linkers that have been used to couple moieties to antibodies include, but are not limited to, hydrazones, thioethers, esters, disulfides, and peptide-containing linkers, such as valine-citrulline linkers. A linker may be selected that is, for example, susceptible to cleavage at low pH within the lysosomal compartment, or susceptible to cleavage by a protease, such as a protease preferably expressed in tumor tissue, such as cathepsin (e.g., cathepsin B, C, D). Techniques for coupling polypeptides are well known in the art (see, e.g., arnon et al, "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy," in Monoclonal Antibodies And Cancer Therapy (by Reisfeld et al, alan R.Lists, inc., 1985), "Hellstrom et al," Antibodies For Drug Deliverv, "in Controlled Drug Delivery (by Robinson et al, marcel Deiker, inc. second edition, 1987)," Thorpe, "Antibody CarriersOf Cytotoxic Agents In Cancer Therapy: A Review," in Monoclonal Antibodies'84:Biological And Clinical Applications (by Picchera et al, 1985); "Analysis, results, and Future ProspectiVe of the Therapeutic Use of Radiolabeled Antibody In Cancer Therapy," in Monoclonal Antibodies For Cancer Detection And Therapy (by Baldwin et al, academic Press, 1985); and Thorpe et al, 1982, immunol. Rev.62:119-58; see also, e.g., PCT publication WO 89/12624). Typically, the peptide is covalently linked to a lysine or cysteine residue on the antibody through an N-hydroxysuccinimide ester or maleimide functional group, respectively. The use of Engineered cysteines or coupling methods incorporating unnatural amino acids has been reported to improve the homogeneity of conjugates (Axup, j.y., bajjuri, k.m., ritland, m., hutchins, b.m., kim, c.h., kazane, s.a., halder, r., forsyth, j.s., santidrian, a.f., stafin, k., et al (2012), synthesis of site-specific antibody-858, proc. Natl. Acad. Sci. USA 109,16101-16106, junutula, j.r., flugilla, k.m., graham, r.a., parsons, k.l., ha, e., ab, h., bkta, s., n.g., t., dugger, d.2010-47, and the like, and (2012-47, 47-14. Fan, and the like). Junutula et al (Nat Biotechnol.2008; 26:925-32) developed a cysteine-based site-specific coupling called "THIOMAB" (TDC) that purportedly improved the therapeutic index over conventional coupling methods. For ADCs, conjugation to unnatural amino acids that have been incorporated into the antibodies is also being explored; however, the popularity of this approach has not been established (Axup et al 2012). In particular, it is also contemplated by those skilled in the art that Fc-containing polypeptides are engineered with acyl donor glutamine-containing tags (e.g., gin-containing peptide tags or Q-tags) or with endogenous glutamine that is rendered reactive by polypeptide engineering (e.g., by amino acid deletions, insertions, substitutions, or mutations on the polypeptide). The transglutaminase can then be covalently crosslinked with an amine donor reagent (e.g., a small molecule comprising or linked to a reactive amine) to form a population of stable and homogeneous engineered Fc-containing polypeptide conjugates, wherein the amine donor reagent is site-specifically coupled to the Fc-containing polypeptide (via a tag containing an acyl donor glutamine, or accessible/exposed/reactive endogenous glutamine) (WO 2012059882).

In some embodiments, a heterologous polypeptide (particularly any antibody disclosed herein, particularly any CD40 antibody derived from a 12E12 antibody as defined above) is coupled to one or more SARS-CoV-2 polypeptides of the invention via a dockerin domain or domains that allow non-covalent coupling to an fibronectin fusion protein (as described in US20160031988A1 and US20120039916 A1).

In some embodiments, a heterologous polypeptide (particularly any antibody disclosed herein, particularly any CD40 antibody derived from the 12E12 antibody as defined above) is fused to one or more SARS-CoV-2 polypeptides of the invention to form a fusion protein.

In some embodiments, a heterologous polypeptide (particularly any antibody disclosed herein, particularly any CD40 antibody derived from the 12E12 antibody as defined above) may be fused to the N-terminus or C-terminus of the SARS-CoV-2 polypeptide of the invention.

In some embodiments, the SARS-CoV-2 polypeptide of the invention is fused directly or through a linker to a heterologous polypeptide (in particular any antibody disclosed herein, in particular any CD40 antibody derived from the 12E12 antibody as defined above). As used herein, the term "directly" refers to the fusion of the (first or last) amino acid at the terminus (N or C terminus) of the SARS-CoV-2 polypeptide of the invention to the (first or last) amino acid at the terminus (N or C terminus) of a heterologous polypeptide. Such direct fusion may occur naturally as described below (Vigneron et al, science 2004,PMID 15001714), (Warren et al, science 2006,PMID 16960008), (Berkers et al, J.Immunol.2015a, PMID 26401000), (Berkers et al, J.Immunol.2015b, PMID 26401003), (Delong et al, science 2016,PMID 26912858) (Liepe et al, science 2016,PMID 27846572), (Babon et al, nat.Med.2016, PMID 27798614).

In some embodiments, the linker is selected from FlexV1, f2, f3, or f4 as described below.

QTPTNTISVTPTNNSTPTNNsNPKPNP(flexV1，SEQ ID NO：43)

SSVSPTTSVHPTPTSVPPTPTKSSP(f1，SEQ ID NO：44)

PTSTPADSSTITPTATPTATPTIKG(f2，SEQ ID NO：45)

TVTPTATATPSAIVTTITPTATTKP(f3，SEQ ID NO：46)

TNGSITVAATAPTVTPTVNATPSAA(f4，SEQ ID NO：47)

In some embodiments, the fusion proteins of the invention comprise 2, 3, 4, 5, 6, 7, or 8 SARS-CoV-2 polypeptides of the invention, which can be fused to each other directly or indirectly through a linker. In some embodiments, the fusion proteins of the invention comprise polypeptides (M1-110), polypeptides (M132-222), polypeptides (N78-206), polypeptides (N276-411), polypeptides (S125-250), polypeptides (S280-598), polypeptides (S680-1029), and polypeptides (S1056-1209), which may be fused to each other directly or indirectly through a linker.

In some embodiments, the heterologous polypeptide (particularly any antibody disclosed herein, particularly any CD40 antibody derived from a 12E12 antibody as defined above) is not conjugated to a multi-epitope polypeptide comprising an RBD polypeptide fused to a polypeptide (S125-250, "Spep 1").

In some embodiments, a heterologous polypeptide (particularly any antibody disclosed herein, particularly any CD40 antibody derived from a 12E12 antibody as defined above) is not conjugated to a multi-epitope polypeptide comprising a polypeptide (N276-411 "npep 2") fused to a polypeptide (S1056-1209, "Spep 4").

In some embodiments, the conjugate of the invention (designated "Gen2a" or "CD40.Cov 2") consists of a CD40 antibody (in particular any of the antibodies disclosed herein, in particular any CD40 antibody derived from the 12E12 antibody as defined above), wherein:

fusion of the heavy chain of the antibody with the RDB polypeptide, and

the light chain of the antibody is fused to a multi-epitope polypeptide comprising a polypeptide (N276-411 "Npep 2"), a polypeptide (S125-250, "Spep 1") and a polypeptide (S1056-1209, "Spep 4").

In some embodiments, the light chain of the Gen2a antibody is fused to a multi-epitope polypeptide having the formula Npep2-Spep1-f2-Spep4, wherein f2 represents a linker as described herein. In some embodiments, the light chain of the Gen2a antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

In some embodiments, the Gen2a antibody comprises the amino acid sequence set forth in SEQ ID NO:48 and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:49, and a light chain of the amino acid sequence shown in seq id no.

SEQ ID NO:48> [ H anti-CD 40VH3-LV-hIgG 4H-C-Virus SARS-CoV-2-spike-RBD ]

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAFDTAVYYCARRGLPFHAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKASRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNF

SEQ ID NO:49> [ h anti-CD 40VK2-LV-hIgGK-C-f 4-Virus SARS-CoV-2-Npep2-Spep1-f2-Spep4 ]

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECASTNGSITVAATAPTVTPTVNATPSAAASRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMARNVVIKVCEFQFSNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTRPTSTPADSSTITPTATPTATPTIKGAPHGWFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFvSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQY

In some embodiments, the conjugate of the invention (designated or "Gen2 b") consists of a CD40 antibody (in particular any antibody disclosed herein, in particular any CD40 antibody derived from a 12E12 antibody as defined above), wherein:

Heavy chain of antibody fusion with a polyepitope polypeptide comprising polypeptide (S125-250, "Spep 1") and polypeptide (S1056-1209, "Spep 4")

The light chain of the antibody is fused to a multi-epitope polypeptide comprising a polypeptide (N276-411, "Npep 2") and an RBD polypeptide.

In some embodiments, the heavy chain of the Gen2b antibody is fused to a multi-epitope polypeptide having the formula Spep1-f2-Spep4, wherein f2 represents a linker as described herein. In some embodiments, the heavy chain of the Gen2b antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:6, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

In some embodiments, the light chain of Gen2b is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:7, and a polypeptide having the amino acid sequence shown in FIG. 7

In some embodiments, the Gen2b antibody comprises the amino acid sequence set forth in SEQ ID NO:50 and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:51, and a light chain of the amino acid sequence shown in seq id no.

SEQ ID NO:50> [ H anti-CD 40VH3-LV-hIgG4H-C-Flex-v 1-Virus SARS-CoV-2-Spep1-f2-pep4]

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGLPFHAMDYWGQGTLVTVSSASTKGPSVFPIAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAKLQSSGLYSLSSVVTVPsSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPFVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKASQTPTNTISVTPTNNSTPTNNSNPKPNPARNVVIKVCEFQFSNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTRPTSTPADSSTITPTATPTATPTIKGAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYFPQIITTDNTFVSGNCDVVIGTVNNTVVDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQY

SEQ ID NO:51> [ h anti-CD 40VK2-LV-hIgGK-C-f 4-Virus SARS-CoV-2-Npep 2-spike-RBD ]

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECASTNGSITVAATAPTVTPTVNATPSAAASRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQEAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMASRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNF

In some embodiments, the conjugate of the invention (designated "Gen2 c") consists of a CD40 antibody (in particular any antibody disclosed herein, in particular any CD40 antibody derived from a 12E12 antibody as defined above), wherein:

The heavy chain of the antibody is fused to a multi-epitope polypeptide comprising a polypeptide (N276-411 "Npep 2"), a polypeptide (S125-250, "Spep 1") and a polypeptide (S1056-1209, "Spep 4"), and

antibody light chain and RBD polypeptide fusion

In some embodiments, the heavy chain of the Gen2c antibody is fused to a multi-epitope polypeptide having the formula Npep2-Spep1-f2-Spep4, wherein f2 represents a linker as described herein. In some embodiments, the heavy chain of the Gen2c antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

In some embodiments, the Gen2c antibody comprises the amino acid sequence set forth in SEQ ID NO:52 and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:53, and a light chain of the amino acid sequence shown in seq id no.

SEQ ID NO:52> [ H anti-CD 40VH3-LV-hIgG4H-C-f 4-Virus SARS-CoV-2-Npep2-Spep1-f2-Spep4]

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGLPFHAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKASTNGSITVAATAPTVTPTVNATPSAAASRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKkADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMARNVVIKVCEFQFSNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLFGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTRPTSTPADSSTITPTATPTATPTIKGAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQY

SEQ ID NO:53> [ h anti-CD 40VK 2-LV-hIgGK-C-Virus SARS-CoV-2-spike-RBD ]

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECASRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGKIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVEGFNCYFPLQSYGFQPTNGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNF

In some embodiments, the conjugate of the invention (designated "CD 40.cov2v") consists of a CD40 antibody (in particular any antibody disclosed herein, in particular any CD40 antibody derived from a 12E12 antibody as defined above), wherein:

fusion of the heavy chain of the antibody with the RBD polypeptide, and

the light chain of the antibody is fused to a multi-epitope polypeptide comprising a polypeptide (N276-411 "Npep 2"), a polypeptide (S125-250, "Spep 1"), and a polypeptide (S1056-1209, "Spep 4").

In some embodiments, the light chain of the cd40.cov2v antibody is fused to a multi-epitope polypeptide having the formula Npep2-Spep1-f2-Spep4, wherein f2 represents a linker as described below. In some embodiments, the light chain of the cd40.cov2v antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

In some embodiments, the cd40.cov2v antibody comprises the amino acid sequence as set forth in SEQ ID NO:54 and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:55, and a light chain of the amino acid sequence shown in seq id no.

SEQ ID NO:54> [ H anti-CD 40VH 3-LV-hIgGK-C-virus SARS-CoV-2-spike-RBDC 221S variant (south Africa mutation 20H/501Y V2) ]

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGLPFHAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKASRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGNIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVKGFNCYFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNF

SEQ ID NO:55> [ h anti-CD 40VK2-LV-hIgGK-C-f 4-Virus SARS-Cov-2-Npep2-Spep1C12S-f2-Spep4]

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECASTNGSITVAATAPTVTPTVNATPSAAASRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMARNVVIKVCEFQFSNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLEGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYLTRPTSTPADSSTITPTATPTATPTIKGAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFvSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQY

In some embodiments, the conjugate of the invention (designated "CD40.N2. Rbdv") consists of a CD40 antibody (in particular any CD40 antibody derived from a 12E12 antibody as defined above), wherein:

the heavy chain of the antibody is fused to a polypeptide (N276-411 "Npep 2"), and

the light chain of the antibody is fused to the RBD polypeptide,

in some embodiments, the cd40.N2.Rbdv comprises the amino acid sequence as set forth in SEQ ID NO:56 and a heavy chain having the amino acid sequence as set forth in SEQ ID NO: 57.

SEQ ID NO:56> [ H anti-CD 40VH3-LV-hIgG4H-C-f 4-Virus SARS-Cov-2-Npep2]

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAFDTAVYYCARRGLPFHAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKASTNGSITVAATAPTVTPTVNATPSAAASRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSM

SEQ ID NO：57>[ h anti-CD 40VK 2-LV-hIgGK-C-virus SARS-CoV-2-spike-RBD _C221 sSA var]

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSISSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECASRVQPTESIVRFPNTTNLCPFGEVFNATRFASVYAWNRKRTSNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGNIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVKGFNCYFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNF

In some embodiments, the conjugates of the invention consist of a CD40 antibody (designated "CD 40.n2.rbdv-2") (in particular any CD40 antibody derived from a 12E12 antibody as defined above), wherein:

fusion of the heavy chain of the antibody with the RBD polypeptide, and

the light chain of the antibody is fused to a polypeptide (N276-411 "Npep 2").

In some embodiments, the conjugate of the invention (designated "CD40.Rbdv s4. N2") consists of a CD40 antibody (in particular any CD40 antibody derived from a 12E12 antibody as defined above), wherein:

the heavy chain of the antibody is fused to a multi-epitope polypeptide comprising an RBD polypeptide and a polypeptide (S1056-1209, "Spep 4"), and

the light chain of the antibody is fused to a polypeptide (N276-411 "Npep 2").

In some embodiments, the heavy chain of the cd40 rbdv s4 n2 antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:58, and a multi-epitope polypeptide of the amino acid sequence shown in seq id no.

In some embodiments, cd40.Rbdv s4.N2 comprises the amino acid sequence as set forth in SEQ ID NO:59 and a heavy chain having the amino acid sequence shown in SEQ ID NO:60, and a light chain of the amino acid sequence shown in seq id no.

SEQ ID NO：59>[ H anti-CD 40VH3-LV-hIgG 4H-C-Virus SARS-CoV-2-RBD _C221S SA var-Spep4]

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYCARRGLPFHAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKASRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRTSNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVTRGDEVRQTAPGQTGNTADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVKGFNCYFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNFASAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPREGVFVSNGTHWFVTQRNFYEPQIITTDNTPVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQY

SEQ ID NO:60> [ h anti-CD 40VK2-LV-hIgGK-C-f 4-Virus SARS-CoV-2-Npep2]

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKsFNRGEcASTNGSITVAATAPTVTPTVNATPSAAASRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQEAPSASAFFGMSRIGMEVTPsGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSM

In some embodiments, the conjugate of the invention (designated "CD 40.n2s1.rbdvs4") consists of a CD40 antibody (in particular any CD40 antibody derived from the 12E12 antibody as defined above), wherein:

the heavy chain of the antibody is fused to a multi-epitope polypeptide comprising a polypeptide (N276-411 "Npep 2") and a polypeptide (S125-250, "Spep 1"),

the light chain of the antibody is fused to a multi-epitope polypeptide comprising an RBD polypeptide and a polypeptide (S1056-1209, "Spep 4").

In some embodiments, the light chain of the cd40.n2s1.rbdvs4 antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:58, and a multi-epitope polypeptide of the amino acid sequence shown in seq id no. In some embodiments, the heavy chain of the cd40.n2s1.rbdvs4 antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:61, and a polypeptide having the amino acid sequence shown in seq id no.

In some embodiments, the cd40.n2s1.rbdvs4 antibody comprises the amino acid sequence as set forth in SEQ ID NO:62 and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:63, and a light chain of the amino acid sequence shown in seq id no.

SEQ ID NO：62>[ H anti-CD 40VH3-LV-hIgG4H-C-f 4-Virus SARS-CoV-2-Npep2-Spep1 _C12S ]

EVQLVESGGGLVQPGGSLKLSCATSGFTFSDYYMYWVRQAPGKGLEWVAYINSGGGSTYYPDTVKGRFTISRDNAKNTLYLQMNSLRAEDTAVYYcARRGLPFHAMDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFEGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVFHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGKASTNGSITVAATAPTVTPTVNATPSAAASRRGPEQTQGNFGDQELIRQGTDYKHWPQIAQFAPSASAFFGMSRIGMEVTPSGTWLTYTGAIKLDDKDPNFKDQVILLNKHIDAYKTFPPTEPKKDKKKKADETQALPQRQKKQQTVTLLPAADLDDFSKQLQQSMASNVVIKVCEFQFSNDPFLGVYYHKNNKSWMESEFRVYSSANNCTFEYVSQPFLMDLFGKQGNFKNLREFVFKNIDGYFKIYSKHTPINLVRDLPQGFSALEPLVDLPIGINITRFQTLLALHRSYL

SEQ ID NO：63>[ h anti-CD 40VK 2-LV-hIgGK-C-virus SARS-CoV-2-RBD _C221S SA var-Spep4]

DIQMTQSPSSLSASVGDRVTITCSASQGISNYLNWYQQKPGKAVKLLIYYTSILHSGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCQQFNKLPPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECASRVQPTESIVRFPNITNLCPFGEVFNATRFASVYAWNRKRISNCVADYSVLYNSASFSTFKCYGVSPTKLNDLCFTNVYADSFVIRGDEVRQIAPGQTGNIADYNYKLPDDFTGCVIAWNSNNLDSKVGGNYNYLYRLFRKSNLKPFERDISTEIYQAGSTPCNGVKGFNCYFPLQSYGFQPTYGVGYQPYRVVVLSFELLHAPATVCGPKKSTNLVKNKSVNFASAPHGVVFLHVTYVPAQEKNFTTAPAICHDGKAHFPRFGVFVSNGTHWFVTQRNFYEPQIITTDNTFVSGNCDVVIGIVNNTVYDPLQPELDSFKEELDKYFKNHTSPDVDLGDISGINASVVNIQKEIDRLNEVAKNLNESLIDLQELGKYEQY

In some embodiments, the RBD polypeptides comprised in the conjugates Gen2a, gen2b, gen2c, cd40.Cov2v, cd40.N2.Rbdv-2, cd40.Rbdvs4.N2 or cd40.N2s1.Rbdvs4 as defined above consist of the following amino acids: from SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a non-naturally occurring C538S mutation and at least one naturally occurring mutation, particularly in SEQ ID NO: one or more of bits 417, 452, 484, or 501 of 3.

In particular, the RBD polypeptides comprised in the conjugates Gen2a, gen2b, gen2c, cd40.cov2v, cd40.n2.rbdv-2, cd40.rbdvs4.n2 or cd40.n2s1.rbdvs4 as defined above may consist of:

● In SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and includes naturally occurring mutations in N501Y and non-naturally occurring mutations in C538S;

● In SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a naturally occurring mutation of K417T, E484K, N Y and a non-naturally occurring mutation of C538S;

● In SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a naturally occurring mutation of K417N, E484K, N501Y and a non-naturally occurring C538S mutation ("RBD) _{South Africa variants} ""; or alternatively

● In SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 in 3Amino acids ("RBD") comprising both E484Q, L452N naturally occurring mutations and non-naturally occurring C538S mutations _{Indian variants} ”)。

The RBD polypeptides comprised in the conjugates Gen2a, gen2b, gen2c, cd40.cov2v, cd40.n2.rbdv-2, cd40.rbdvs4.n2 or cd40.n2s1.rbdvs4 as defined above may preferably consist of: SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a naturally occurring mutation of K417N, E484K, N501Y and a non-naturally occurring C538S mutation ("RBD) _{South Africa variants} ”)。

Methods of producing the polypeptides of the invention:

the SARS-CoV-2 polypeptides, fusion proteins and antibodies of the invention can be produced by any technique known in the art, such as, but not limited to, any chemical, biological, genetic or enzymatic technique (alone or in combination). Knowing the amino acid sequence of the desired sequence, one skilled in the art can readily produce the polypeptide by standard techniques for producing polypeptides. For example, it may be synthesized using well known solid phase methods, preferably using commercially available peptide synthesis equipment (e.g., applied Biosystems, equipment manufactured by Foster City, california) and following manufacturer's instructions. Alternatively, the polypeptides and fusion proteins of the invention may be synthesized by recombinant DNA techniques now well known in the art. For example, after incorporating a DNA sequence encoding the desired (poly) peptide into an expression vector, such vector is introduced into a suitable eukaryotic or prokaryotic host in which the desired polypeptide will be expressed, these fragments may be obtained as DNA expression products, after which they may be isolated from the host using well-known techniques.

For recombinant expression, the DNA construct preferably comprises sequences encoding signal peptides in the N-terminus of the heavy and light chains of the conjugate according to the invention.

Pharmaceutical and vaccine compositions:

SARS-CoV-2 polypeptides, and conjugates comprising fusion proteins and antibodies, as described herein, and antibodies can be administered as part of one or more pharmaceutical compositions. Unless any conventional carrier medium is incompatible with the polypeptides of the invention, such as by producing any undesirable biological effect or interacting in a deleterious manner with any of the other components of the pharmaceutical composition, its use is contemplated as being within the scope of the invention. Some examples of materials that may be used as pharmaceutically acceptable carriers include, but are not limited to, sugars such as lactose, glucose, and sucrose; starches, such as corn starch, potato starch; cellulose and its derivatives such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; powdery tragacanth; malt; gelatin; talc; excipients, such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil; safflower oil and sesame oil; olive oil; corn oil and soybean oil; ethylene glycol; such as propylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffering agents such as magnesium hydroxide, aluminum hydroxide; alginic acid; non-thermal raw water; isotonic saline; ringer's solution; ethanol and phosphate buffer solutions, as well as other non-toxic compatible lubricants (e.g., sodium lauryl sulfate and magnesium stearate), as well as coloring agents, releasing agents, coating agents, sweetening, flavoring and perfuming agents, preserving and antioxidant agents, can also be present in the composition (at the discretion of the formulator).

SARS-CoV-2 polypeptides, as described herein, as well as conjugates comprising fusion proteins and antibodies, are particularly useful in the preparation of vaccine compositions.

In some embodiments, the vaccine compositions of the invention comprise one or more SARS-CoV-2 polypeptides as antigens. One of the advantages of the SARS-CoV-2 polypeptide of the invention is that it comprises multiple epitopes, including HLA class I restriction epitopes and/or HLA class II restriction epitopes and/or antibody epitopes.

In some embodiments, the vaccine composition of the invention comprises 2, 3, 4, 5, 6, 7, or 8 SARS-CoV-2 polypeptides of the invention. In some embodiments, the vaccine compositions of the present invention desirably comprise polypeptide (M1-110), polypeptide (M132-222), polypeptide (N78-206), polypeptide (N276-411), polypeptide (S125-250), polypeptide (S280-598), polypeptide (S680-1029), and polypeptide (S1056-1209).

In some embodiments, the vaccine compositions of the invention comprise one or more conjugates of the invention (including fusion proteins and antibodies as described herein) as an antigen.

Thus, in some embodiments, the vaccine compositions of the present invention comprise an adjuvant. In some embodiments, the adjuvant is alum. In some embodiments, the adjuvant is Incomplete Freund's Adjuvant (IFA) or other oil-based adjuvant, which is present in a weight ratio (w/w) of 30-70%, preferably 40-60%, more preferably 45-55%. In some embodiments, the vaccine compositions of the invention comprise at least one Toll-like receptor (TLR) agonist selected from the group consisting of TLR1, TLR2, TLR3, TLR4, TLR5, TLR6, TLR7 and TLR8 agonists.

Polynucleotides and uses thereof:

another object of the present invention relates to polynucleotides encoding SARS-CoV-2 polypeptides or conjugates of the invention (including fusion proteins and antibodies of the invention). The polynucleotide preferably comprises sequences encoding signal peptides in the N-terminal ends of the heavy and light chains of the conjugates according to the invention, such that the conjugates can be expressed and secreted in vivo.

Typically, the polynucleotide is a DNA or RNA molecule, which may be contained in any suitable vector, such as a plasmid, cosmid, episome, artificial chromosome, phage, or viral vector.

The polynucleotides of the invention are particularly useful for vaccinating a subject in need thereof.

There are a variety of methods for delivering a polynucleotide to a subject as defined herein. For example, polynucleotides of the invention may be formulated with cationic polymers, including cationic liposomes. Other liposomes also represent an effective means of formulating and delivering self-acidic nucleic acid molecules. Alternatively, the DNA may be incorporated into a viral vector, viral particle or bacteria for pharmacological delivery. Viral vectors may be infectious, attenuated (with mutations that reduce the ability to induce disease) or replication defective. Methods of using DNA to prevent deposition, accumulation or activity of pathogenic self-proteins can be enhanced by using viral vectors, or other delivery systems that increase the humoral response to the encoded self-antigen. In some embodiments, the DNA may be coupled to a solid support, including gold particles, polysaccharide-based supports, or may be coupled to other particles or beads that may be delivered by injection, inhalation, or by particle bombardment (ballistic delivery). Methods of delivering nucleic acid formulations are known in the art. See, for example, U.S. Pat. nos. 5,399,346, 5,580,859, and 5,589,466. Many virus-based systems have been developed for transfer into mammalian cells. For example, a retroviral system has been described (U.S. patent No. 5,219,740; miller et al, biotechniques 7:980-990 (1989); miller, human Gene Therapy:5-14, (1990); scarpa et al, virology 180:849-852 (1991), burns et al, proc. Natl Acad. Sci. USA 90:8033-8037 (1993), and Boris-Lawrei and Temin, curr. Opin. Genet. Development. 3:102-109 (1993) also describe many adenovirus vectors, see, e.g., haj-Ahmad et al, J. Virol.57:267-274 (1986), bett et al, J. Virol.67:5911-5921 (1993), mitterer et al, human Gene Therapy:717-729 (1994), seris et al, virol.68:933-940 (1994), barr et al, gene Therapy 1:51-58 (1994), berkner, bioniques 6:616-629 (1988), and 92-Ahma et al, J. No. 35:92-37, J. 6, and Virol. 35:37-37-95 (1986), AAV 37-37, 6, and Virol. 35:35, 6:37-37, and Virol. 35, 6:35, and Virol. 35, see, lee.g., lee.35:37-37-95, and Lee.g., lee.35, lee.g., lee.35, no. 6, lee.35, and Lee.g., see, no. 6, lee.35, and WO-7, lee.35, and WO-7, lee.35, lee.J. 35, lee.J. J. J. J. 7, A. J. J. Lee.J. Lee J. Leg., viter, A. 7, A. J. Vav. 7, lev. 7, E7, EK 6, EK 6, EK Abs. Viv AgK, EK AGApp Viv AgK AAbApp Viv AcApp AcApto AcIEAbIEAbIEAbIEAbIEAbIEAbIAAcApto AcApto AcApL AcApto AcAcApIEAbAcApIEAbAcApto AcAcApto AcAcAcAcAcAcAcAcAcAcAcAcAcAcAcAcAcAcAcAcAAcAAcAcAcAAcAAcAcAcAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcAAcViViViViViViViViViViViViVi, exp. Med.179:1867-1875 (1994).

In some embodiments, the polynucleotides of the invention are delivered in the absence of a viral vector. For example, the polynucleotide may be packaged into a liposome prior to delivery to a subject. Lipid encapsulation is typically accomplished using liposomes that are capable of stably binding or entrapping and retaining nucleic acids. For reviews on the use of liposomes as nucleic acid delivery vehicles, see, e.g., hug et al, biochim. Biophys acta.1097:1-17 (1991); straubinger et al, in Methods of Enzymology,101:512-527 (1983). Alternatively, the polynucleotide is delivered by electroporation (i.e., muscle delivery by electroporation).

In some embodiments, the polynucleotide is delivered by intramuscular ("IM") injection. In some embodiments, the acidic nucleic acid molecules of the invention are delivered by intranasal, oral, subcutaneous, intradermal, intravenous, mucosal, by skin imprinting, or by particles attached to or delivered to or through the dermis. Alternatively, the polynucleotide may be delivered to skin cells by topical application with or without liposomes or charged lipids. Yet another option is to deliver the nucleic acid as an inhalant.

The treatment method comprises the following steps:

SARS-CoV-2 polypeptides, and conjugates comprising fusion proteins and antibodies, as described herein, and pharmaceutical or vaccine compositions are particularly useful for inducing an immune response against SARS-CoV-2 and thus may be used for vaccine purposes.

Accordingly, another object of the present invention relates to a method of vaccinating a subject in need thereof against SARS-CoV-2 comprising administering a therapeutically effective amount of one or more SARS-CoV-2 polypeptides of the invention.

Another object of the invention relates to a method of vaccinating a subject in need thereof against SARS-CoV-2 comprising administering a therapeutically effective amount of one or more conjugates as described herein.

Another object of the invention relates to a method of vaccinating a subject in need thereof against SARS-CoV-2 comprising administering a therapeutically effective amount of a pharmaceutical or vaccine composition as described herein.

Another object of the invention relates to a method of vaccinating a subject in need thereof against SARS-CoV-2 comprising administering a therapeutically effective amount of a polynucleotide as described herein.

In some embodiments, SARS-CoV-2 polypeptides, and conjugates comprising fusion proteins and antibodies, as described herein, are particularly useful in the treatment of Covid-19 as a pharmaceutical or vaccine composition.

In some embodiments, the subject may be a human or any other animal susceptible to coronavirus infection (e.g., birds and mammals) (e.g., domestic animals such as cats and dogs; livestock and farm animals such as horses, cattle, pigs, chickens, etc.). Typically, the subject is a mammal, including a non-primate (e.g., camel, donkey, zebra, cow, pig, horse, goat, sheep, cat, dog, rat, and mouse) and a primate (e.g., monkey, chimpanzee, and human). In some embodiments, the subject is a non-human animal. In some embodiments, the subject is a farm animal or a pet. In some embodiments, the subject is a human. In some embodiments, the subject is a human infant. In some embodiments, the subject is a human child. In some embodiments, the subject is an adult. In some embodiments, the subject is an elderly person. In some embodiments, the subject is a premature infant.

In some embodiments, the subject may be symptomatic or asymptomatic.

Typically, the active ingredients of the present invention (i.e., SARS-CoV-2 polypeptides, and conjugates comprising fusion proteins and antibodies, as well as pharmaceutical or vaccine compositions, as described herein) are administered to a subject in a therapeutically effective amount. It will be appreciated that the total daily amount of the compounds and compositions of the present invention will be determined by the attending physician within the scope of sound medical judgment. The specific therapeutically effective dose level for any particular subject will depend on a variety of factors, including the disorder to be treated and the severity of the disorder; the activity of the particular compound employed; the specific composition employed, the age, weight, general health, sex and diet of the subject; the time of administration, route of administration and rate of excretion of the particular compound employed; duration of treatment; a medicament for use in combination with or simultaneously with the particular polypeptide employed; and similar factors well known in the medical arts. For example, it is well within the skill in the art to begin administering the compound at a level below that required to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved. However, the daily dosage of the product may vary from 0.01 to 1,000mg per adult per day. In particular, the composition comprises 0.01, 0.05, 0.1, 0.5, 1.0, 2.5, 5.0, 10.0, 15.0, 25.0, 50.0, 100, 250 and 500mg of active ingredient for symptomatic adjustment of the dose to the subject to be treated. The medicament generally comprises from about 0.01mg to about 500mg of the active ingredient, in particular from 1mg to about 100mg of the active ingredient. An effective amount of the drug is typically provided at a dosage level of 0.0002mg/kg to about 20mg/kg body weight per day, particularly about 0.001mg/kg to 7mg/kg body weight per day.

SARS-CoV-2 polypeptides, and conjugates comprising fusion proteins and antibodies, as well as pharmaceutical or vaccine compositions, as described herein, may be administered to a subject by any route of administration, particularly by oral, nasal, rectal, topical, buccal (e.g., sublingual), parenteral (e.g., subcutaneous, intramuscular, intradermal, or intravenous) and transdermal administration, although the most suitable route in any given case will depend on the nature and severity of the condition being treated, as well as the nature of the particular active agent being used.

In some embodiments, SARS-CoV-2 polypeptides, and conjugates comprising fusion proteins and antibodies, as well as pharmaceutical or vaccine compositions, as described herein, can be administered to a subject in combination with any known therapeutic agent or method, e.g., for vaccination against SARS-CoV-2 coronavirus. Non-limiting examples of such known treatments include, but are not limited to, antiviral agents such as ritonavir, lopinavir, ritonavir, hydroxychloroquine, and chloroquine. In some embodiments, SARS-CoV-2 polypeptides, and conjugates comprising fusion proteins and antibodies, as well as pharmaceutical or vaccine compositions, as described herein, are administered in combination with an immune checkpoint inhibitor. Examples of immune checkpoint inhibitors include PD-1 antagonists, PD-L2 antagonists, CTLA-4 antagonists, VISTA antagonists, TIM-3 antagonists, LAG-3 antagonists, IDO antagonists, KIR2D antagonists, A2AR antagonists, B7-H3 antagonists, B7-H4 antagonists and BTLA antagonists. In some embodiments, a PD-1 (Programmed Death) -1) axis antagonist Including PD-1 antagonists (e.g., anti-PD-1 antibodies), PD-L1 (programmed death ligand-1) antagonists (e.g., anti-PD-L1 antibodies), and PD-L2 (programmed death ligand-2) antagonists (e.g., anti-PD-L2 antibodies). In some embodiments, the anti-PD-1 antibody is selected from MDX-1106 (also known as Nivolumab, MDX-1106-04, ONO-4538, BMS-936558 and) Merck 3475 (also known as Pembrolizumab), MK-3475, lanbolizumab (Lambrolizumab), and @>And SCH-900475) and CT-011 (also known as Pidilizumab), hBAT and hBAT-1. In some embodiments, the PD-1 binding antagonist is AMP-224 (also known as B7-DCIg). In some embodiments, the anti-PD-L1 antibody is selected from the group consisting of YW243.55.S70, MPDL3280A, MDX-1105 and MEDI4736.MDX-1105 (also called BMS-936559) is an anti-PD-L1 antibody described in WO 2007/005874. Antibody yw243.55.s70 is anti-PD-L1 described in WO 2010/077634 A1. MEDI4736 is an anti-PD-L1 antibody described in WO2011/066389 and US2013/034559.MDX-1106 (also known as MDX-1106-04, ONO-4538 or BMS-936558) is an anti-PD-1 antibody, described in U.S. Pat. No. 8,008,449 and WO2006/121168.Merck 3745 (also known as MK-3475 or SCH-900475) is an anti-PD-1 antibody, described in U.S. Pat. No. 8,345,509 and WO2009/114335.CT-011 (Pidazumab) (also known as hBAT or hBAT-1) is an anti-PD-1 antibody, described in WO2009/101611.AMP-224 (also known as B7-DCIg) is a PD-L2-Fc fusion soluble receptor described in WO2010/027827 and WO2011/066342. Alemtuzumab (Atezolimumab) is an anti-PD-L1 antibody, described in U.S. patent No. 8,217,149. A Wei Shankang (Avelumab) is an anti-PD-L1 antibody, described in US 20140341917.CA-170 is WO2015033301 &PD-1 antagonists described in WO 2015033299. Other anti-PD-1 antibodies are disclosed in U.S. patent No. 8,609,089, US 2010028330 and/or US 20120114649. In some embodiments, the PD-1 inhibitor is an anti-PD-1 antibody selected from Nivolumab, pembrolizumab, or pilizumab. In some embodiments, the PD-L1 antagonist is selected from the group consisting of avilamab, BMS-936559,CA-170, devalumab (Durvalumab), MCLA-145, SP142, STI-A1011, STIA1012, STI-A1010, STI-A1014, A110, KY1003 and alemtuzumab, preferably Al Wei Shankang, devaluzumab or alemtuzumab.

The invention will be further illustrated by the following figures and examples. However, these examples and drawings should not be construed as limiting the scope of the invention in any way.

Drawings

FIG. 1 design and physical properties of anti-CD 40.SARS-CoV-2Gen2a, 2b, 2c fusion proteins. A) Schematic of SARS-CoV-2Gen2a, 2b, 2c, CD40.CoV2v, CD40.N2.RBDv-2, CD40.RBDvS4.N2, CD40.N2S1.RBDvS4 fusion proteins. B) 3 different batches of protein A purified anti-CD 40.SARS-CoV-2Gen2a, 2b, 2c fusion proteins were analyzed by reducing and non-reducing SDS PAGE stained with Coomassie Brilliant blue R250. In both gels, the molecular weight markers are shown in the rightmost lane (BenchMark pre-stained protein ladder; 180, 115, 82, 64, 49, 37, 26, 19, 15, 6kDa; from thermosfisher).

FIG. 2 binding of anti-CD 40 Gen2a, gen2B and Gen2c fusion proteins to A) human CD40 and B) human IgG. Fusion proteins corresponding to the indicated constructs were normalized by molar concentration and serial dilutions were added to the surface coated with human CD40 extracellular domain protein (top panel), or goat anti-human IgG polyclonal serum, incubated to allow binding, washed, then incubated with horseradish peroxidase (HRP) conjugated anti-human IgG reagent, followed by washing and development with HRP chromogenic substrate.

FIG. 3 test of CD 40-targeting SARS-CoV2pep antigen in Gen2a, gen2b and Gen2c fusion proteins by in vitro expansion of SARS-CoV-2 specific T cells in a convalescence donor PBMC culture of SARS-CoV-2 infection. SARS-CoV-2 ⁺ 1PBMC of donor patient were incubated with IL-2 and anti-CD 40 Gen2a, gen2b, gen2c fusion proteins (1 nM or 10 nM) for 9 days, then stimulated with peptide libraries specific for SARS-CoV-2N and S regions for 6 hours by Brefeldin-A, then analyzed by Intracellular Cytokine Staining (ICS). 1 = DMSO stimulation; 2. 3, 4 = stimulation with a peptide pool comprising N regions; 5. 6, 7 = packStimulating the peptide library containing the RBD region; 8 = SEB polyclonal stimulation; 9. 10 = stimulation with a peptide pool comprising Spep1 region; 11. 12 = stimulation with a peptide pool comprising Spep4 region. Bar graphs show CD4 with intracellular interferon gamma (i+), intracellular tumor necrosis factor alpha (t+), or both (i+t+) ⁺ Percent stacking value of T cells.

Fig. 4: examples of relative expression of various H-chain and L-chain construct cotransfection experiments were assessed by CD40 binding ELISA. ELISA plates were coated with 1ug/ml human CD40 extracellular domain protein and bound antibody was detected by anti-human IgG-HRP reagent. CHO-S transfection supernatant was not diluted at the beginning (at the point of 1ug/ml for control anti-human CD40 12e12 higgg 4 antibody batch, PAB 2220). Construct nomenclature is detailed in table 2.

Fig. 5: multifunctional cd4+ T cell response induced by the anti-cd40.gen2 vaccine in convalescent patients with covd-19. For PBMCs from 13 covd patients, stimulation was performed on day 0 with 1nM of the anti-cd 40.gen2a vaccine, or equimolar amounts of peptide pool overlapping each antigen (RBD, S1, S4, N2) contained in the vaccine. After 8 days of incubation with IL-2, cells were re-stimulated or not stimulated (unstimul.) using each individual peptide pool (1 ug/ml). In vitro T cell expansion was analyzed by flow cytometry. Single CD4 ⁺ Mean of T cell responses (sum of IFNγ, TNF. Alpha. And IL-2) and PBMC stimulated with vaccine. Multiple comparison one-way ANOVA assays were used for statistical analysis. (ns, no significance; p)<0,05；***p<0,001；****p<0,0001)。

Fig. 6: human B cell frequency of SARS-CoV-2S protein specific IgG was converted in hu-B cells of the spleen of hu-mice 44 days after the initial injection. A. A schematic overview of vaccination strategies in NSG humanized (hu) mice, comprising four experimental groups of 7 to 9 animals each. B. First, hu mouse spleen cells 44 days after immunization were incubated with PE-SARS-CoV-2S protein at 4℃for 1 hour. After the washing step, the cells were stained with anti-mouse CD45, anti-human (h) CD45, anti-hCD 19, anti-hCD 20, anti-hIgG at 4 ℃ for 30 min. Staining of spleen cells also included an active marker (livehead aqua). Cells were washed twice with FACS buffer (PBS 1% FCS) and collected on LSRII flow cytometry (BD Biosciences). Analysis was performed on FlowJo v.10.7.1.

Example 1:

undeniably, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has become the greatest global health threat faced by humans in this century. The speed, rate, and observed increase in mortality of global spread present challenges to public health, socioeconomic and scientific. SARS-CoV-2 can cause respiratory syndrome with clinical pathology similar to mild upper respiratory disease (common cold-like symptoms), and occasionally severe lower respiratory disease and extrapulmonary manifestations, leading to multiple organ failure and death.

There are no therapeutic methods or vaccines available. However, if the virus is stable in the heel of a human population, the SARS-CoV-2 vaccine is critical to reduce morbidity and mortality. In order to accelerate the development of candidate vaccines, it is critical to use a custom and adapted ready-to-use vaccine platform for emerging and re-emerging pathogens. We have established a candidate vaccine against SARS-CoV-2 in a tailored and adaptive DC-targeting vaccine platform against emerging and re-emerging pathogens.

We developed a platform for tailored vaccines that use specific humanized monoclonal antibodies (mabs) to specific DC endocytic receptors to deliver the antigen of interest (common, dominant or cross-reactive) to endogenous DCs. We aim to facilitate antigen presentation and activate antigen-specific immune responses, Even at low antigen doses, strong priming is initiated Is a humoral and cellular immune response of (a). We have generated a large amount of in vitro and in vivo data such thatI of vaccine targeting HIV or HPV Phase study enters clinical development stage[1-6]. We have previously demonstrated that anti-CD 40 mAbs fused to a fragment of HIV T cell epitope are in human [3 ]]、NHP[2]Or humanized mice [1 ]]A multi-epitope T cell response. In addition, anti-CD 40 vaccine fused to HIV.env gp140 in combination with Pox vector, or as enhancer of DNA or DREP HIV vaccine, elicits strong and robust T-cell and B-cell responses [6 ]](P Liljestrom/Y Levy, personal data). The DC-targeting plat formThe bench has now been validated in preclinical stages in a variety of infectious diseases (ebola, influenza, chlamydia, tuberculosis infection) and will advance to phase I clinical trials in this year (HIV prevention and HPV-associated head and neck cancer).

Since the amino acid sequences of these proteins were subjected to computer simulation analysis by online software (NetMHC-4.0 and NetMHCII-2.3) and peptide binding prediction software to localize (map) predicted epitopes of MHC-I and MHC-II, we have identified specific epitopes to be included in candidate vaccines. The B cell epitopes and epitope-rich regions, the sequences of which have homology between SARS-CoV-2 and SARS-CoV-1, were also mapped using on-line software (Bepippred-2.0 and discover).

Four structural proteins (S, N, M and E) of SARS-CoV-2 were each NetMHC 4.0 #https:// services.healthtech.dtu.dk/service.phpNetMHC-4.0) And NetMHCII 2.3%https:// services.healthtech.dtu.dk/service.phpNetMHCII-2.3) Analysis was performed (MHC class I and MHC class II/peptide binding prediction software, respectively).

Using Bepippred 2.0%https://services.healthtech.dtu.dk/service.php？ BepiPred-2.0) Linear B cell epitopes are predicted.

80 HLA class I molecules (for 9-mer peptides) and 54 HLA class II molecules (for 15-mer peptides) were used for T cell epitope prediction.

The described SARS-CoV-1T cell and B cell epitopes are listed and percent homology to SARS-CoV-2 and sequence conservation between different beta-coronaviruses are studied.

The region containing the predicted cluster of B, CD8 and CD4 SARS-CoV-2 epitopes and B, CD and CD4 SARS-CoV-1 epitopes described (and in particular the conserved regions between the different beta-coronaviruses) was selected.

The NAb-targeted SARS-CoV-1 sequence is specifically selected for inclusion in the vaccine region.

The definition of the region takes into account the recently published B cell response profile of SARS-CoV-2 by Dahlke et al.

The regions were also modulated according to predicted epitopes of T cells and B cells published by other groups (Prachar M et al, bioRxiv preprint, bojin F et al preprint, fast et al [7-10 ]).

Based on the above method, we identified the following regions of interest (see table 1).

Table 1: regions of interest identified in SARS-CoV-2 protein that can be used in vaccine design

Example 2

The method comprises the following steps:

expression vectors and methods for protein purification production and quality assurance (including CD40 binding specificity) are described [1;2;3]. UsingCHO transfection kit (Mirus) for protein expression by transient transfection of CHO-S (chinese hamster ovary cells). Cloning used a synthetic DNA cassette encoding (using CHO optimized codons) various SARS-Co-V2 antigen regions, which are typically flanked by restriction sites to facilitate ligation to the vector in seed combination.

[1]Flamar AL.，S.Zurawski，F.Scholz，I.Gayet，L.Ni，XH.Li，E.Klechevsky，J.Quinn，S.Oh，D.H.Kaplan，J.Banchereau and G.Zurawski.2012.Noncovalent assembly of anti-Dendritic Cell antibodies and antigens for evoking immune responses in vitro and in vivo.J.Immunl.189：2645-55.

[2]Flamar AL.，Y.Xue，S.M.Zurawski，M.Montes，B.King，L.Sloan，S.Oh，J.Banchereau，Y.Levy and G.Zurawski.2013.Targeting concatenated HIV antigens to human CD40 expands a broad repertoire of multifunctional CD4+and CD8+T Cells.AIDS.27：2041-51.

[3]Zurawski G.，X.Shen，S.Zurawaski，G.D.Tomaras，D.C.Montafiori，M.Roederer，G.Ferrari，C.Lacabaratz，P.Klucar，Z.Wang，K.E.Foulds，SF.Kao，X.Yu，A.Sato，N.L.Yates，C.LaBranche，S.Stanfield-Oakley，K.Kibler，B.Jacobs，A.Salazar，S.Self，E.Fulp，R.Gottardo，L.Galmin，D.Weiss，A.Cristillo，G.Pantaleo and Y.Levy.2017.Superiority in rhesus macaques of targeting HIV-1 Env gp 140 to CD40 versus LOX-1 in combination with replication-competent NYVAC-KC for induction of Env-specific antibody and T Cell responses.J.Virol.91：1-20.

Results:

we previously obtained the following patents: for a group of human DC receptors: compositions, expression, secretion and methods of use of novel antigen recombinant antibodies to CD40, DCIR, langerhans, and LOX-1. The technology of targeting DCs is based on engineering pathogen antigens directly fused to the C-terminus of the heavy and/or light chain of an anti-DC receptor monoclonal antibody. Thus, the coding sequence of the selected CoV antigen region is fused by molecular cloning to a string of antigen regions (strings-of-occupied) in either the heavy or light chain, separated by an internally defined linker peptide.

Expression constructs of engineered DC-targeting candidate vaccines against covd-19 with various regions or regions encoding SARS-CoV-2 antigen (with or without flexible linker regions) on the C-terminus of the heavy and/or light chain of the humanized anti-human CD40 e12 IgG4 antibody.

The heavy and light chain expression constructs were transiently co-transfected into Expi CHO-S cells and then the expression of the secreted recombinant fusion antibody protein was tested by ELISA to measure the relative yield of the hIgG4 level and/or human CD40 binding. ELISA measurements were fractionated against a 1. Mu.g/ml standard curve of humanized anti-human CD4012E12 IgG4 antibody (see FIG. 4). The initial assay is to determine which combinations of constructs produce products in a range that may be useful for expanding vaccine production. This reflects its productivity as described in table 2.

The quality of the generated candidate vaccine described in table 2 was assessed by: (i) SDS-PAGE analysis: purified rabs. Antigen fusion proteins were resolved by SDS-PAGE under reducing and non-reducing conditions and stained with coomassie blue, and/or (ii) size exclusion chromatography.

Table 2: production ability of constructs

* ELISA CD40 binding secreted products refer to the results of a test in which H chain and L chain expression constructs were co-transfected into the Expi CHO-S cells. Data are broadly classified as none, low, medium or high based on 1 μg/ml of an internal assay control protein (e.g., anti-CD 4012E12 mAb (e.g., anti-human CD4012E12 higgg 4 antibody lot PAB 2220) and/or construct with previously defined expression characteristics, see fig. 4).

* SDS PAGE observations refer to the quality of candidate vaccines for evaluation of production by reduced and/or non-reduced SDS PAGE assessment after purification from CHO-S supernatant by protein a affinity chromatography. Non-finger (usually because expression is none or low, or if identified as e.g. moderate, refers to failure in duplicate or expanded transfection experiments); good qa+ refers to products with H-and L-chain bands, occurring at migration sites consistent with antigen fusion loading, and well-balanced staining consistent with 2h+2l chain structure, with minimal degradation shown by additional lower molecular weight bands; the disadvantages refer to a number of problems (whether alone or in combination) -these include: weak multiple bands QA-or poor fragmentation refers to products that are inconsistent with 2h+2l chain structure and/or have degradation represented by additional lower molecular weight bands; homodimers refer to other lower molecular weight bands that are normally present on non-reduction, consistent with certain 2H products as well as 2h+2l products.

In particular, we generated 8 constructs with optimal productivity and quality, as shown below [ ]FIG. 1)：

anti-CD 40.COVID-19 construct "Gen2a" with 1 peptide (virus SARS-CoV-2 spike-RBD) on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) and 3 peptides (virus SARS-CoV-2Npep2, spep1, spep 4) on the light chain (H anti-CD 40VK 2-LV-hIgGK)

anti-CD 40.COVID-19 construct "Gen2b" with 2 peptides on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) (viruses SARS-CoV-2Spep1, spep 4) and 2 peptides on the light chain (H anti-CD 40VK 2-LV-hIgGK) (viruses SARS-CoV-2Npep2, spike-RBD)

anti-CD 40.COVID-19 construct "Gen2c" with 3 peptides on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) (viruses SARS-CoV-2Npep2, spep1, spep 4) and 1 peptide on the light chain (H anti-CD 40VK 2-LV-hIgGK) (viruses SARS-CoV-2 spike-RBD)

anti-CD 40.COVID-19 construct "CD40.CoV2v" with 1 peptide on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) (virus SARS-CoV-2 spike-RBD) _SA _VAR ) Having 3 peptides (viruses SARS-CoV-2Npep2, spep 1) on the light chain (h anti-CD 40VK 2-LV-hIgGK) _C136S 、Spep4)

anti-CD 40.COVID-19 construct "CD40.N2.RBDv" with 1 peptide (viral SARS-CoV-2Npep 2) on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) and 1 peptide (viral SARS-CoV-2 spike-RBD) on the light chain (H anti-CD 40VK 2-LV-hIgGK) _SA _VAR )

anti-CD 40.COVID-19 construct "CD40.N2.RBDv-2" with 1 peptide (virus SARS-CoV-2 spike-RBD) on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) _SA _VAR ) Having 1 peptide (virus SARS-CoV-2Npep 2) on the light chain (h anti-CD 40VK 2-LV-hIgGK)

anti-CD 40.COVID-19 construct "CD40.RBDvS4.N2" with 2 peptides on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) (virus SARS-CoV-2 spike-RBD) _SA _VAR Spep 4) has 1 peptide (virus SARS-CoV-2Npep 2) on the light chain (h anti-CD 40VK 2-LV-hIgGK)

anti-CD 40.COVID-19 construct "CD40.N2S1.RBDvS4" with 2 peptides on the heavy chain (H anti-CD 40VH3-LV-hIgG 4H) (virus SARS-CoV-2Npep2, spep1 _C136S ) Having 2 peptides (viral SARS-CoV-2 spike-RBD) on the light chain (h anti-CD 40VK 2-LV-hIgGK) _SA _VAR ，Spep4)

We confirmed that anti-CD 40 Gen2a, gen2b and Gen2c fusion proteins bind to human CD40 (FIG. 2). The CD 40-targeted SARS-CoV2pep antigen in Gen2a, gen2b and Gen2c fusion proteins was then tested by in vitro expansion of SARS-CoV-2 specific T cells in a convalescence donor PBMC culture of SARS-CoV-2 infection (FIG. 3).

We show in FIG. 5 that the anti-CD40.Gen2a vaccine elicits a multifunctional CD4+ T cell response in PBMC of patients with convalescence in COVID-19.

Immunogenicity of anti-CD 40 Gen2a vaccines administered in either homologous or heterologous prime/boost vaccination strategies was studied according to the protocol depicted in fig. 6A. The results are shown in fig. 6B. We show that this vaccine elicits S-specific IgG+hu-B cells (FIG. 6B). Interestingly, we showed that S-specific igg+hu-B cells were primed without the addition of an adjuvant (fig. 6B).

Reference is made to:

in the present application, various references describe the state of the art to which the application pertains. The disclosures of these references are incorporated by reference into the present disclosure.

1.Cheng L，Wang Q，Li G，Banga R，Ma J，Yu H，et al.TLR3 agonist and CD40-targeting vaccination induces immune responses and reduces HIV-1 reservoirs.J Clin Invest.2018；128(10)：4387-96.doi：10.1172/JCI99005.

2.Flamar AL，Bonnabau H，Zurawski S，Lacabaratz C，Montes M，Richert L，et al.HIV-1 T cell epitopes targeted to Rhesus macaque CD40 and DCIR：A comparative study of prototype dendritic cell targeting therapeutic vaccine candidates.PLoS One.2018；13(11)：e0207794.doi：10.1371/journal.pone.0207794.

3.Flamar AL，Xue Y，Zurawski SM，Montes M，King B，Sloan L，et al.Targeting concatenated HIV antigens to human CD40 expands a broad repertoire of multifunctional CD4+and CD8+T cells.AIDS.2013；27(13)：2041-51.doi：10.1097/QAD.0b013e3283624305.

4.Flamar AL，Zurawski S，Scholz F，Gayet I，Ni L，Li XH，et al.Noncovalent assembly of anti-dendritic cell antibodies and antigens for evoking immune responses in vitro and in vivo.J Immunol.2012；189(5)：2645-55.dot：10.4049/jimmunol.1102390.

5.Yin W，Duluc D，Joo H，Xue Y，Gu C，Wang Z，et al.Therapeutic HPV Cancer Vaccine Targeted to CD40 Elicits Effective CD8+ T-cell Immunity.Cancer Immunol Res.2016；4(10)：823-34.dot：10.1158/2326-6066.CIR-16-0128.

6.Zurawski G，Shen X，Zurawski S，Tomaras GD，Montefiori DC，Roederer M，et al.Superiority in Rhesus Macaques of Targeting HIV-1 Env gp140 to CD40 versus LOX-1 in Combination with Replication-Competent NYVAC-KC for Induction of Env-Specific Antibody and T Cell Responses.J Virol.2017；91(9).doi：10.1128/JVI.01596-16.

7.Baruah V，Bose S.Immunoinformatics-aided identification of T cell and B cell epitopes in the surface glycoprotein of 2019-nCoV.J Med Virol.2020；92(5)：495-500.doi：10.1002/jmv.25698.

8.Lee CH，Koohy H.In silico identification of vaccine targets for 2019-nCoV.F1000Res.2020；9：145.doi：10.12688/f1000research.22507.2.

9.Ahmed SF，Quadeer AA，McKay MR.Preliminary Identification of Potential Vaccine Targets for the COVID-19 Coronavirus(SARS-CoV-2)Based on SARS-CoV Immunological Studies.Viruses.2020；12(3).doi：10.3390/v12030254.

10.Grifoni A，Sidney J，Zhang Y，Scheuermann RH，Peters B，Sette A.A Sequence Homology and Bioinformatic Approach Can Predict Candidate Targets for Immune Responses to SARS-CoV-2.Cell Host Microbe.2020；27(4)：671-80 e2.doi：10.1016/j.chom.2020.03.002.

Sequence listing

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Leu Glu Gln Trp Asn Leu Val Ile Gly Phe Leu Phe Leu Thr Trp Ile

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Cys Leu Leu Gln Phe Ala Tyr Ala Asn Arg Asn Arg Phe Leu Tyr Ile

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Ile Lys Leu Ile Phe Leu Trp Leu Leu Trp Pro Val Thr Leu Ala Cys

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Phe Val Leu Ala Ala Val Tyr Arg Ile Asn Trp Ile Thr Gly Gly Ile

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Ala Ile Ala Met Ala Cys Leu Val Gly Leu Met Trp Leu Ser Tyr Phe

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Ile Ala Ser Phe Arg Leu Phe Ala Arg Thr Arg Ser Met Trp Ser Phe

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Asn Pro Glu Thr Asn Ile Leu Leu Asn Val Pro Leu His Gly Thr Ile

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Leu Thr Arg Pro Leu Leu Glu Ser Glu Leu Val Ile Gly Ala Val Ile

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Leu Arg Gly His Leu Arg Ile Ala Gly His His Leu Gly Arg Cys Asp

145 150 155 160

Ile Lys Asp Leu Pro Lys Glu Ile Thr Val Ala Thr Ser Arg Thr Leu

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Ser Tyr Tyr Lys Leu Gly Ala Ser Gln Arg Val Ala Gly Asp Ser Gly

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Phe Ala Ala Tyr Ser Arg Tyr Arg Ile Gly Asn Tyr Lys Leu Asn Thr

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Asp His Ser Ser Ser Ser Asp Asn Ile Ala Leu Leu Val Gln

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Met Ser Asp Asn Gly Pro Gln Asn Gln Arg Asn Ala Pro Arg Ile Thr

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Phe Gly Gly Pro Ser Asp Ser Thr Gly Ser Asn Gln Asn Gly Glu Arg

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Ser Gly Ala Arg Ser Lys Gln Arg Arg Pro Gln Gly Leu Pro Asn Asn

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Thr Ala Ser Trp Phe Thr Ala Leu Thr Gln His Gly Lys Glu Asp Leu

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Lys Phe Pro Arg Gly Gln Gly Val Pro Ile Asn Thr Asn Ser Ser Pro

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Asp Asp Gln Ile Gly Tyr Tyr Arg Arg Ala Thr Arg Arg Ile Arg Gly

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Gly Asp Gly Lys Met Lys Asp Leu Ser Pro Arg Trp Tyr Phe Tyr Tyr

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Leu Gly Thr Gly Pro Glu Ala Gly Leu Pro Tyr Gly Ala Asn Lys Asp

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Gly Ile Ile Trp Val Ala Thr Glu Gly Ala Leu Asn Thr Pro Lys Asp

130 135 140

His Ile Gly Thr Arg Asn Pro Ala Asn Asn Ala Ala Ile Val Leu Gln

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Leu Pro Gln Gly Thr Thr Leu Pro Lys Gly Phe Tyr Ala Glu Gly Ser

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Arg Gly Gly Ser Gln Ala Ser Ser Arg Ser Ser Ser Arg Ser Arg Asn

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Ser Ser Arg Asn Ser Thr Pro Gly Ser Ser Arg Gly Thr Ser Pro Ala

195 200 205

Arg Met Ala Gly Asn Gly Gly Asp Ala Ala Leu Ala Leu Leu Leu Leu

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Asp Arg Leu Asn Gln Leu Glu Ser Lys Met Ser Gly Lys Gly Gln Gln

225 230 235 240

Gln Gln Gly Gln Thr Val Thr Lys Lys Ser Ala Ala Glu Ala Ser Lys

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Lys Pro Arg Gln Lys Arg Thr Ala Thr Lys Ala Tyr Asn Val Thr Gln

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Ala Phe Gly Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp

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Gln Glu Leu Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile

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Ala Gln Phe Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile

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Gly Met Glu Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala

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Ile Lys Leu Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu

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Leu Asn Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro

355 360 365

Lys Lys Asp Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln

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Arg Gln Lys Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu

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Asp Asp Phe Ser Lys Gln Leu Gln Gln Ser Met Ser Ser Ala Asp Ser

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Thr Gln Ala

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Met Phe Val Phe Leu Val Leu Leu Pro Leu Val Ser Ser Gln Cys Val

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Asn Leu Thr Thr Arg Thr Gln Leu Pro Pro Ala Tyr Thr Asn Ser Phe

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Thr Arg Gly Val Tyr Tyr Pro Asp Lys Val Phe Arg Ser Ser Val Leu

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His Ser Thr Gln Asp Leu Phe Leu Pro Phe Phe Ser Asn Val Thr Trp

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Phe His Ala Ile His Val Ser Gly Thr Asn Gly Thr Lys Arg Phe Asp

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Asn Pro Val Leu Pro Phe Asn Asp Gly Val Tyr Phe Ala Ser Thr Glu

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Lys Ser Asn Ile Ile Arg Gly Trp Ile Phe Gly Thr Thr Leu Asp Ser

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Lys Thr Gln Ser Leu Leu Ile Val Asn Asn Ala Thr Asn Val Val Ile

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Lys Val Cys Glu Phe Gln Phe Cys Asn Asp Pro Phe Leu Gly Val Tyr

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Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr

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Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu

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Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe

180 185 190

Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr

195 200 205

Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu

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Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr

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Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Pro Gly Asp Ser Ser Ser

245 250 255

Gly Trp Thr Ala Gly Ala Ala Ala Tyr Tyr Val Gly Tyr Leu Gln Pro

260 265 270

Arg Thr Phe Leu Leu Lys Tyr Asn Glu Asn Gly Thr Ile Thr Asp Ala

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Val Asp Cys Ala Leu Asp Pro Leu Ser Glu Thr Lys Cys Thr Leu Lys

290 295 300

Ser Phe Thr Val Glu Lys Gly Ile Tyr Gln Thr Ser Asn Phe Arg Val

305 310 315 320

Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys

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Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala

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Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu

355 360 365

Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro

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Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe

385 390 395 400

Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly

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Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys

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Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn

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Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe

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Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys

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Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly

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Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val

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Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys

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Lys Ser Thr Asn Leu Val Lys Asn Lys Cys Val Asn Phe Asn Phe Asn

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Gly Leu Thr Gly Thr Gly Val Leu Thr Glu Ser Asn Lys Lys Phe Leu

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Pro Phe Gln Gln Phe Gly Arg Asp Ile Ala Asp Thr Thr Asp Ala Val

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Arg Asp Pro Gln Thr Leu Glu Ile Leu Asp Ile Thr Pro Cys Ser Phe

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Gly Gly Val Ser Val Ile Thr Pro Gly Thr Asn Thr Ser Asn Gln Val

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Ala Val Leu Tyr Gln Asp Val Asn Cys Thr Glu Val Pro Val Ala Ile

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His Ala Asp Gln Leu Thr Pro Thr Trp Arg Val Tyr Ser Thr Gly Ser

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Asn Val Phe Gln Thr Arg Ala Gly Cys Leu Ile Gly Ala Glu His Val

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Asn Asn Ser Tyr Glu Cys Asp Ile Pro Ile Gly Ala Gly Ile Cys Ala

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Ser Tyr Gln Thr Gln Thr Asn Ser Pro Arg Arg Ala Arg Ser Val Ala

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Ser Gln Ser Ile Ile Ala Tyr Thr Met Ser Leu Gly Ala Glu Asn Ser

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Val Ala Tyr Ser Asn Asn Ser Ile Ala Ile Pro Thr Asn Phe Thr Ile

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Ser Val Thr Thr Glu Ile Leu Pro Val Ser Met Thr Lys Thr Ser Val

725 730 735

Asp Cys Thr Met Tyr Ile Cys Gly Asp Ser Thr Glu Cys Ser Asn Leu

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Leu Leu Gln Tyr Gly Ser Phe Cys Thr Gln Leu Asn Arg Ala Leu Thr

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Gly Ile Ala Val Glu Gln Asp Lys Asn Thr Gln Glu Val Phe Ala Gln

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Val Lys Gln Ile Tyr Lys Thr Pro Pro Ile Lys Asp Phe Gly Gly Phe

785 790 795 800

Asn Phe Ser Gln Ile Leu Pro Asp Pro Ser Lys Pro Ser Lys Arg Ser

805 810 815

Phe Ile Glu Asp Leu Leu Phe Asn Lys Val Thr Leu Ala Asp Ala Gly

820 825 830

Phe Ile Lys Gln Tyr Gly Asp Cys Leu Gly Asp Ile Ala Ala Arg Asp

835 840 845

Leu Ile Cys Ala Gln Lys Phe Asn Gly Leu Thr Val Leu Pro Pro Leu

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Leu Thr Asp Glu Met Ile Ala Gln Tyr Thr Ser Ala Leu Leu Ala Gly

865 870 875 880

Thr Ile Thr Ser Gly Trp Thr Phe Gly Ala Gly Ala Ala Leu Gln Ile

885 890 895

Pro Phe Ala Met Gln Met Ala Tyr Arg Phe Asn Gly Ile Gly Val Thr

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Gln Asn Val Leu Tyr Glu Asn Gln Lys Leu Ile Ala Asn Gln Phe Asn

915 920 925

Ser Ala Ile Gly Lys Ile Gln Asp Ser Leu Ser Ser Thr Ala Ser Ala

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Leu Gly Lys Leu Gln Asp Val Val Asn Gln Asn Ala Gln Ala Leu Asn

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Thr Leu Val Lys Gln Leu Ser Ser Asn Phe Gly Ala Ile Ser Ser Val

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Leu Asn Asp Ile Leu Ser Arg Leu Asp Lys Val Glu Ala Glu Val Gln

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Ile Asp Arg Leu Ile Thr Gly Arg Leu Gln Ser Leu Gln Thr Tyr Val

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Thr Gln Gln Leu Ile Arg Ala Ala Glu Ile Arg Ala Ser Ala Asn

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Leu Ala Ala Thr Lys Met Ser Glu Cys Val Leu Gly Gln Ser Lys

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Arg Val Asp Phe Cys Gly Lys Gly Tyr His Leu Met Ser Phe Pro

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Gln Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val

1055 1060 1065

Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His

1070 1075 1080

Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

1085 1090 1095

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln

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Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val

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Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

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Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

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His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn

1160 1165 1170

Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu

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Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu

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Gly Lys Tyr Glu Gln Tyr Ile Lys Trp Pro Trp Tyr Ile Trp Leu

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Gly Phe Ile Ala Gly Leu Ile Ala Ile Val Met Val Thr Ile Met

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Leu Cys Cys Met Thr Ser Cys Cys Ser Cys Leu Lys Gly Cys Cys

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Ser Cys Gly Ser Cys Cys Lys Phe Asp Glu Asp Asp Ser Glu Pro

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Val Leu Lys Gly Val Lys Leu His Tyr Thr

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Met Gln Lys Gly Asp Gln Asn Pro Gln Ile Ala Ala His Val Ile Ser

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Glu Ala Ser Ser Lys Thr Thr Ser Val Leu Gln Trp Ala Glu Lys Gly

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Tyr Tyr Thr Met Ser Asn Asn Leu Val Thr Leu Glu Asn Gly Lys Gln

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Leu Thr Val Lys Arg Gln Gly Leu Tyr Tyr Ile Tyr Ala Gln Val Thr

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Phe Cys Ser Asn Arg Glu Ala Ser Ser Gln Ala Pro Phe Ile Ala Ser

65 70 75 80

Leu Cys Leu Lys Ser Pro Gly Arg Phe Glu Arg Ile Leu Leu Arg Ala

85 90 95

Ala Asn Thr His Ser Ser Ala Lys Pro Cys Gly Gln Gln Ser Ile His

100 105 110

Leu Gly Gly Val Phe Glu Leu Gln Pro Gly Ala Ser Val Phe Val Asn

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Val Thr Asp Pro Ser Gln Val Ser His Gly Thr Gly Phe Thr Ser Phe

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Gly Leu Leu Lys Leu

145

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<223> Npep2-Spep1-f2-Spep4 Multi-epitope Polypeptides

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Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp Gln Glu Leu

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Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile Ala Gln Phe

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Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile Gly Met Glu

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Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala Ile Lys Leu

50 55 60

Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu Leu Asn Lys

65 70 75 80

His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro Lys Lys Asp

85 90 95

Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln Arg Gln Lys

100 105 110

Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu Asp Asp Phe

115 120 125

Ser Lys Gln Leu Gln Gln Ser Met Ala Arg Asn Val Val Ile Lys Val

130 135 140

Cys Glu Phe Gln Phe Ser Asn Asp Pro Phe Leu Gly Val Tyr Tyr His

145 150 155 160

Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser

165 170 175

Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp

180 185 190

Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe

195 200 205

Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile

210 215 220

Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu

225 230 235 240

Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu

245 250 255

Ala Leu His Arg Ser Tyr Leu Thr Arg Pro Thr Ser Thr Pro Ala Asp

260 265 270

Ser Ser Thr Ile Thr Pro Thr Ala Thr Pro Thr Ala Thr Pro Thr Ile

275 280 285

Lys Gly Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro

290 295 300

Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly

305 310 315 320

Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His

325 330 335

Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr

340 345 350

Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val

355 360 365

Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys

370 375 380

Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp

385 390 395 400

Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys

405 410 415

Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu

420 425 430

Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr

435 440

<210> 6

<211> 306

<212> PRT

<213> artificial sequence

<220>

<223> Spep1-f2-Spep4

<400> 6

Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe Ser Asn Asp Pro Phe

1 5 10 15

Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu

20 25 30

Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser

35 40 45

Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn

50 55 60

Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr

65 70 75 80

Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe

85 90 95

Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr

100 105 110

Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Arg Pro

115 120 125

Thr Ser Thr Pro Ala Asp Ser Ser Thr Ile Thr Pro Thr Ala Thr Pro

130 135 140

Thr Ala Thr Pro Thr Ile Lys Gly Ala Pro His Gly Val Val Phe Leu

145 150 155 160

His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro

165 170 175

Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe

180 185 190

Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu

195 200 205

Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp

210 215 220

Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro

225 230 235 240

Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His

245 250 255

Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser

260 265 270

Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys

275 280 285

Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu

290 295 300

Gln Tyr

305

<210> 7

<211> 361

<212> PRT

<213> artificial sequence

<220>

<223> Npep2-RBD

<400> 7

Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp Gln Glu Leu

1 5 10 15

Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile Ala Gln Phe

20 25 30

Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile Gly Met Glu

35 40 45

Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala Ile Lys Leu

50 55 60

Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu Leu Asn Lys

65 70 75 80

His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro Lys Lys Asp

85 90 95

Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln Arg Gln Lys

100 105 110

Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu Asp Asp Phe

115 120 125

Ser Lys Gln Leu Gln Gln Ser Met Ala Ser Arg Val Gln Pro Thr Glu

130 135 140

Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu

145 150 155 160

Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys

165 170 175

Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala

180 185 190

Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn

195 200 205

Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly

210 215 220

Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp

225 230 235 240

Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp

245 250 255

Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu

260 265 270

Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile

275 280 285

Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu

290 295 300

Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr

305 310 315 320

Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu

325 330 335

Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn

340 345 350

Leu Val Lys Asn Lys Ser Val Asn Phe

355 360

<210> 8

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR1

<400> 8

Gly Phe Thr Phe Ser Asp Tyr Tyr Met Tyr

1 5 10

<210> 9

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR2

<400> 9

Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val Lys

1 5 10 15

Gly

<210> 10

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR3

<400> 10

Arg Gly Leu Pro Phe His Ala Met Asp Tyr

1 5 10

<210> 11

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR1

<400> 11

Ser Ala Ser Gln Gly Ile Ser Asn Tyr Leu Asn

1 5 10

<210> 12

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR2

<400> 12

Tyr Thr Ser Ile Leu His Ser

1 5

<210> 13

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR3

<400> 13

Gln Gln Phe Asn Lys Leu Pro Pro Thr

1 5

<210> 14

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR1

<400> 14

Gly Tyr Ser Phe Thr Gly Tyr Tyr Met His

1 5 10

<210> 15

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR2

<400> 15

Arg Ile Asn Pro Tyr Asn Gly Ala Thr Ser Tyr Asn Gln Asn Phe Lys

1 5 10 15

Asp

<210> 16

<211> 5

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR3

<400> 16

Glu Asp Tyr Val Tyr

1 5

<210> 17

<211> 16

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR1

<400> 17

Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly Asn Thr Tyr Leu His

1 5 10 15

<210> 18

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR2

<400> 18

Lys Val Ser Asn Arg Phe Ser

1 5

<210> 19

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR3

<400> 19

Ser Gln Ser Thr His Val Pro Trp Thr

1 5

<210> 20

<211> 10

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR1

<400> 20

Gly Tyr Thr Phe Thr Asp Tyr Val Leu His

1 5 10

<210> 21

<211> 17

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR2

<400> 21

Tyr Ile Asn Pro Tyr Asn Asp Gly Thr Lys Tyr Asn Glu Lys Phe Lys

1 5 10 15

Gly

<210> 22

<211> 12

<212> PRT

<213> artificial sequence

<220>

<223> H-CDR3

<400> 22

Gly Tyr Pro Ala Tyr Ser Gly Tyr Ala Met Asp Tyr

1 5 10

<210> 23

<211> 11

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR1

<400> 23

Arg Ala Ser Gln Asp Ile Ser Asn Tyr Leu Asn

1 5 10

<210> 24

<211> 7

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR2

<400> 24

Tyr Thr Ser Arg Leu His Ser

1 5

<210> 25

<211> 9

<212> PRT

<213> artificial sequence

<220>

<223> L-CDR3

<400> 25

His His Gly Asn Thr Leu Pro Trp Thr

1 5

<210> 26

<211> 116

<212> PRT

<213> artificial sequence

<220>

<223> VH Domain

<400> 26

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr

20 25 30

Tyr Met His Trp Val Lys Gln Ala His Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Arg Ile Asn Pro Tyr Asn Gly Ala Thr Ser Tyr Asn Gln Asn Phe

50 55 60

Lys Asp Arg Ala Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Asp Tyr Val Tyr Trp Gly Gln Gly Thr Thr Val Thr Val

100 105 110

Ser Ser Ala Ser

115

<210> 27

<211> 108

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 27

Asp Val Val Met Thr Gln Ser Pro Leu Ser Leu Pro Val Thr Leu Gly

1 5 10 15

Gln Pro Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser

20 25 30

Asn Gly Asn Thr Tyr Leu His Trp Tyr Gln Gln Arg Pro Gly Gln Ser

35 40 45

Pro Arg Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro

50 55 60

Asp Arg Phe Ser Gly Ser Gly Ser Gly Thr Asp Phe Thr Leu Lys Ile

65 70 75 80

Ser Arg Val Glu Ala Glu Asp Val Gly Val Tyr Phe Cys Ser Gln Ser

85 90 95

Thr His Val Pro Trp Thr Phe Gly Gly Gly Thr Lys

100 105

<210> 28

<211> 116

<212> PRT

<213> artificial sequence

<220>

<223> VH Domain

<400> 28

Glu Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Ser Phe Thr Gly Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Arg Ile Asn Pro Tyr Asn Gly Ala Thr Ser Tyr Asn Gln Asn Phe

50 55 60

Lys Asp Arg Val Thr Leu Thr Val Asp Lys Ser Thr Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Arg Ser Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Glu Asp Tyr Val Tyr Trp Gly Gln Gly Thr Thr Val Thr Val

100 105 110

Ser Ser Ala Ser

115

<210> 29

<211> 123

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 29

Glu Val Gln Leu Gln Gln Ser Gly Pro Glu Leu Val Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

20 25 30

Val Leu His Trp Val Lys Gln Lys Pro Gly Gln Gly Leu Glu Trp Ile

35 40 45

Gly Tyr Ile Asn Pro Tyr Asn Asp Gly Thr Lys Tyr Asn Glu Lys Phe

50 55 60

Lys Gly Lys Ala Thr Leu Thr Ser Asp Lys Ser Ser Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Gly Tyr Pro Ala Tyr Ser Gly Tyr Ala Met Asp Tyr Trp Gly

100 105 110

Gln Gly Thr Ser Val Thr Val Ser Ser Ala Ser

115 120

<210> 30

<211> 103

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 30

Asp Ile Gln Met Thr Gln Thr Thr Ser Ser Leu Ser Ala Ser Leu Gly

1 5 10 15

Asp Arg Val Thr Ile Ser Cys Arg Ala Ser Gln Asp Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Asp Gly Thr Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Arg Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Ser Leu Thr Ile Ser Asn Leu Glu Gln

65 70 75 80

Glu Asp Ile Ala Thr Tyr Phe Cys His His Gly Asn Thr Leu Pro Trp

85 90 95

Thr Phe Gly Gly Gly Thr Lys

100

<210> 31

<211> 121

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 31

Asp Val Gln Leu Gln Glu Ser Gly Pro Asp Leu Val Lys Pro Ser Gln

1 5 10 15

Ser Leu Ser Leu Thr Cys Thr Val Thr Gly Tyr Ser Ile Thr Ser Asp

20 25 30

Tyr Ser Trp His Trp Ile Arg Gln Phe Pro Gly Asn Lys Leu Glu Trp

35 40 45

Met Gly Tyr Ile Tyr Tyr Ser Gly Ser Thr Asn Tyr Asn Pro Ser Leu

50 55 60

Lys Ser Arg Ile Ser Ile Thr Arg Asp Thr Ser Lys Asn Gln Phe Phe

65 70 75 80

Leu Gln Leu Asn Ser Val Thr Thr Glu Asp Ser Ala Thr Tyr Phe Cys

85 90 95

Ala Arg Phe Tyr Tyr Gly Tyr Ser Phe Phe Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Thr Leu Thr Val Ser Ser Ala Ser

115 120

<210> 32

<211> 102

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 32

Gln Ile Val Leu Thr Gln Ser Pro Ala Phe Met Ser Ala Ser Pro Gly

1 5 10 15

Glu Lys Val Thr Met Thr Cys Ser Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

His Trp Tyr Gln Gln Lys Ser Gly Thr Ser Pro Lys Arg Trp Ile Tyr

35 40 45

Asp Thr Ser Lys Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Ser Met Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Ala Gly Thr Lys

100

<210> 33

<211> 128

<212> PRT

<213> artificial sequence

<220>

<223> VH Domain

<400> 33

Gln Val Gln Leu Val Gln Ser Gly Ala Glu Val Lys Lys Pro Gly Ala

1 5 10 15

Ser Val Lys Val Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr

20 25 30

Tyr Met His Trp Val Arg Gln Ala Pro Gly Gln Gly Leu Glu Trp Met

35 40 45

Gly Trp Ile Asn Pro Asp Ser Gly Gly Thr Asn Tyr Ala Gln Lys Phe

50 55 60

Gln Gly Arg Val Thr Met Thr Arg Asp Thr Ser Ile Ser Thr Ala Tyr

65 70 75 80

Met Glu Leu Asn Arg Leu Arg Ser Asp Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Asp Gln Pro Leu Gly Tyr Cys Thr Asn Gly Val Cys Ser Tyr

100 105 110

Phe Asp Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ser Ala Ser

115 120 125

<210> 34

<211> 103

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 34

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Val Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Arg Ala Ser Gln Gly Ile Tyr Ser Trp

20 25 30

Leu Ala Trp Tyr Gln Gln Lys Pro Gly Lys Ala Pro Asn Leu Leu Ile

35 40 45

Tyr Thr Ala Ser Thr Leu Gln Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Phe Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Ala Asn Ile Phe Pro Leu

85 90 95

Thr Phe Gly Gly Gly Thr Lys

100

<210> 35

<211> 121

<212> PRT

<213> artificial sequence

<220>

<223> VH Domain

<400> 35

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser

115 120

<210> 36

<211> 103

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 36

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys

100

<210> 37

<211> 81

<212> PRT

<213> artificial sequence

<220>

<223> VH Domain

<400> 37

Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp Tyr

1 5 10 15

Val Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp Ile

20 25 30

Gly Asp Ile Tyr Pro Gly Ser Gly Tyr Ser Phe Tyr Asn Glu Asn Phe

35 40 45

Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Thr Thr Ala Tyr

50 55 60

Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe Cys

65 70 75 80

Ala

<210> 38

<211> 76

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 38

Ala Ser Ile Ser Cys Arg Ser Ser Gln Ser Leu Val His Ser Asn Gly

1 5 10 15

Asn Thr Tyr Leu His Trp Tyr Leu Gln Lys Pro Gly Gln Ser Pro Lys

20 25 30

Leu Leu Ile Tyr Lys Val Ser Asn Arg Phe Ser Gly Val Pro Asp Arg

35 40 45

Phe Ser Gly Ser Gly Ser Gly Thr Asn Phe Thr Leu Lys Ile Ser Arg

50 55 60

Val Glu Ala Glu Asp Leu Gly Leu Tyr Phe Cys Ser

65 70 75

<210> 39

<211> 82

<212> PRT

<213> artificial sequence

<220>

<223> VH Domain

<400> 39

Ser Ser Val Lys Met Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Asp

1 5 10 15

Tyr Val Ile Ser Trp Val Lys Gln Arg Thr Gly Gln Gly Leu Glu Trp

20 25 30

Ile Gly Asp Ile Tyr Pro Gly Ser Gly Tyr Ser Phe Tyr Asn Glu Asn

35 40 45

Phe Lys Gly Lys Ala Thr Leu Thr Ala Asp Lys Ser Ser Thr Thr Ala

50 55 60

Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Phe

65 70 75 80

Cys Ala

<210> 40

<211> 74

<212> PRT

<213> artificial sequence

<220>

<223> VL Domain

<400> 40

Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser Asn

1 5 10 15

Tyr Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly Leu

20 25 30

Ile Gly Gly Thr Asn Asn Arg Val Ser Gly Val Pro Ala Arg Phe Ser

35 40 45

Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala Gln

50 55 60

Thr Glu Asp Glu Ala Ile Tyr Phe Cys Ala

65 70

<210> 41

<211> 456

<212> PRT

<213> artificial sequence

<220>

<223> heavy chain

<400> 41

Gln Val Gln Leu Gln Gln Ser Gly Ala Glu Leu Val Arg Pro Gly Ala

1 5 10 15

Ser Val Thr Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Ile Asp His

20 25 30

Asp Met His Trp Val Gln Gln Thr Pro Val Tyr Gly Leu Glu Trp Ile

35 40 45

Gly Ala Ile Asp Pro Glu Thr Gly Asp Thr Gly Tyr Asn Gln Lys Phe

50 55 60

Lys Gly Lys Ala Ile Leu Thr Ala Asp Lys Ser Ser Arg Thr Ala Tyr

65 70 75 80

Met Glu Leu Arg Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys

85 90 95

Thr Ile Pro Phe Tyr Tyr Ser Asn Tyr Ser Pro Phe Ala Tyr Trp Gly

100 105 110

Gln Gly Ala Leu Val Thr Val Ser Ala Ala Lys Thr Thr Ala Pro Ser

115 120 125

Val Tyr Pro Leu Ala Pro Val Cys Gly Gly Thr Thr Gly Ser Ser Val

130 135 140

Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Pro Val Thr Leu

145 150 155 160

Thr Trp Asn Ser Gly Ser Leu Ser Ser Gly Val His Thr Phe Pro Ala

165 170 175

Leu Leu Gln Ser Gly Leu Tyr Thr Leu Ser Ser Ser Val Thr Val Thr

180 185 190

Ser Asn Thr Trp Pro Ser Gln Thr Ile Thr Cys Asn Val Ala His Pro

195 200 205

Ala Ser Ser Thr Lys Val Asp Lys Lys Ile Glu Pro Arg Val Pro Ile

210 215 220

Thr Gln Asn Pro Cys Pro Pro Leu Lys Glu Cys Pro Pro Cys Ala Asp

225 230 235 240

Leu Leu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Lys Ile Lys Asp

245 250 255

Val Leu Met Ile Ser Leu Ser Pro Met Val Thr Cys Val Val Val Asp

260 265 270

Val Ser Glu Asp Asp Pro Asp Ala Gln Ile Ser Trp Phe Val Asn Asn

275 280 285

Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu Asp Tyr Asn

290 295 300

Ser Thr Leu Arg Val Val Ser Ala Leu Pro Ile Gln His Gln Asp Trp

305 310 315 320

Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Arg Ala Leu Pro

325 330 335

Ser Pro Ile Glu Lys Thr Ile Ser Lys Pro Arg Gly Pro Val Arg Ala

340 345 350

Pro Gln Val Tyr Val Leu Pro Pro Pro Ala Glu Glu Met Thr Lys Lys

355 360 365

Glu Phe Ser Leu Thr Cys Met Ile Thr Gly Phe Leu Pro Ala Glu Ile

370 375 380

Ala Val Asp Trp Thr Ser Asn Gly Arg Thr Glu Gln Asn Tyr Lys Asn

385 390 395 400

Thr Ala Thr Val Leu Asp Ser Asp Gly Ser Tyr Phe Met Tyr Ser Lys

405 410 415

Leu Arg Val Gln Lys Ser Thr Trp Glu Arg Gly Ser Leu Phe Ala Cys

420 425 430

Ser Val Val His Glu Gly Leu His Asn His Leu Thr Thr Lys Thr Ile

435 440 445

Ser Arg Ser Leu Gly Lys Ala Ser

450 455

<210> 42

<211> 213

<212> PRT

<213> artificial sequence

<220>

<223> light chain

<400> 42

Gln Ile Val Leu Ser Gln Ser Pro Ala Ile Leu Ser Ala Ser Pro Gly

1 5 10 15

Glu Lys Val Thr Met Thr Cys Arg Ala Ser Ser Ser Val Ser Tyr Met

20 25 30

His Trp Tyr Gln Arg Lys Pro Gly Ser Ser Pro Lys Pro Trp Ile Tyr

35 40 45

Ala Thr Ser Asn Leu Ala Ser Gly Val Pro Ala Arg Phe Ser Gly Ser

50 55 60

Gly Ser Gly Thr Ser Tyr Ser Leu Thr Ile Ser Arg Val Glu Ala Glu

65 70 75 80

Asp Ala Ala Thr Tyr Tyr Cys Gln Gln Trp Ser Ser Asn Pro Leu Thr

85 90 95

Phe Gly Ala Gly Thr Lys Leu Glu Leu Lys Arg Ala Asp Ala Ala Pro

100 105 110

Thr Val Ser Ile Phe Pro Pro Ser Ser Glu Gln Leu Thr Ser Gly Gly

115 120 125

Ala Ser Val Val Cys Phe Leu Asn Asn Phe Tyr Pro Lys Asp Ile Asn

130 135 140

Val Lys Trp Lys Ile Asp Gly Ser Glu Arg Gln Asn Gly Val Leu Asn

145 150 155 160

Ser Trp Thr Asp Gln Asp Ser Lys Asp Ser Thr Tyr Ser Met Ser Ser

165 170 175

Thr Leu Thr Leu Thr Lys Asp Glu Tyr Glu Arg His Asn Ser Tyr Thr

180 185 190

Cys Glu Ala Thr His Lys Thr Ser Thr Ser Pro Ile Val Lys Ser Phe

195 200 205

Asn Arg Asn Glu Cys

210

<210> 43

<211> 27

<212> PRT

<213> artificial sequence

<220>

<223> joint

<400> 43

Gln Thr Pro Thr Asn Thr Ile Ser Val Thr Pro Thr Asn Asn Ser Thr

1 5 10 15

Pro Thr Asn Asn Ser Asn Pro Lys Pro Asn Pro

20 25

<210> 44

<211> 25

<212> PRT

<213> artificial sequence

<220>

<223> joint

<400> 44

Ser Ser Val Ser Pro Thr Thr Ser Val His Pro Thr Pro Thr Ser Val

1 5 10 15

Pro Pro Thr Pro Thr Lys Ser Ser Pro

20 25

<210> 45

<211> 25

<212> PRT

<213> artificial sequence

<220>

<223> joint

<400> 45

Pro Thr Ser Thr Pro Ala Asp Ser Ser Thr Ile Thr Pro Thr Ala Thr

1 5 10 15

Pro Thr Ala Thr Pro Thr Ile Lys Gly

20 25

<210> 46

<211> 25

<212> PRT

<213> artificial sequence

<220>

<223> joint

<400> 46

Thr Val Thr Pro Thr Ala Thr Ala Thr Pro Ser Ala Ile Val Thr Thr

1 5 10 15

Ile Thr Pro Thr Ala Thr Thr Lys Pro

20 25

<210> 47

<211> 25

<212> PRT

<213> artificial sequence

<220>

<223> joint

<400> 47

Thr Asn Gly Ser Ile Thr Val Ala Ala Thr Ala Pro Thr Val Thr Pro

1 5 10 15

Thr Val Asn Ala Thr Pro Ser Ala Ala

20 25

<210> 48

<211> 671

<212> PRT

<213> artificial sequence

<220>

<223> heavy chain fused to antigen

<400> 48

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ala Ser

435 440 445

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn

450 455 460

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

465 470 475 480

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

485 490 495

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

500 505 510

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

515 520 525

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

530 535 540

Thr Gly Lys Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

545 550 555 560

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

565 570 575

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

580 585 590

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

595 600 605

Pro Cys Asn Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

610 615 620

Tyr Gly Phe Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val

625 630 635 640

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

645 650 655

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Ser Val Asn Phe

660 665 670

<210> 49

<211> 687

<212> PRT

<213> artificial sequence

<220>

<223> light chain fused to antigen

<400> 49

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys Ala Ser Thr Asn Gly Ser Ile Thr Val Ala

210 215 220

Ala Thr Ala Pro Thr Val Thr Pro Thr Val Asn Ala Thr Pro Ser Ala

225 230 235 240

Ala Ala Ser Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp

245 250 255

Gln Glu Leu Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile

260 265 270

Ala Gln Phe Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile

275 280 285

Gly Met Glu Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala

290 295 300

Ile Lys Leu Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu

305 310 315 320

Leu Asn Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro

325 330 335

Lys Lys Asp Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln

340 345 350

Arg Gln Lys Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu

355 360 365

Asp Asp Phe Ser Lys Gln Leu Gln Gln Ser Met Ala Arg Asn Val Val

370 375 380

Ile Lys Val Cys Glu Phe Gln Phe Ser Asn Asp Pro Phe Leu Gly Val

385 390 395 400

Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val

405 410 415

Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe

420 425 430

Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu

435 440 445

Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His

450 455 460

Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu

465 470 475 480

Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln

485 490 495

Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Arg Pro Thr Ser Thr

500 505 510

Pro Ala Asp Ser Ser Thr Ile Thr Pro Thr Ala Thr Pro Thr Ala Thr

515 520 525

Pro Thr Ile Lys Gly Ala Pro His Gly Val Val Phe Leu His Val Thr

530 535 540

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys

545 550 555 560

His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

565 570 575

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile

580 585 590

Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile

595 600 605

Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp

610 615 620

Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro

625 630 635 640

Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn

645 650 655

Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn

660 665 670

Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr

675 680 685

<210> 50

<211> 783

<212> PRT

<213> artificial sequence

<220>

<223> heavy chain fused to antigen

<400> 50

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ala Ser

435 440 445

Gln Thr Pro Thr Asn Thr Ile Ser Val Thr Pro Thr Asn Asn Ser Thr

450 455 460

Pro Thr Asn Asn Ser Asn Pro Lys Pro Asn Pro Ala Arg Asn Val Val

465 470 475 480

Ile Lys Val Cys Glu Phe Gln Phe Ser Asn Asp Pro Phe Leu Gly Val

485 490 495

Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val

500 505 510

Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe

515 520 525

Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu

530 535 540

Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His

545 550 555 560

Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu

565 570 575

Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln

580 585 590

Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Arg Pro Thr Ser Thr

595 600 605

Pro Ala Asp Ser Ser Thr Ile Thr Pro Thr Ala Thr Pro Thr Ala Thr

610 615 620

Pro Thr Ile Lys Gly Ala Pro His Gly Val Val Phe Leu His Val Thr

625 630 635 640

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys

645 650 655

His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

660 665 670

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile

675 680 685

Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile

690 695 700

Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp

705 710 715 720

Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro

725 730 735

Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn

740 745 750

Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn

755 760 765

Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr

770 775 780

<210> 51

<211> 604

<212> PRT

<213> artificial sequence

<220>

<223> light chain fused to antigen

<400> 51

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys Ala Ser Thr Asn Gly Ser Ile Thr Val Ala

210 215 220

Ala Thr Ala Pro Thr Val Thr Pro Thr Val Asn Ala Thr Pro Ser Ala

225 230 235 240

Ala Ala Ser Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp

245 250 255

Gln Glu Leu Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile

260 265 270

Ala Gln Phe Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile

275 280 285

Gly Met Glu Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala

290 295 300

Ile Lys Leu Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu

305 310 315 320

Leu Asn Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro

325 330 335

Lys Lys Asp Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln

340 345 350

Arg Gln Lys Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu

355 360 365

Asp Asp Phe Ser Lys Gln Leu Gln Gln Ser Met Ala Ser Arg Val Gln

370 375 380

Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro

385 390 395 400

Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp

405 410 415

Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr

420 425 430

Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr

435 440 445

Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val

450 455 460

Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys

465 470 475 480

Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val

485 490 495

Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr

500 505 510

Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu

515 520 525

Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn

530 535 540

Gly Val Glu Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe

545 550 555 560

Gln Pro Thr Asn Gly Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu

565 570 575

Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys

580 585 590

Ser Thr Asn Leu Val Lys Asn Lys Ser Val Asn Phe

595 600

<210> 52

<211> 919

<212> PRT

<213> artificial sequence

<220>

<223> heavy chain fused to antigen

<400> 52

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ala Ser

435 440 445

Thr Asn Gly Ser Ile Thr Val Ala Ala Thr Ala Pro Thr Val Thr Pro

450 455 460

Thr Val Asn Ala Thr Pro Ser Ala Ala Ala Ser Arg Arg Gly Pro Glu

465 470 475 480

Gln Thr Gln Gly Asn Phe Gly Asp Gln Glu Leu Ile Arg Gln Gly Thr

485 490 495

Asp Tyr Lys His Trp Pro Gln Ile Ala Gln Phe Ala Pro Ser Ala Ser

500 505 510

Ala Phe Phe Gly Met Ser Arg Ile Gly Met Glu Val Thr Pro Ser Gly

515 520 525

Thr Trp Leu Thr Tyr Thr Gly Ala Ile Lys Leu Asp Asp Lys Asp Pro

530 535 540

Asn Phe Lys Asp Gln Val Ile Leu Leu Asn Lys His Ile Asp Ala Tyr

545 550 555 560

Lys Thr Phe Pro Pro Thr Glu Pro Lys Lys Asp Lys Lys Lys Lys Ala

565 570 575

Asp Glu Thr Gln Ala Leu Pro Gln Arg Gln Lys Lys Gln Gln Thr Val

580 585 590

Thr Leu Leu Pro Ala Ala Asp Leu Asp Asp Phe Ser Lys Gln Leu Gln

595 600 605

Gln Ser Met Ala Arg Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe

610 615 620

Ser Asn Asp Pro Phe Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser

625 630 635 640

Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr

645 650 655

Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln

660 665 670

Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly

675 680 685

Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp

690 695 700

Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile

705 710 715 720

Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser

725 730 735

Tyr Leu Thr Arg Pro Thr Ser Thr Pro Ala Asp Ser Ser Thr Ile Thr

740 745 750

Pro Thr Ala Thr Pro Thr Ala Thr Pro Thr Ile Lys Gly Ala Pro His

755 760 765

Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn

770 775 780

Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro

785 790 795 800

Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln

805 810 815

Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val

820 825 830

Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr

835 840 845

Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys

850 855 860

Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser

865 870 875 880

Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu

885 890 895

Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu

900 905 910

Leu Gly Lys Tyr Glu Gln Tyr

915

<210> 53

<211> 439

<212> PRT

<213> artificial sequence

<220>

<223> light chain fused to antigen

<400> 53

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys Ala Ser Arg Val Gln Pro Thr Glu Ser Ile

210 215 220

Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe

225 230 235 240

Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile

245 250 255

Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe

260 265 270

Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu

275 280 285

Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu

290 295 300

Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Lys Ile Ala Asp Tyr Asn

305 310 315 320

Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser

325 330 335

Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg

340 345 350

Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr

355 360 365

Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Glu Gly Phe

370 375 380

Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Asn Gly

385 390 395 400

Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu

405 410 415

His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val

420 425 430

Lys Asn Lys Ser Val Asn Phe

435

<210> 54

<211> 671

<212> PRT

<213> artificial sequence

<220>

<223> h anti-CD 40VH 3-LV-hIgGK-C-Virus SARS-CoV-2-spike-RBDC 221S variant

(south Africa mutation 20H/501Y.V2)

<400> 54

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ala Ser

435 440 445

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn

450 455 460

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

465 470 475 480

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

485 490 495

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

500 505 510

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

515 520 525

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

530 535 540

Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

545 550 555 560

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

565 570 575

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

580 585 590

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

595 600 605

Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

610 615 620

Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val

625 630 635 640

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

645 650 655

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Ser Val Asn Phe

660 665 670

<210> 55

<211> 687

<212> PRT

<213> artificial sequence

<220>

<223> h anti-CD 40VK2-LV-hIgGK-C-f 4-Virus SARS-Cov-2-Npep2-Spep1C12S-f2-Sp

ep4

<400> 55

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys Ala Ser Thr Asn Gly Ser Ile Thr Val Ala

210 215 220

Ala Thr Ala Pro Thr Val Thr Pro Thr Val Asn Ala Thr Pro Ser Ala

225 230 235 240

Ala Ala Ser Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp

245 250 255

Gln Glu Leu Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile

260 265 270

Ala Gln Phe Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile

275 280 285

Gly Met Glu Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala

290 295 300

Ile Lys Leu Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu

305 310 315 320

Leu Asn Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro

325 330 335

Lys Lys Asp Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln

340 345 350

Arg Gln Lys Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu

355 360 365

Asp Asp Phe Ser Lys Gln Leu Gln Gln Ser Met Ala Arg Asn Val Val

370 375 380

Ile Lys Val Cys Glu Phe Gln Phe Ser Asn Asp Pro Phe Leu Gly Val

385 390 395 400

Tyr Tyr His Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val

405 410 415

Tyr Ser Ser Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe

420 425 430

Leu Met Asp Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu

435 440 445

Phe Val Phe Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His

450 455 460

Thr Pro Ile Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu

465 470 475 480

Glu Pro Leu Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln

485 490 495

Thr Leu Leu Ala Leu His Arg Ser Tyr Leu Thr Arg Pro Thr Ser Thr

500 505 510

Pro Ala Asp Ser Ser Thr Ile Thr Pro Thr Ala Thr Pro Thr Ala Thr

515 520 525

Pro Thr Ile Lys Gly Ala Pro His Gly Val Val Phe Leu His Val Thr

530 535 540

Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys

545 550 555 560

His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn

565 570 575

Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile

580 585 590

Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile

595 600 605

Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp

610 615 620

Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro

625 630 635 640

Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn

645 650 655

Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn

660 665 670

Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr

675 680 685

<210> 56

<211> 611

<212> PRT

<213> artificial sequence

<220>

<223> H anti-CD 40VH3-LV-hIgG4H-C-f 4-Virus SARS-Cov-2-Npep2

<400> 56

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ala Ser

435 440 445

Thr Asn Gly Ser Ile Thr Val Ala Ala Thr Ala Pro Thr Val Thr Pro

450 455 460

Thr Val Asn Ala Thr Pro Ser Ala Ala Ala Ser Arg Arg Gly Pro Glu

465 470 475 480

Gln Thr Gln Gly Asn Phe Gly Asp Gln Glu Leu Ile Arg Gln Gly Thr

485 490 495

Asp Tyr Lys His Trp Pro Gln Ile Ala Gln Phe Ala Pro Ser Ala Ser

500 505 510

Ala Phe Phe Gly Met Ser Arg Ile Gly Met Glu Val Thr Pro Ser Gly

515 520 525

Thr Trp Leu Thr Tyr Thr Gly Ala Ile Lys Leu Asp Asp Lys Asp Pro

530 535 540

Asn Phe Lys Asp Gln Val Ile Leu Leu Asn Lys His Ile Asp Ala Tyr

545 550 555 560

Lys Thr Phe Pro Pro Thr Glu Pro Lys Lys Asp Lys Lys Lys Lys Ala

565 570 575

Asp Glu Thr Gln Ala Leu Pro Gln Arg Gln Lys Lys Gln Gln Thr Val

580 585 590

Thr Leu Leu Pro Ala Ala Asp Leu Asp Asp Phe Ser Lys Gln Leu Gln

595 600 605

Gln Ser Met

610

<210> 57

<211> 439

<212> PRT

<213> artificial sequence

<220>

<223> h anti-CD 40VK 2-LV-hIgGK-C-Virus SARS-CoV-2-spike-RBDC 221S SA var

<400> 57

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys Ala Ser Arg Val Gln Pro Thr Glu Ser Ile

210 215 220

Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe

225 230 235 240

Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile

245 250 255

Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe

260 265 270

Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu

275 280 285

Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu

290 295 300

Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala Asp Tyr Asn

305 310 315 320

Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser

325 330 335

Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg

340 345 350

Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr

355 360 365

Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Lys Gly Phe

370 375 380

Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Tyr Gly

385 390 395 400

Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu

405 410 415

His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val

420 425 430

Lys Asn Lys Ser Val Asn Phe

435

<210> 58

<211> 379

<212> PRT

<213> artificial sequence

<220>

<223> Multi-epitope polypeptide RBDSA VAR-Spep4

<400> 58

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn

1 5 10 15

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

20 25 30

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

35 40 45

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

50 55 60

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

65 70 75 80

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

85 90 95

Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

100 105 110

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

115 120 125

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

130 135 140

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

145 150 155 160

Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

165 170 175

Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val

180 185 190

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

195 200 205

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Ser Val Asn Phe Ala

210 215 220

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

225 230 235 240

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

245 250 255

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp

260 265 270

Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp

275 280 285

Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn

290 295 300

Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu

305 310 315 320

Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

325 330 335

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu

340 345 350

Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

355 360 365

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr

370 375

<210> 59

<211> 827

<212> PRT

<213> artificial sequence

<220>

<223> H anti-CD 40VH3-LV-hIgG 4H-C-Virus SARS-CoV-2-RBDC221S SA var-Spep4

<400> 59

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ala Ser

435 440 445

Arg Val Gln Pro Thr Glu Ser Ile Val Arg Phe Pro Asn Ile Thr Asn

450 455 460

Leu Cys Pro Phe Gly Glu Val Phe Asn Ala Thr Arg Phe Ala Ser Val

465 470 475 480

Tyr Ala Trp Asn Arg Lys Arg Ile Ser Asn Cys Val Ala Asp Tyr Ser

485 490 495

Val Leu Tyr Asn Ser Ala Ser Phe Ser Thr Phe Lys Cys Tyr Gly Val

500 505 510

Ser Pro Thr Lys Leu Asn Asp Leu Cys Phe Thr Asn Val Tyr Ala Asp

515 520 525

Ser Phe Val Ile Arg Gly Asp Glu Val Arg Gln Ile Ala Pro Gly Gln

530 535 540

Thr Gly Asn Ile Ala Asp Tyr Asn Tyr Lys Leu Pro Asp Asp Phe Thr

545 550 555 560

Gly Cys Val Ile Ala Trp Asn Ser Asn Asn Leu Asp Ser Lys Val Gly

565 570 575

Gly Asn Tyr Asn Tyr Leu Tyr Arg Leu Phe Arg Lys Ser Asn Leu Lys

580 585 590

Pro Phe Glu Arg Asp Ile Ser Thr Glu Ile Tyr Gln Ala Gly Ser Thr

595 600 605

Pro Cys Asn Gly Val Lys Gly Phe Asn Cys Tyr Phe Pro Leu Gln Ser

610 615 620

Tyr Gly Phe Gln Pro Thr Tyr Gly Val Gly Tyr Gln Pro Tyr Arg Val

625 630 635 640

Val Val Leu Ser Phe Glu Leu Leu His Ala Pro Ala Thr Val Cys Gly

645 650 655

Pro Lys Lys Ser Thr Asn Leu Val Lys Asn Lys Ser Val Asn Phe Ala

660 665 670

Ser Ala Pro His Gly Val Val Phe Leu His Val Thr Tyr Val Pro Ala

675 680 685

Gln Glu Lys Asn Phe Thr Thr Ala Pro Ala Ile Cys His Asp Gly Lys

690 695 700

Ala His Phe Pro Arg Glu Gly Val Phe Val Ser Asn Gly Thr His Trp

705 710 715 720

Phe Val Thr Gln Arg Asn Phe Tyr Glu Pro Gln Ile Ile Thr Thr Asp

725 730 735

Asn Thr Phe Val Ser Gly Asn Cys Asp Val Val Ile Gly Ile Val Asn

740 745 750

Asn Thr Val Tyr Asp Pro Leu Gln Pro Glu Leu Asp Ser Phe Lys Glu

755 760 765

Glu Leu Asp Lys Tyr Phe Lys Asn His Thr Ser Pro Asp Val Asp Leu

770 775 780

Gly Asp Ile Ser Gly Ile Asn Ala Ser Val Val Asn Ile Gln Lys Glu

785 790 795 800

Ile Asp Arg Leu Asn Glu Val Ala Lys Asn Leu Asn Glu Ser Leu Ile

805 810 815

Asp Leu Gln Glu Leu Gly Lys Tyr Glu Gln Tyr

820 825

<210> 60

<211> 379

<212> PRT

<213> artificial sequence

<220>

<223> h anti-CD 40VK2-LV-hIgGK-C-f 4-Virus SARS-CoV-2-Npep2

<400> 60

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys Ala Ser Thr Asn Gly Ser Ile Thr Val Ala

210 215 220

Ala Thr Ala Pro Thr Val Thr Pro Thr Val Asn Ala Thr Pro Ser Ala

225 230 235 240

Ala Ala Ser Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp

245 250 255

Gln Glu Leu Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile

260 265 270

Ala Gln Phe Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile

275 280 285

Gly Met Glu Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala

290 295 300

Ile Lys Leu Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu

305 310 315 320

Leu Asn Lys His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro

325 330 335

Lys Lys Asp Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln

340 345 350

Arg Gln Lys Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu

355 360 365

Asp Asp Phe Ser Lys Gln Leu Gln Gln Ser Met

370 375

<210> 61

<211> 263

<212> PRT

<213> artificial sequence

<220>

<223> Multi-epitope polypeptide Npep2-Spep1

<400> 61

Arg Arg Gly Pro Glu Gln Thr Gln Gly Asn Phe Gly Asp Gln Glu Leu

1 5 10 15

Ile Arg Gln Gly Thr Asp Tyr Lys His Trp Pro Gln Ile Ala Gln Phe

20 25 30

Ala Pro Ser Ala Ser Ala Phe Phe Gly Met Ser Arg Ile Gly Met Glu

35 40 45

Val Thr Pro Ser Gly Thr Trp Leu Thr Tyr Thr Gly Ala Ile Lys Leu

50 55 60

Asp Asp Lys Asp Pro Asn Phe Lys Asp Gln Val Ile Leu Leu Asn Lys

65 70 75 80

His Ile Asp Ala Tyr Lys Thr Phe Pro Pro Thr Glu Pro Lys Lys Asp

85 90 95

Lys Lys Lys Lys Ala Asp Glu Thr Gln Ala Leu Pro Gln Arg Gln Lys

100 105 110

Lys Gln Gln Thr Val Thr Leu Leu Pro Ala Ala Asp Leu Asp Asp Phe

115 120 125

Ser Lys Gln Leu Gln Gln Ser Met Ala Ser Asn Val Val Ile Lys Val

130 135 140

Cys Glu Phe Gln Phe Ser Asn Asp Pro Phe Leu Gly Val Tyr Tyr His

145 150 155 160

Lys Asn Asn Lys Ser Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser

165 170 175

Ala Asn Asn Cys Thr Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp

180 185 190

Leu Glu Gly Lys Gln Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe

195 200 205

Lys Asn Ile Asp Gly Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile

210 215 220

Asn Leu Val Arg Asp Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu

225 230 235 240

Val Asp Leu Pro Ile Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu

245 250 255

Ala Leu His Arg Ser Tyr Leu

260

<210> 62

<211> 738

<212> PRT

<213> artificial sequence

<220>

<223> H anti-CD 40VH3-LV-hIgG4H-C-f 4-Virus SARS-CoV-2-Npep2-Spep1C12S

<400> 62

Glu Val Gln Leu Val Glu Ser Gly Gly Gly Leu Val Gln Pro Gly Gly

1 5 10 15

Ser Leu Lys Leu Ser Cys Ala Thr Ser Gly Phe Thr Phe Ser Asp Tyr

20 25 30

Tyr Met Tyr Trp Val Arg Gln Ala Pro Gly Lys Gly Leu Glu Trp Val

35 40 45

Ala Tyr Ile Asn Ser Gly Gly Gly Ser Thr Tyr Tyr Pro Asp Thr Val

50 55 60

Lys Gly Arg Phe Thr Ile Ser Arg Asp Asn Ala Lys Asn Thr Leu Tyr

65 70 75 80

Leu Gln Met Asn Ser Leu Arg Ala Glu Asp Thr Ala Val Tyr Tyr Cys

85 90 95

Ala Arg Arg Gly Leu Pro Phe His Ala Met Asp Tyr Trp Gly Gln Gly

100 105 110

Thr Leu Val Thr Val Ser Ser Ala Ser Thr Lys Gly Pro Ser Val Phe

115 120 125

Pro Leu Ala Pro Cys Ser Arg Ser Thr Ser Glu Ser Thr Ala Ala Leu

130 135 140

Gly Cys Leu Val Lys Asp Tyr Phe Pro Glu Pro Val Thr Val Ser Trp

145 150 155 160

Asn Ser Gly Ala Leu Thr Ser Gly Val His Thr Phe Pro Ala Val Leu

165 170 175

Gln Ser Ser Gly Leu Tyr Ser Leu Ser Ser Val Val Thr Val Pro Ser

180 185 190

Ser Ser Leu Gly Thr Lys Thr Tyr Thr Cys Asn Val Asp His Lys Pro

195 200 205

Ser Asn Thr Lys Val Asp Lys Arg Val Glu Ser Lys Tyr Gly Pro Pro

210 215 220

Cys Pro Pro Cys Pro Ala Pro Glu Phe Glu Gly Gly Pro Ser Val Phe

225 230 235 240

Leu Phe Pro Pro Lys Pro Lys Asp Thr Leu Met Ile Ser Arg Thr Pro

245 250 255

Glu Val Thr Cys Val Val Val Asp Val Ser Gln Glu Asp Pro Glu Val

260 265 270

Gln Phe Asn Trp Tyr Val Asp Gly Val Glu Val His Asn Ala Lys Thr

275 280 285

Lys Pro Arg Glu Glu Gln Phe Asn Ser Thr Tyr Arg Val Val Ser Val

290 295 300

Leu Thr Val Leu His Gln Asp Trp Leu Asn Gly Lys Glu Tyr Lys Cys

305 310 315 320

Lys Val Ser Asn Lys Gly Leu Pro Ser Ser Ile Glu Lys Thr Ile Ser

325 330 335

Lys Ala Lys Gly Gln Pro Arg Glu Pro Gln Val Tyr Thr Leu Pro Pro

340 345 350

Ser Gln Glu Glu Met Thr Lys Asn Gln Val Ser Leu Thr Cys Leu Val

355 360 365

Lys Gly Phe Tyr Pro Ser Asp Ile Ala Val Glu Trp Glu Ser Asn Gly

370 375 380

Gln Pro Glu Asn Asn Tyr Lys Thr Thr Pro Pro Val Leu Asp Ser Asp

385 390 395 400

Gly Ser Phe Phe Leu Tyr Ser Arg Leu Thr Val Asp Lys Ser Arg Trp

405 410 415

Gln Glu Gly Asn Val Phe Ser Cys Ser Val Met His Glu Ala Leu His

420 425 430

Asn His Tyr Thr Gln Lys Ser Leu Ser Leu Ser Leu Gly Lys Ala Ser

435 440 445

Thr Asn Gly Ser Ile Thr Val Ala Ala Thr Ala Pro Thr Val Thr Pro

450 455 460

Thr Val Asn Ala Thr Pro Ser Ala Ala Ala Ser Arg Arg Gly Pro Glu

465 470 475 480

Gln Thr Gln Gly Asn Phe Gly Asp Gln Glu Leu Ile Arg Gln Gly Thr

485 490 495

Asp Tyr Lys His Trp Pro Gln Ile Ala Gln Phe Ala Pro Ser Ala Ser

500 505 510

Ala Phe Phe Gly Met Ser Arg Ile Gly Met Glu Val Thr Pro Ser Gly

515 520 525

Thr Trp Leu Thr Tyr Thr Gly Ala Ile Lys Leu Asp Asp Lys Asp Pro

530 535 540

Asn Phe Lys Asp Gln Val Ile Leu Leu Asn Lys His Ile Asp Ala Tyr

545 550 555 560

Lys Thr Phe Pro Pro Thr Glu Pro Lys Lys Asp Lys Lys Lys Lys Ala

565 570 575

Asp Glu Thr Gln Ala Leu Pro Gln Arg Gln Lys Lys Gln Gln Thr Val

580 585 590

Thr Leu Leu Pro Ala Ala Asp Leu Asp Asp Phe Ser Lys Gln Leu Gln

595 600 605

Gln Ser Met Ala Ser Asn Val Val Ile Lys Val Cys Glu Phe Gln Phe

610 615 620

Ser Asn Asp Pro Phe Leu Gly Val Tyr Tyr His Lys Asn Asn Lys Ser

625 630 635 640

Trp Met Glu Ser Glu Phe Arg Val Tyr Ser Ser Ala Asn Asn Cys Thr

645 650 655

Phe Glu Tyr Val Ser Gln Pro Phe Leu Met Asp Leu Glu Gly Lys Gln

660 665 670

Gly Asn Phe Lys Asn Leu Arg Glu Phe Val Phe Lys Asn Ile Asp Gly

675 680 685

Tyr Phe Lys Ile Tyr Ser Lys His Thr Pro Ile Asn Leu Val Arg Asp

690 695 700

Leu Pro Gln Gly Phe Ser Ala Leu Glu Pro Leu Val Asp Leu Pro Ile

705 710 715 720

Gly Ile Asn Ile Thr Arg Phe Gln Thr Leu Leu Ala Leu His Arg Ser

725 730 735

Tyr Leu

<210> 63

<211> 595

<212> PRT

<213> artificial sequence

<220>

<223> h anti-CD 40VK 2-LV-hIgGK-C-Virus SARS-CoV-2-RBDC221S SA var-Spep4

<400> 63

Asp Ile Gln Met Thr Gln Ser Pro Ser Ser Leu Ser Ala Ser Val Gly

1 5 10 15

Asp Arg Val Thr Ile Thr Cys Ser Ala Ser Gln Gly Ile Ser Asn Tyr

20 25 30

Leu Asn Trp Tyr Gln Gln Lys Pro Gly Lys Ala Val Lys Leu Leu Ile

35 40 45

Tyr Tyr Thr Ser Ile Leu His Ser Gly Val Pro Ser Arg Phe Ser Gly

50 55 60

Ser Gly Ser Gly Thr Asp Tyr Thr Leu Thr Ile Ser Ser Leu Gln Pro

65 70 75 80

Glu Asp Phe Ala Thr Tyr Tyr Cys Gln Gln Phe Asn Lys Leu Pro Pro

85 90 95

Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg Thr Val Ala Ala

100 105 110

Pro Ser Val Phe Ile Phe Pro Pro Ser Asp Glu Gln Leu Lys Ser Gly

115 120 125

Thr Ala Ser Val Val Cys Leu Leu Asn Asn Phe Tyr Pro Arg Glu Ala

130 135 140

Lys Val Gln Trp Lys Val Asp Asn Ala Leu Gln Ser Gly Asn Ser Gln

145 150 155 160

Glu Ser Val Thr Glu Gln Asp Ser Lys Asp Ser Thr Tyr Ser Leu Ser

165 170 175

Ser Thr Leu Thr Leu Ser Lys Ala Asp Tyr Glu Lys His Lys Val Tyr

180 185 190

Ala Cys Glu Val Thr His Gln Gly Leu Ser Ser Pro Val Thr Lys Ser

195 200 205

Phe Asn Arg Gly Glu Cys Ala Ser Arg Val Gln Pro Thr Glu Ser Ile

210 215 220

Val Arg Phe Pro Asn Ile Thr Asn Leu Cys Pro Phe Gly Glu Val Phe

225 230 235 240

Asn Ala Thr Arg Phe Ala Ser Val Tyr Ala Trp Asn Arg Lys Arg Ile

245 250 255

Ser Asn Cys Val Ala Asp Tyr Ser Val Leu Tyr Asn Ser Ala Ser Phe

260 265 270

Ser Thr Phe Lys Cys Tyr Gly Val Ser Pro Thr Lys Leu Asn Asp Leu

275 280 285

Cys Phe Thr Asn Val Tyr Ala Asp Ser Phe Val Ile Arg Gly Asp Glu

290 295 300

Val Arg Gln Ile Ala Pro Gly Gln Thr Gly Asn Ile Ala Asp Tyr Asn

305 310 315 320

Tyr Lys Leu Pro Asp Asp Phe Thr Gly Cys Val Ile Ala Trp Asn Ser

325 330 335

Asn Asn Leu Asp Ser Lys Val Gly Gly Asn Tyr Asn Tyr Leu Tyr Arg

340 345 350

Leu Phe Arg Lys Ser Asn Leu Lys Pro Phe Glu Arg Asp Ile Ser Thr

355 360 365

Glu Ile Tyr Gln Ala Gly Ser Thr Pro Cys Asn Gly Val Lys Gly Phe

370 375 380

Asn Cys Tyr Phe Pro Leu Gln Ser Tyr Gly Phe Gln Pro Thr Tyr Gly

385 390 395 400

Val Gly Tyr Gln Pro Tyr Arg Val Val Val Leu Ser Phe Glu Leu Leu

405 410 415

His Ala Pro Ala Thr Val Cys Gly Pro Lys Lys Ser Thr Asn Leu Val

420 425 430

Lys Asn Lys Ser Val Asn Phe Ala Ser Ala Pro His Gly Val Val Phe

435 440 445

Leu His Val Thr Tyr Val Pro Ala Gln Glu Lys Asn Phe Thr Thr Ala

450 455 460

Pro Ala Ile Cys His Asp Gly Lys Ala His Phe Pro Arg Glu Gly Val

465 470 475 480

Phe Val Ser Asn Gly Thr His Trp Phe Val Thr Gln Arg Asn Phe Tyr

485 490 495

Glu Pro Gln Ile Ile Thr Thr Asp Asn Thr Phe Val Ser Gly Asn Cys

500 505 510

Asp Val Val Ile Gly Ile Val Asn Asn Thr Val Tyr Asp Pro Leu Gln

515 520 525

Pro Glu Leu Asp Ser Phe Lys Glu Glu Leu Asp Lys Tyr Phe Lys Asn

530 535 540

His Thr Ser Pro Asp Val Asp Leu Gly Asp Ile Ser Gly Ile Asn Ala

545 550 555 560

Ser Val Val Asn Ile Gln Lys Glu Ile Asp Arg Leu Asn Glu Val Ala

565 570 575

Lys Asn Leu Asn Glu Ser Leu Ile Asp Leu Gln Glu Leu Gly Lys Tyr

580 585 590

Glu Gln Tyr

595

Claims

1. A SARS-CoV-2 polypeptide (N276-411, "Npep 2") that is derived from protein N and consists of a polypeptide that hybridizes with SEQ ID NO:2 from residue 276 to residue 411, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

2. The polypeptide (N276-411) of claim 1, comprising a sequence identical to SEQ ID NO:2 from residue 276 to residue 411; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135 or 136 consecutive amino acids.

3. The polypeptide (N276-411) of claim 1, which consists of a polypeptide sequence identical to SEQ ID NO:2 from residue 276 to residue 411.

4. A SARS-CoV-2 polypeptide (S125-250, "Spep 1") that is derived from protein S and consists of a polypeptide that hybridizes to SEQ ID NO:3 from residue 125 to residue 250, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

5. The polypeptide (S125-250) of claim 4, comprising a sequence identical to SEQ ID NO:3 from residue 125 to residue 250, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125 or 126 consecutive amino acids.

6. The polypeptide (S125-250) according to claim 4, consisting of a polypeptide sequence identical to SEQ ID NO:3 from residue 125 to residue 250.

7. The polypeptide (S125-250) according to claim 4, consisting of an amino acid sequence in SEQ ID NO:3 to residue at position 125 and to residue at position 250, and comprises a C136S non-naturally occurring mutation.

8. A SARS-CoV-2 polypeptide (S1056-1209, "Spep 4") that is derived from protein S and consists of a polypeptide that hybridizes to SEQ ID NO:3 from residue 1056 to residue 1209, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

9. The polypeptide (S1056-1209) of claim 8, comprising an amino acid sequence that hybridizes to SEQ ID NO:3 from residue 1056 to residue 1209, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135;136;137, respectively; 138;139, respectively; 140;141;142;143, a base; 144 (144); 145, respectively; 146;147;148, a step of selecting a key; 149;150;151;152;153 or 154 consecutive amino acids.

10. The polypeptide (S1056-1209) according to claim 9, which consists of a polypeptide sequence that hybridizes with SEQ ID NO:3 from residue 1056 to residue 1209.

11. A SARS-CoV-2 polypeptide (M1-110) that is derived from protein M and consists of a polypeptide that hybridizes to SEQ ID NO:1 from residue 1 to residue 110, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

12. The polypeptide (M1-110) according to claim 11, comprising a sequence identical to SEQ ID NO:1 from residue 1 to residue 110, 50 of the amino acid sequences having at least 90% identity to the amino acid sequence of seq id no; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109 or 110 consecutive amino acids.

13. The polypeptide (M1-110) according to claim 11, consisting of a polypeptide sequence identical to SEQ ID NO:1 from residue 1 to residue 110.

14. A SARS-CoV-2 polypeptide (M132-222) that is derived from protein M and consists of a polypeptide that hybridizes to SEQ ID NO:1 from residue 132 to residue 222, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

15. The polypeptide (M132-222) of claim 14, comprising a sequence that hybridizes to SEQ ID NO:1 from residue 132 to residue 222, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90 or 91 consecutive amino acids.

16. The polypeptide (M132-222) of claim 14, which consists of a polypeptide sequence that hybridizes to SEQ ID NO:1 from residue 132 to residue 222, and an amino acid sequence having at least 90% identity.

17. A SARS-CoV-2 polypeptide (N78-206) that is derived from protein N and consists of a polypeptide that is identical to SEQ ID NO:2 from residue 78 to residue 206, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity to the amino acid sequence.

18. The polypeptide (N78-206) according to claim 17, comprising a sequence identical to SEQ ID NO:2 from residue 78 to residue 206, 50 of the amino acid sequences having at least 90% identity to the amino acid sequence of seq id no; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127;128 or 129 consecutive amino acids.

19. The polypeptide (N78-206) according to claim 17, which consists of a polypeptide sequence identical to SEQ ID NO:2 from residue 78 to residue 206.

20. A SARS-CoV-2 polypeptide (S280-598) that is derived from protein S and consists of a polypeptide that hybridizes with SEQ ID NO:3 from residue 280 to residue 598, and at least 50 consecutive amino acids in the amino acid sequence having at least 90% identity.

21. The polypeptide (S280-598) of claim 20, comprising a sequence that hybridizes to SEQ ID NO:3 from residue 280 to residue 598, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135;136;137, respectively; 138;139, respectively; 140;141;142;143, a base; 144 (144); 145, respectively; 146;147;148, a step of selecting a key; 149;150;151;152;153;154;155;156, respectively; 157 (157); 158;159;160, a step of detecting a position of the base; 161;162;163;164, a step of detecting the position of the base; 165;166;167;168;169 (169); 170, a step of; 171;172;173, a part of the base; 174;175;176;177 (step 1); 178;179;180;181;182;183 (V); 184;185;186, a step of detecting the position of the base; 187;188;189;190;191; 192. 193;194, a step of receiving a signal; 195;196;197;198, a step of; 199;200;201;202;203, a base station; 204; 205. 206;207 (x, y); 208;209;210;211;212;213;214;215, respectively; 216;217;218;219, a step of; 220;221;222, a step of; 223, a third step; 224;225;226;227;228;229;230, a step of; 231;232;233;234;235;236;237; 238. 239;240, a step of; 241, a base; 242;243;244;245; 246. 247, respectively; 248;249;250;251; 252. 253;254, a base plate; 255, respectively; 256;257;258;259;260;261;262;263;264;265;266;267;268, a step of; 269;270; 271(s); 272;273;274;275;276, respectively; 277;278;279;280; 281. 282;283;284;285;286;287;288;289;290;291 of a metal strip; 292;293;294;295;296;297;298;299;300;301;302;303;304;305; 306. 307. 308. 309, and (c) a third party; 310;311;312;313;314, a step of; 315;316, a step of; 317. 318 or 319 consecutive amino acids.

22. The polypeptide (S280-598) of claim 20, which consists of a polypeptide sequence that hybridizes with SEQ ID NO:3 from residue 280 to residue 598.

23. The polypeptide (S280-598) according to claim 20, consisting of an amino acid sequence in SEQ ID NO:3 from residue 280 to residue 598 and comprises one or more naturally occurring mutations selected from the group consisting of the K417N, K417T, E K and N501Y mutations.

24. The polypeptide (S280-598) according to claim 20, consisting of an amino acid sequence in SEQ ID NO:3 to residue position 598, and comprises a naturally occurring mutation of K417N, E484K, N501Y and a non-naturally occurring mutation of C538S.

25. A SARS-CoV-2 polypeptide (S680-1029) that is derived from protein S and consists of a polypeptide that hybridizes to SEQ ID NO:3 from residue 680 to residue 1029, and at least 50 consecutive amino acids in an amino acid sequence having at least 90% identity.

26. The polypeptide (S680-1029) of claim 25, comprising a sequence identical to SEQ ID NO:3 from residue 680 to residue 1029, 50 of the amino acid sequences having at least 90% identity; 51;52;53;54;55;56;57;58;59;60;61;62;63;64; 65. 66;67;68;69;70;71;72;73;74;75;76;77;78;79;80;81;82;83, a step of detecting the position of the base; 84;85;86;87, a base; 88;89;90;91;92;93;94;95;96;97;98;99, a step of; 100;101;102, a step of; 103;104;105; 106. 107;108, a step of; 109;110;111;112;113; 114. 115;116;117;118;119;120;121;122, a step of; 123, a step of; 124;125;126;127; 128. 129, respectively; 130;131;132, a part of the material; 133;134;135;136;137, respectively; 138;139, respectively; 140;141;142;143, a base; 144 (144); 145, respectively; 146;147;148, a step of selecting a key; 149;150;151;152;153;154;155;156, respectively; 157 (157); 158;159;160, a step of detecting a position of the base; 161;162;163;164, a step of detecting the position of the base; 165;166;167;168;169 (169); 170, a step of; 171;172;173, a part of the base; 174;175;176;177 (step 1); 178;179;180;181;182;183 (V); 184;185;186, a step of detecting the position of the base; 187;188;189;190;191; 192. 193;194, a step of receiving a signal; 195;196;197;198, a step of; 199;200;201;202;203, a base station; 204; 205. 206;207 (x, y); 208;209;210;211;212;213;214;215, respectively; 216;217;218;219, a step of; 220;221;222, a step of; 223, a third step; 224;225;226;227;228;229;230, a step of; 231;232;233;234;235;236;237; 238. 239;240, a step of; 241, a base; 242;243;244;245; 246. 247, respectively; 248;249;250;251; 252. 253;254, a base plate; 255, respectively; 256;257;258;259;260;261;262;263;264;265;266;267;268, a step of; 269;270; 271(s); 272;273;274;275;276, respectively; 277;278;279;280; 281. 282;283;284;285;286;287;288;289;290;291 of a metal strip; 292;293;294;295;296;297;298;299;300;301;302;303;304;305; 306. 307. 308. 309, and (c) a third party; 310;311;312;313;314, a step of; 315;316, a step of; 317. 318;319; 320. 321, a base; 322;323 (323); 324, a base; 325, a step of; 326, a step of; 327, respectively; 328. 329;330;331;332;333;334;335;336, a base; 337, respectively; 338;339;340 (340); 341;342;343;344;345, a frame structure; 346;347;348;349 or 350 consecutive amino acids.

27. The polypeptide (S680-1029) of claim 25, which consists of a polypeptide sequence identical to SEQ ID NO:3 from residue 680 to residue 1029.

28. A conjugate, wherein a heterologous polypeptide is conjugated or fused to one or more SARS-CoV-2 polypeptides of any one of claims 1-27.

29. The conjugate of claim 28, comprising the polypeptide of claim 1 (N276-411, npep 2).

30. The conjugate of claim 29, comprising a polypeptide (N276-411 "npep 2"), a polypeptide (S125-250, "Spep 1"), and a polypeptide (S1056-1209, "Spep 4").

31. The conjugate of claim 30, comprising a polypeptide having the amino acid sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

32. The conjugate of claim 29, comprising a polypeptide (S125-250, "Spep 1") and a polypeptide (S1056-1209, "Spep 4").

33. The conjugate of claim 32, comprising a polypeptide having the amino acid sequence as set forth in SEQ ID NO:6, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

34. The conjugate of claim 29, comprising a polypeptide (N276-411, "Npep 2") and an RBD polypeptide.

35. The conjugate of claim 34, comprising a polypeptide having the amino acid sequence as set forth in SEQ ID NO: 7.

36. The conjugate of claim 29, wherein the heterologous polypeptide is an immunoglobulin domain, in particular a heavy chain or a light chain of an antibody.

37. The conjugate of claim 36, wherein the antibody is an IgG antibody, preferably an IgG1 or IgG4 antibody, or even more preferably an IgG4 antibody.

38. The conjugate of claim 36, wherein the antibody is directed against a surface antigen of an APC selected from the group consisting of: DC immunoreceptor (DCIR), MHC class I, MHC class II, CD1, CD2, CD3, CD4, CD8, CD11B, CD14, CD15, CD16, CD19, CD20, CD29, CD31, CD40, CD43, CD44, CD45, CD54, CD56, CD57, CD58, CD83, CD86, CMRF-44, CMRF-56, DCIR, DC-ASPGR, CLEC-6, CD40, BDCA-2, MARCO, DEC-205, mannose receptor, langerhans's, DECTIN-1, B7-2, IFN-gamma receptor and IL-2 receptor, ICAM-1, fey receptor, LOX-1 and ASPGR.

39. The conjugate of claim 38, wherein the antibody is directed against CD40.

40. The conjugate of claim 39, wherein the CD40 antibody is derived from

-12E12 antibody, and which comprises:

a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H, said CDR1H having amino acid sequence GFTFSDYYMY (SEQ ID NO: 8), said CDR2H having amino acid sequence YINSGGGSTYYPDTVKG (SEQ ID NO: 9) and said CDR3H having amino acid sequence RGLPFHAMDY (SEQ ID NO: 10);

and a light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L, said CDR1L having amino acid sequence SASQGISNYLN (SEQ ID NO: 11), said CDR2L having amino acid sequence YTILS (SEQ ID NO: 12) and said CDR3L having amino acid sequence QQFNKLPPT (SEQ ID NO: 13), or

-11B6 antibody, and which comprises:

a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H, said CDR1H having amino acid sequence GYSFTGYYMH (SEQ ID NO: 14), said CDR2H having amino acid sequence RINPYNGATSYNQNFKD (SEQ ID NO: 15), said CDR3H having amino acid sequence EDYVY (SEQ ID NO: 16), and

a light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L, said CDR1L having amino acid sequence RSSQSLVHSNGNTYLH (SEQ ID NO: 17), said CDR2L having amino acid sequence KVSNRFS (SEQ ID NO: 18) and said CDR3L having amino acid sequence SQSTHVPWT (SEQ ID NO: 19), or

-12B4 antibody, and which comprises:

a heavy chain comprising complementarity determining regions CDR1H, CDR H and CDR3H, said CDR1H having amino acid sequence GYTFTDYVLH (SEQ ID NO: 20), said CDR2H having amino acid sequence YINPYNDGTKYNEKFKG (SEQ ID NO: 21), said CDR3H having amino acid sequence GYPAYSGYAMDY (SEQ ID NO: 22), and

light chain comprising complementarity determining regions CDR1L, CDR L and CDR3L, said CDR1L having amino acid sequence RASQDISNYLN (SEQ ID NO: 23), said CDR2L having amino acid sequence YTSRLHS (SEQ ID NO: 24) and said CDR3L having amino acid sequence HHGNTLPWT (SEQ ID NO: 25).

41. The conjugate of claim 39, wherein the CD40 antibody is selected from mAb1, mAb2, mAb3, mAb4, mAb5, and mAb6 selected as set forth in table a.

42. The conjugate of claim 38, wherein the antibody is specific for Yu Langge hanstin.

43. The conjugate of claim 42, wherein the antibody is derived from antibody 15B10 having ATCC accession No. PTA-9852, or from antibody 2G3 having ATCC accession No. PTA-9853, or from antibody 91E7, 37C1 or 4C7 described in WO 2011032161.

44. The conjugate according to claim 43, wherein the antibody is selected from the group consisting of mAb7, mAb8, mAb9, mAb10, mAb11 and mAb12 selected as set forth in table B.

45. The conjugate of claim 28, wherein the heterologous polypeptide is fused to one or more SARS-CoV-2 polypeptides of any one of claims 1 to 27 to form a fusion protein.

46. The conjugate according to claim 45, wherein the SARS-CoV-2 polypeptide is fused to the heterologous polypeptide either directly or through a linker.

47. The conjugate according to claim 46, wherein the linker is selected from FlexV1, f2, f3 or f4.

48. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "Gen2 a"), wherein:

-the heavy chain of said antibody is fused to an RDB polypeptide, and

49. The conjugate according to claim 48, wherein the light chain of the Gen2a antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

50. The conjugate according to claim 48, wherein said Gen2a antibody comprises the amino acid sequence set forth in SEQ ID NO:48, and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:49, and a light chain of the amino acid sequence shown in seq id no.

51. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "Gen2 b"), wherein:

-the heavy chain of the antibody is fused to a multi-epitope polypeptide comprising a polypeptide (S125-250, "Spep 1") and a polypeptide (S1056-1209, "Spep 4");

52. The conjugate of claim 51, wherein the heavy chain of the Gen2b antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:6, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

53. The conjugate of claim 51, wherein the light chain of Gen2b is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO: 7.

54. The conjugate of claim 51, wherein the Gen2b antibody comprises the amino acid sequence set forth in SEQ ID NO:50, and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:51, and a light chain of the amino acid sequence shown in seq id no.

55. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "Gen2 c"), wherein:

the heavy chain of said antibody is fused to a multi-epitope polypeptide comprising a polypeptide (N276-411 "Npep 2"), a polypeptide (S125-250, "Spep 1") and a polypeptide (S1056-1209, "Spep 4"), and

-the light chain of the antibody is fused to an RDB polypeptide.

56. The conjugate of claim 55, wherein the heavy chain of the Gen2c antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

57. The conjugate of claim 55, wherein the Gen2c antibody comprises the amino acid sequence set forth in SEQ ID NO:52, and having the heavy chain as set forth in SEQ ID NO:53, and a light chain of the amino acid sequence shown in seq id no.

58. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "CD40.Cov2 v"), wherein:

-the heavy chain of said antibody is fused to an RDB polypeptide, and

59. The conjugate according to claim 58, wherein the light chain of the cd40.cov2 antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:5, and a multi-epitope polypeptide of the amino acid sequence shown in the specification.

60. The conjugate according to claim 58, comprising the amino acid sequence set forth in SEQ ID NO:54, and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:55, and a light chain of the amino acid sequence shown in seq id no.

61. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "CD40.N2. Rbdv"), wherein:

The heavy chain of said antibody is fused to a polypeptide (N276-411 "Npep 2"),

-the light chain of the antibody is fused to an RBD polypeptide.

62. The conjugate according to claim 61, comprising the amino acid sequence set forth in SEQ ID NO:56, and a heavy chain having the amino acid sequence as set forth in SEQ ID NO: 57.

63. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "CD40.N2. Rbdv-2"), wherein:

the heavy chain of said antibody is fused to an RBD polypeptide,

the light chain of said antibody is fused to a polypeptide (N276-411 "Npep 2").

64. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "CD40. Rbdv.s4.n2"), wherein:

the heavy chain of said antibody is fused to a multi-epitope polypeptide comprising an RBD polypeptide and a polypeptide (S1056-1209, "Spep 4"),

the light chain of the antibody is fused to a polypeptide (N276-411 "Npep 2").

65. The conjugate according to claim 64, wherein the heavy chain of the cd40.rbdv.s4.n2 antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:58, and a multi-epitope polypeptide of the amino acid sequence shown in seq id no.

66. The conjugate according to claim 64, comprising the amino acid sequence set forth in SEQ ID NO:59, and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:60, and a light chain of the amino acid sequence shown in seq id no.

67. The conjugate according to any one of claims 39 to 41, consisting of a CD40 antibody (designated "CD40.N2s1.Rbdv s 4"), wherein:

the heavy chain of said antibody is fused to a multi-epitope polypeptide comprising a polypeptide (N276-411 "Npep 2") and a polypeptide (S125-250, "Spep 1"),

68. The conjugate according to claim 67, wherein the light chain of the cd40.n2s1.rbdvs4 antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:58, and a multi-epitope polypeptide of the amino acid sequence shown in seq id no.

69. The conjugate according to claim 67, wherein the heavy chain of the cd40.n2s1.rbdvs4 antibody is fused to a polypeptide having the amino acid sequence as set forth in SEQ ID NO:61, and a polypeptide having the amino acid sequence shown in seq id no.

70. The conjugate according to claim 67, comprising the amino acid sequence set forth in SEQ ID NO:62, and a heavy chain having the amino acid sequence as set forth in SEQ ID NO:63, and a light chain of the amino acid sequence shown in seq id no.

71. The conjugate of claim 58, 61, 63, 64, or 67, wherein the RBD polypeptide consists of amino acids that are set forth in SEQ ID NO:3 from amino acid residue 319 to amino acid residue 541 and comprises a naturally occurring mutation of K417N, E484K, N Y501Y and a non-naturally occurring C538S mutation ("RBD" _{South Africa variants} ”)。

72. A vaccine composition comprising one or more SARS-CoV-2 polypeptide according to any one of claims 1 to 27 as antigen and/or the conjugate according to any one of claims 28 to 71.

73. The vaccine composition of claim 72, comprising 2, 3, 4, 5, 6, 7 or 8 SARS-CoV-2 polypeptides of any one of claims 1-27.

74. A polynucleotide encoding one or more SARS-CoV-2 polypeptides according to any one of claims 1 to 27 and/or the conjugate according to any one of claims 28 to 71.

75. A method of vaccinating a subject in need thereof against SARS-CoV-2 comprising administering a therapeutically effective amount of one or more SARS-CoV-2 polypeptides of any one of claims 1 to 27, and/or one or more conjugates of any one of claims 28 to 71, and/or the vaccine composition of claim 72, and/or the polynucleotide of claim 74.