Hepatitis B virus vaccine

Abstract

The present disclosure relates to isolated polynucleotides and polypeptides, and related hepatitis B virus (HBV) vaccines. The present disclosure also relates to viral vectors for expressing such polypeptides and for use in HBV vaccines, as well as methods for protecting subjects from HBV infection and methods for treating HBV in subjects, the methods comprising administering polypeptides, vectors or vaccines described herein. Also provided herein is a method for designing and producing HBV vaccines, the methods comprising designing vaccine antigens using antigen amino acid sequences to cover diversity within a geographic region, the antigen amino acid sequences effectively covering HBV genotype epitopes present in the geographic region.

Description

[0001] Government Interest Statement

[0002] The United States government has certain rights to this invention under contract number 89233218CNA000001 between the U.S. Department of Energy and TRIAD National Security, LLC regarding the operation of Los Alamos National Laboratory.

[0003] Declaration of sequence list

[0004] The sequence listings related to this application are provided in text format in lieu of a paper copy and are incorporated herein by reference. The name of the text file containing the sequence listings is 930185_414WO_SEQUENCE_LISTING.txt. The text file is 186KB in size, created on August 21, 2020, and submitted electronically via EFS-Web. Technical Field

[0005] This topic typically involves hepatitis B virus (HBV), particularly HBV vaccines. Background Technology

[0006] Hepatitis B virus (HBV) infection represents a major global health burden. HBV infection causes 5–10% of infected adults to develop chronic liver disease, while the rate is reversed for perinatal transmission, with >90% of these cases progressing to chronic disease. Untreated chronic hepatitis B (CHB) infection often progresses to necrotizing inflammation and persistent liver damage, leading to cirrhosis and hepatocellular carcinoma (HCC). CHB is estimated to increase the risk of HCC by 20 times and accounts for approximately 54% of HCC cases. HCC is the third leading cause of cancer death, with approximately 800,000 new cases diagnosed each year. Highly effective (>95%) prophylactic vaccines were introduced in the early 1980s; however, these HBV vaccines become ineffective once infection is confirmed. Despite widespread use of prophylactic vaccines, there are still 240–340 million chronic HBV carriers and more than 780,000 related deaths worldwide each year. The vast majority of new HBV infections occur in high-prevalence areas such as China, Southeast Asia, and sub-Saharan Africa. HBV infection can be transmitted through sexual contact, hospitals, or blood.

[0007] The HBV genome is a 3.2kB double-stranded DNA molecule organized into four overlapping open reading frames: polymerase (P or Pol), core (C), surface antigen (HBsAg) (S), and a gene called “X” (whose function is not fully understood but is associated with the progression of liver cancer). (Locarnini et al. Antivir Ther. 2010; 15 Supplement 3: 3-14). Moreover, unlike other small chronic viruses (such as HIV or HCV) that exhibit high mutation rates leading to quasi-species, HBV’s small (3.2kb) DNA genome is more limited in its mutational capacity due to the overlapping open reading frames (ORFs). Although intrahost mutations are limited, different HBV genotypes have been identified globally, and these genotypes need to be considered when designing therapeutics (e.g., vaccines).

[0008] The current standard of care for chronic HBV infection is treatment with antiviral drugs and interferon-alpha.

[0009] To date, therapeutic vaccination against HBV has been ineffective. There are many potential reasons for this. Identifying the HBV sequence in infected individuals helps determine which specific vaccines will provide effective treatment. The HBV database, known as “HBVdb,” developed by a collaborative consortium that sequences and examines HBV genomes, has identified approximately 5,000 complete HBV genomes. See Hayer et al. Nucleic Acids Res., 41:D566-D570 2013, the entire contents of which are incorporated herein by reference. However, due to widespread sequence overlap and because multiple sequences are isolated from the same patient or from highly compact clusters, the total number of truly unique complete HBV genomes identified by HBVdb is roughly 3,000. The HBV genomes identified by HBVdb can be classified into different genotypes (A, B, C, CB, D, DC, DE, E, and F). The distribution of genotypes varies across the world's population. For example, in samples from individuals from Asia, particularly China, nearly 75% of the sequences available on HBVdb contain genotypes B and C. On the other hand, more than half of HBV infections in Europe represent infections of genotypes A and D.

[0010] While antiviral drugs and interferon-alpha treatment suppress viral replication and stimulate the innate immune system, they rarely clear the virus (1-4% per year), and patients typically require lifelong treatment. Furthermore, individual responses to these therapies vary as the disease progresses, and long-term treatment can lead to both drug resistance mutations and a wide range of side effects. The most effective direct-acting antiviral therapies, tenofovir and entecavir, are expensive and only available to a subset of patients using expert guidelines and algorithms. With treatment failure or discontinuation, liver inflammation becomes chronic, and cycles of damage and regeneration can lead to fibrosis, abnormal liver structure, and potentially hepatocellular carcinoma (HCC). Therefore, there is an urgent need for an effective immunotherapy that progressively increases the effectiveness of the immune response, leading to HBV elimination and eventual cure.

[0011] A small percentage of individuals with chronic HBV will spontaneously clear their infection, as evidenced by the loss of surface antigen measured in the blood. This rate is as low as 1% per year, and minimal improvement can be achieved with the use of direct-acting antiviral agents. The most effective clearance occurs in individuals with low circulating surface antigen levels who are receiving type 1 interferon therapy, which has serious side effects and is generally poorly tolerated.

[0012] Therapeutic vaccination is also partially ineffective because the T cells required to clear the infection are depleted or resistant, and do not lead to effective clearance of HBV from the liver. Therefore, it is necessary to elicit T cells that are not induced by natural infection but can still recognize peptide sequences displayed on the surface of infected hepatocytes.

[0013] CMV / HBV vaccines offer a pathway to gradually increase an effective response leading to HBV elimination and eventual cure. Induction of an active hepatic-resident effector CD8+ T cell (TEM) response is crucial for HBV clearance. Therefore, sustained induction of HBV-specific effector CD8+ T cells that recognize novel epitopes in the liver of CHB patients should be sufficient to control and ultimately eliminate HBV.

[0014] Therefore, there remains a need in the art to confirm effective vaccination for people infected with different HBV genotypes. Furthermore, since currently available vaccines against chronic HBV infection are ineffective, there is a need in the art to develop a vaccine for treating such infections. There is also a need to design, prepare, and test therapeutic HBV vaccines in preparation for clinical trials. The compositions and methods disclosed herein address these needs. Summary of the Invention

[0015] In some aspects, this disclosure provides polypeptides comprising amino acid sequences as listed in SEQ ID NO:1-11 or SEQ ID NO:14-36. In some embodiments, the polypeptide comprises two or more amino acid sequences as listed in SEQ ID NO:16-36. In some embodiments, the above-described polypeptide, or the polynucleotide encoding the polypeptide, can be used in an HBV vaccine. In some embodiments, this disclosure provides immunogenic compositions comprising polypeptides comprising amino acid sequences as listed in SEQ ID NO:1-11 or SEQ ID NO:14-36. In further embodiments, this disclosure provides the use of the above-described polypeptide or immunogenic compositions comprising the polypeptide in generating an immune response against HBV, or in treating or preventing HBV infection.

[0016] In some embodiments, this disclosure provides a viral vector comprising a cytomegalovirus (CMV) vector containing a polynucleotide encoding a sequence encoding one or more amino acid sequences as listed in SEQ ID NO:1-36. In some embodiments, the polynucleotide encodes one or more amino acid sequences comprising one or more of SEQ ID NO:1-11, SEQ ID NO:14-15, and SEQ ID NO:24-26. In some embodiments, the sequence encoded by the polynucleotide is sequenced to improve expression. In some embodiments, this disclosure provides an immunogenic composition comprising a CMV vector encoding a polynucleotide containing a sequence encoding one or more amino acid sequences as listed in SEQ ID NO:1-36. In further embodiments, this disclosure provides the use of the above-described vector or an immunogenic composition comprising the vector in generating an immune response against HBV, or in treating or preventing HBV infection.

[0017] This document also provides immunogenic compositions or vaccines comprising one or more HBV epitope cosequence antigens. In some embodiments, the antigen of the vaccine is provided as a polypeptide. In some embodiments, the antigen is encoded by a polynucleotide. In some embodiments, the viral vector comprises a polynucleotide. In some embodiments, the antigen is encoded by two or more polynucleotides that can be expressed by the same or different promoters. In some embodiments, the antigen is encoded by different viral vectors. In some embodiments, the HBV vaccine comprises two or more HBV epitope cosequence antigens. In some embodiments, the HBV vaccine comprises a cytomegalovirus (CMV) vector and a polynucleotide encoding one or more HBV epitope cosequence antigens.

[0018] In some implementations, the vaccine is a preventative vaccine. In other implementations, the vaccine is a therapeutic vaccine.

[0019] In some implementations, the immunogenic composition or vaccine further comprises a pharmaceutically acceptable carrier or excipient.

[0020] In some aspects, this disclosure provides a composition, such as an immunogenic composition, comprising two or more of the above-described polypeptides, polynucleotides, vectors, or vaccines.

[0021] This article also provides methods for treating HBV in subjects, the methods comprising administering an effective amount of the aforementioned peptide, vector, vaccine, or composition to subjects in need of such treatment. This article further provides methods for protecting subjects from HBV infection, the methods comprising administering an effective amount of the aforementioned peptide, vector, vaccine, or composition to subjects in need of such treatment. This article further provides methods for generating an immune response against HBV, the methods comprising administering an effective amount of the aforementioned peptide, vector, vaccine, or composition to subjects in need of such treatment.

[0022] This article also provides some implementation schemes in which the above-mentioned peptides, vectors, vaccines or compositions are used to treat HBV, protect subjects from HBV infection, or induce an immune response to HBV.

[0023] This disclosure also provides the use of the above-described polypeptides, vectors, vaccines, or compositions in the preparation of medicaments for treating HBV infection. This disclosure further provides the use of the above-described polypeptides, vectors, vaccines, or compositions in the preparation of medicaments for protecting subjects from HBV infection. This disclosure further provides the use of the above-described polypeptides, vectors, vaccines, or compositions in the preparation of medicaments for generating or inducing an immune response against HBV.

[0024] This article provides HBV polypeptides comprising one or more epitope cosequencing antigens having amino acid sequences derived from HBVC, S, and P, including full-length sequences, regions thereof, or any combination thereof.

[0025] This document further provides immunogenic compositions or vaccines comprising one or more pharmaceutically acceptable vectors and one or more epitope cosequence antigens. This document also provides immunogenic compositions or vaccines comprising a vector capable of expressing one or more epitope cosequence antigens. In some embodiments, the HBV epitope cosequence antigen comprises two or more epitope cosequences. In some embodiments, the HBV vaccine is a prophylactic vaccine. In some embodiments, the HBV vaccine is a therapeutic vaccine.

[0026] This disclosure further provides a method for preventing or treating HBV infection in a subject, the method comprising administering an effective amount of the above-described immunogenic composition or vaccine to a subject in need of such treatment. It further provides a method for designing and manufacturing an immunogenic composition or vaccine for a subject, the method comprising sequencing HBV virus in the subject, selecting a vaccine antigen designed to cover the diversity within the virus present in the subject, and inserting the vaccine antigen into a vector. This document also provides a method for treating HBV infection in a subject, the method comprising administering an effective amount of the disclosed immunogenic composition or vaccine to a subject in need of such treatment.

[0027] This article also provides a method for inducing an effector memory T cell response, the method comprising: (a) designing one or more epitope co-sequence antigens; (b) producing an immunogenic composition or vaccine comprising a CMV backbone and a polynucleotide encoding one or more epitope co-sequence antigens; and (c) administering the vaccine to a subject in need of such a response.

[0028] In some embodiments, the epitope cosequence antigen of the method provided herein comprises one or more amino acid sequences selected from the group consisting of SEQ ID NO:1-11 and SEQ ID NO:14-36. Attached Figure Description

[0029] Figure 1 .HBV genome organization. Unlike other small chronic viruses (such as HIV or HCV) that exhibit high mutation rates leading to quasi-species, HBV's small (3.2kb) DNA genome is more limited in its mutational capacity due to overlapping open reading frames (ORFs).

[0030] Figure 2A-2B The global distribution of 3041 HBV sequences is available from HBVdb. HBV genotypes are distributed worldwide. Only sequences from countries with known sampling are shown on the map.

[0031] Figures 3A-3B The distribution of HBV sequences in Asia and China can be obtained from HBVdb. In Asia, especially in China, approximately 75% of the genotypes are B and C.

[0032] Figures 4A-4B . Figure 4A : Coverage of the entire HBV proteome using one, two, and three epitope cosequences. Figure 4A The diagram shows that single epitope cosequencing provides varying and limited coverage of the HBV genome (top). Using two epitope cosequencing sequences (middle) or three epitope cosequencing sequences (bottom) improves coverage. Figure 4BThree epitope cosequences provide better coverage than one or two epitope cosequences, as shown by the number of exact decimal matches.

[0033] Figures 5A-5C Overall proteome coverage using one, two, or three epitope cosequences developed from the global HBV population. The use of two or three epitope cosequences developed from the global HBV population significantly improved coverage of all major genotypes compared to using epitope cosequences developed from the B and C HBV populations, while providing comparable coverage for B and C.

[0034] Figure 6 The entire HBV proteome was covered using co-sequences of two epitopes relative to four conserved regions of the HBV genome. Hypervariable regions, including 29 N-terminal amino acids and the N-terminus of the P protein, were shown using windows. Hypervariable regions were removed from further analysis.

[0035] Figures 7A-7B When a ( Figure 7A ) or two ( Figure 7B Epitope cosequences are used in Chinese samples, non-Chinese samples, and global sample cohorts to show the coverage of potential T-cell epitopes (PTEs). The X-axis represents the percentage of coverage. The gray histogram represents the coverage of potential T-cell epitopes from 1000 randomly selected natural strains.

[0036] Figure 8 Comparison of coverage rates for various epitope cosequences. The 2_ChinaGlobal_Epi (2_CHGL_epi) sequence provides 0.809 (80.9%) exact full sequence coverage for the global sample. The 2_CHGL_epi sequence provides 0.879 (87.9%) exact full sequence coverage for the Chinese sample and 0.772 (77.2%) exact full sequence coverage for the non-Chinese sample.

[0037] Figures 9A-9B Chinese Sample ( Figure 9A (left) Non-Chinese samples ( Figure 9A (China) and global sample cohorts ( Figure 9A Comparison of conservative region coverage of 1_D.vac, 1_C.ref, 1_CH_epi, 1_GL_epi, 2_CH_epi, 2_CHGL_epi and 3_GL_epi in the right region. Figure 9B The conserved region coverage of 1_D.vac, 1_GL_epi and 2_CHGL_epi in all major genotypes was depicted.

[0038] Figures 10A-10BCompared with the wild-type genotype D sequence and the 1_GL_epi sequence, 1089 Chinese HBV sequences using the 2_CHGL_epi sequence ( Figure 10A ) and all 3041 global HBV sequences ( Figure 10B Conserved region coverage. Genotype D sequence is used in vaccines by Transgene and GlobeImmune.

[0039] Figure 11A-11D . Figure 11A and Figure 11B This demonstrates the co-sequence antigen of the HBV genotype D epitope base, developed in the United States and Europe for effective epitope coverage; the antigen construct is shown in Figure 11A Start in and in Figure 11B Continued. HBV genotype D epitope cosequence antigens derived from the core (C), PreS1 and PreS2, surface antigen (S), and polymerase (P) sequences are shown in the specified order. Variants of epitope cosequence antigens derived from antigens with missing sequences in the transmembrane (TM) domain or with antigens having rearranged relative to the protein sequence in HBV are developed to minimize aggregation and maximize expression. Figure 11C Constructs of the same antigen are shown, each including a hemagglutinin (HA) epitope tag inserted at the C-terminus. Figure 11D Viral expression of the antigen, as measured by SDS-polyacrylamide gel electrophoresis (SDS-page), was shown, followed by protein immunoblotting developed using antibodies targeting HA epitope tags engineered into each construct.

[0040] Figure 12A-12B . Figure 12A The HBV genotype D epitope cosequence antigen, derived from PreS1 and PreS2, surface antigen (S), polymerase (P), and / or core (C) sequences, is shown. As illustrated, four of the antigens are deleted in the sequence derived from the P protein. A V5 or hemagglutinin (HA) epitope tag was engineered into the construct; the V5 tag was inserted at the N′-terminus, and the HA tag was inserted at the C′-terminus. Figure 12B The viral expression of the antigen, as measured by SDS-page, was shown, followed by a protein immunoblot developed using antibodies targeting V5 or HA epitope tags engineered into each construct. Detailed Implementation

[0041] I. definition

[0042] Various terms related to the described aspects are used throughout the specification and claims. Unless otherwise stated, such terms are to be given their ordinary meaning in the art. Other specifically defined terms will be interpreted in a manner consistent with the definitions provided herein.

[0043] As used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural indicators unless otherwise expressly specified. Thus, for example, reference to “an element” includes a combination of two or more elements, etc. The use of alternatives (e.g., “or”) should be understood to mean any one, both, or any combination of the alternatives, and may be used synonymously with “and / or.” As used herein, the terms “comprising” and “having” are used as synonyms, and these terms and variations thereof are intended to be interpreted as non-limiting.

[0044] As used herein when referring to measurable values ​​such as quantity, duration, etc., the term "about" is intended to cover variations from the specified value by up to ±20%, as such variations are suitable for performing the disclosed methods. Unless otherwise stated, all figures expressing the quantity of components, characteristics such as molecular weight, reaction conditions, etc., as used in the specification and claims should be understood to be modified by the term "about" in all cases. Therefore, unless stated to the contrary, the numerical parameters set forth in the following specification and appended claims are approximate values ​​that may vary depending on the desired characteristics sought to be obtained. At least not in an attempt to limit the application of the equivalence principle to the scope of the claims, each numerical parameter should be interpreted at least according to the number of significant figures reported and by applying ordinary rounding techniques.

[0045] Although the numerical ranges and parameters described in this disclosure are approximate, the numerical values ​​described in the specific embodiments are reported as precisely as possible. However, any numerical value inherently contains some errors, which are necessarily due to the standard deviations found in their respective test measurements.

[0046] The term "comprising" means the presence of the features, integers, steps, or components mentioned in the claims, but does not exclude the presence or addition of one or more other features, integers, steps, components, or groups thereof. The term "consistent substantially of" limits the scope of the claims to the specified materials or steps, as well as those materials or steps that do not materially affect the essential and novel features of the claimed subject matter.

[0047] The word “substantially” does not exclude “completely”; for example, a composition that is “substantially free” of Y can be completely free of Y. The word “substantially” may be omitted from the definitions provided herein where necessary.

[0048] As used above and throughout this disclosure, the term "effective amount" refers to the amount, in terms of dosage and time period, that is effective in achieving the desired therapeutic outcome for a given condition, symptom, or side effect. It should be understood that the effective amount of a component will vary from patient to patient, not only due to the specific vaccine, component, or composition chosen, the route of administration, and the component's ability to produce the desired outcome in an individual, but also due to factors such as: the condition or severity of the symptom to be alleviated, hormone levels, age, sex, individual weight, patient survival status, and the severity of the pathological condition being treated, any medications the patient is concurrently taking or any special diet subsequently followed, and other factors that a person skilled in the art would recognize. The appropriate dosage is determined at the discretion of the attending physician. Dosing regimens may be adjusted to provide an improved therapeutic response. An effective amount is also the amount in which any toxic or adverse effects of the component are outweighed by the therapeutically beneficial effects.

[0049] The term "application" refers to the direct application of a compound or composition, or the application of a prodrug, derivative, or analogue that will form an equal amount of an active compound or substance in the body.

[0050] The terms “subject,” “individual,” and “patient” are used interchangeably herein and refer to an animal, such as a human, that has received treatment (including prophylactic treatment) with the pharmaceutical compositions disclosed herein. As used herein, the term “subject” refers to both human and non-human animals. The term “non-human animal” includes all vertebrates, such as mammals, including non-human primates (particularly higher primates), sheep, dogs, rodents (e.g., mice or rats), guinea pigs, goats, pigs, cats, rabbits, cattle, horses, and non-mammals such as reptiles, amphibians, chickens, and turkeys.

[0051] The term "epitope-based cosequence" refers to a co-occurring sequence based on an epitope. It is a sequence whose epitope matches as closely as possible to an epitope in a set of reference natural sequences. Typically, in the case of a k-character subsequence of a longer natural or vaccine antigen sequence, the terms "epitope" and "potential epitope" refer to a k-character sequence (usually k is in the range of 8-12). T cells can recognize such peptides in an immune response.

[0052] The term “EpiGraph” refers to a computational strategy developed to create sequences that provide optimal epitope cosequences, or a set of sequences that, when combined, provide optimal coverage of different viral sequence populations. The EpiGraph method was previously described in PCT application WO 2016 / 054654A1 and Theiler, et al., Sci. Rep. 6:33987 (2016), the entire contents of which are incorporated herein by reference. The EpiGraph method produces sets of artificial but complete antigens with maximum coverage of potential T-cell epitopes (PTEs, typically 9-mer peptide sequences) found in different viral populations. EpiGraph represents a next step compared to existing mosaic vaccine design methods. Graph-based EpiGraph methods are computationally much more powerful than mosaic methods using genetic algorithms, thus allowing for improved PTE coverage by considering significantly more combinations. Both EpiGraph and mosaic methods produce protein antigens with greater T-cell epitope diversity coverage than natural strains. Compared to combinations of natural strains or shared immunogens, HBV, HIV, and HCV mosaics elicit cellular immune responses with greater breadth and depth. HBV and HIV mosaics are currently in Phase I clinical trials. In some implementations, the EpiGraph algorithm is used to design “epitope co-sequences” corresponding to conserved regions of HBV.

[0053] The term "epitope cosequence" refers to the amino acid sequence of an artificial antigen designed using the EpiGraph algorithm based on an HBV sequence population. The epitope cosequence, serving as the "core" of the HBV sequence population, is the calculated sequence that provides the maximum average epitope coverage for that population. An "epitope cosequence antigen" is an antigen containing the epitope co-amino acid sequence.

[0054] As used herein, the term “treatment” or “therapy” (and its various forms, including curative or palliative) refers to the treatment of an infected person. As used herein, the term “treatment” includes the relief or reduction of at least one adverse or negative effect or symptom of a condition, disease, or illness. Such a condition, disease, or illness may be HBV infection.

[0055] As used herein, the terms “prevention” or “preventive” refer to preventing or reducing the risk of a subject contracting or contracting a subject contracting, such as HBV or a related virus, or stopping or reducing the risk of transmission of, such as HBV or a related virus.

[0056] "Pharmaceutically acceptable" means compounds, materials, compositions, and / or dosage forms that are suitable for contact with human and animal tissues within the limits of reasonable medical judgment without excessive toxicity, irritation, allergic reactions, or other problems or complications commensurate with a reasonable benefit / risk ratio.

[0057] As used herein, the terms “vaccine,” “immunogenic compound,” and “immunogenic composition” are used interchangeably to refer to a compound or composition that induces an immune response in a subject. Prophylactic vaccines provide a degree of protection against new infections. Therapeutic vaccines help treat existing infections.

[0058] II. antigen

[0059] This document provides polypeptides comprising sequences derived from certain HBV populations. In some embodiments, the polypeptide comprises amino acid sequences as listed in SEQ ID NO:1-11 or SEQ ID NO:14-36. In some embodiments, the polypeptide comprises two or more amino acid sequences as listed in SEQ ID NO:16-36. In some embodiments, the polypeptide comprises amino acid sequences as listed in SEQ ID NO:19 or 29, amino acid sequences as listed in SEQ ID NO:16, 21, 27, 28 or 34, and amino acid sequences as listed in SEQ ID NO:20, 22, 23, 30, 31, 33, 35 or 36. In some embodiments, the polypeptide comprises an amino acid sequence as listed in SEQ ID NO:1. In some embodiments, the polypeptide comprises an amino acid sequence as listed in SEQ ID NO:2. In some embodiments, the polypeptide comprises an amino acid sequence as listed in SEQ ID NO:3. In some embodiments, the polypeptide comprises an amino acid sequence as listed in SEQ ID NO:4. In some embodiments, the polypeptide comprises an amino acid sequence as listed in SEQ ID NO:5. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:6. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:7. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:8. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:9. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:10. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:11. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:14. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:15. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:24. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:25. In some embodiments, the polypeptide comprises the amino acid sequence listed in SEQ ID NO:26. In some embodiments, the polypeptide comprises both the amino acid sequences listed in SEQ ID NO:3 and SEQ ID NO:4. In some embodiments, the polypeptide comprises the amino acid sequences listed in SEQ ID NO:5 and SEQ ID NO:6. In some embodiments, the polypeptide comprises the amino acid sequences listed in SEQ ID NO:7 and SEQ ID NO:8.In some embodiments, the polypeptide comprises an amino acid sequence as listed in SEQ ID NO:9, an amino acid sequence as listed in SEQ ID NO:10, and an amino acid sequence as listed in SEQ ID NO:11. In any of the above embodiments, the polypeptide may further comprise one or more amino acid sequences as listed in SEQ ID NO:17 and 18.

[0060] In some embodiments, this disclosure provides immunogenic compositions or vaccines comprising one or more of the above-described polypeptides. In some embodiments, the immunogenic composition comprises one of the above-described polypeptides. In other embodiments, the immunogenic composition comprises two or more of the above-described polypeptides, which may be provided as a single polypeptide or as two or more polypeptides.

[0061] In some implementations, the peptide is an epitope-based cosequence antigen comprising a sequence derived from wild-type HBV sequences using the EpiGraph method. In the EpiGraph algorithm, the natural sequence is characterized by a large node graph, where each node corresponds to an epitope appearing in the natural sequence. Directed edges connect two nodes when corresponding epitope strings are “identical” (meaning the last k-1 characters of the first string match the first k-1 characters of the second string). If the two strings are identical, they together form a string of length k+1. More generally, a path through the node and edge graph corresponds to a single string containing a k-merged substring corresponding to each node in the graph. Each node is weighted based on how many sequences in the reference set display the substring corresponding to that node. The EpiGraph algorithm uses a dynamic programming scheme to find paths through the complete graph that maximizes the sum of these weights and provides the maximum coverage.

[0062] In some embodiments, the epitope cosequencing antigen is derived from the HBV C protein. In some embodiments, the epitope cosequencing antigen is derived from the HBV Pol(P) or S protein, or another HBV protein. The epitope cosequencing antigen may be derived from a full-length protein sequence, a region of a full-length protein sequence, or any combination thereof. In some embodiments, the epitope cosequencing antigen comprises two or more epitopes derived from the HBV C, P, and S proteins. In some embodiments, the epitope cosequencing antigen further comprises one or more epitopes derived from the PreS1 domain or the PreS2 domain. In some embodiments, the epitope cosequencing antigen is derived from a conserved region of HBV. In some embodiments, the epitope cosequencing antigen is derived from one or more conserved regions of HBV C, S, and P.

[0063] In some implementations, the epitope cosequencing antigen comprises two or more sequences derived from HBV C, P, and S proteins, wherein the sequences are rearranged relative to their order in the HBV genome. Reordering of sequences within the epitope cosequencing antigen can provide more efficient or otherwise improved in vitro antigen expression.

[0064] In some embodiments, the epitope cosequence antigen is derived from an HBV protein having one or more mutations (e.g., amino acid deletions or substitutions). In some embodiments, the mutation inactivates at least one function present in the wild-type protein. For example, PreS1 can be inactivated by removing the myristylation sequence at the N-terminus of the protein, or Pol can be inactivated by deleting or mutating the enzyme's active site. In some embodiments, one or more mutations are present at the active site of HBV ribonuclease H (RNAseH) and result in loss of RNAseH activity. Exemplary mutations are shown in Table 1.

[0065] In some embodiments, the epitope co-sequence antigen is developed using a selected HBV population, resulting in an epitope co-sequence antigen that serves as the core of that population. For example, the HBV population could be HBV genotypes present in a selected geographic location (e.g., Asia, North America, South America, Europe, Africa, or Australia). In some embodiments, the epitope co-sequence antigen is core to the HBV B regional epidemic in Asia or more specifically, China. In other embodiments, the epitope co-sequence antigen is core to the HBV C regional epidemic in Asia or more specifically, China. In some embodiments, the epitope co-sequence antigen is core to the HBV A regional epidemic in Asia or more specifically, China. In some embodiments, the epitope co-sequence antigen is core to the HBV CB regional epidemic in Asia or more specifically, China. In some embodiments, the epitope co-sequence antigen is core to the HBV D regional epidemic in Asia or more specifically, China. In some embodiments, the epitope co-sequence antigen is core to the HBV DC regional epidemic in Asia or more specifically, China. In some embodiments, the epitope co-sequence antigen is core to the HBV DE regional epidemic in Asia or more specifically, China. In some implementations, the epitope cosequence antigen is central to the regional epidemiology of HBV E in Asia, or more specifically, China. In some implementations, the epitope cosequence antigen is central to the regional epidemiology of HBV F in Asia, or more specifically, China.

[0066] In other embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV multigenotypes in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV A in North America. In some embodiments, population epitope co-sequence antigens are central to the regional epidemics of HBV B in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV C in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV CB in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV D in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV DC in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV DE in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV E in North America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV F in North America.

[0067] In other embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV multigenotypes in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV A in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV B in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV C in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV CB in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV D in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV DC in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV DE in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV E in South America. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV F in South America.

[0068] In other embodiments, epitope co-sequence antigens are central to the European HBV multigenotype regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBV A regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBV B regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBV C regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBV CB regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBV D regional epidemic. In some embodiments, population epitope co-sequence antigens are central to the European HBV DC regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBV DE regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBV E regional epidemic. In some embodiments, epitope co-sequence antigens are central to the European HBVF regional epidemic.

[0069] In other embodiments, epitope co-sequence antigens are central to the regional epidemics of multigenotype HBV in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV A in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV B in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV C in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV CB in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV D in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV DC in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV DE in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV E in Africa. In some embodiments, epitope co-sequence antigens are central to the regional epidemics of HBV F in Africa.

[0070] In other embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV multigenotypes in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV A in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV B in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV C in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV CB in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV D in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV DC in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV DE in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV E in Australia. In some embodiments, the epitope co-sequence antigen is central to the regional epidemic of HBV F in Australia.

[0071] In other implementations, the epitope cosequence antigen is the core of the global HBV set in the disclosed vaccine.

[0072] In some embodiments, the HBV epitope cosequencing antigen is 1_CH_epi (SEQ ID NO:1), which was developed using 1044 HBV sequences from China. In some embodiments, the HBV epitope cosequencing antigen is 1_GL_epi (SEQ ID NO:2), which was developed using a global set of 3041 HBV sequences. In some embodiments, the epitope cosequencing antigen is a combination of two or more individual epitope cosequencing antigens. For example, in some embodiments, the epitope cosequencing antigen comprises 2_CH_epi, which contains two epitope cosequencing antigens developed using 1044 HBV sequences from China (SEQ ID NO:3 and SEQ ID NO:4). In some embodiments, the epitope cosequencing antigen comprises 2_CHGL_epi, which comprises a first epitope cosequencing antigen Epi1 (SEQ ID NO:5) developed using 1044 HBV sequences from China, and a second epitope cosequencing antigen Epi2 (SEQ ID NO:6) developed using a global set of 3041 HBV sequences, wherein Epi1 is already fixed in the solution. In other words, Epi2 is a complement to Epi1 and is developed where Epi1 is already present. In some embodiments, the epitope cosequencing antigen comprises a modified form of 2_CHGL_epi, which comprises Epi3 (SEQ ID NO:7) and Epi4 (SEQ ID NO:8). In some embodiments, the epitope cosequencing antigen comprises 3_GL_epi, which comprises three epitope cosequencing antigens (SEQ ID NO:9; SEQ ID NO:10; and SEQ ID NO:11) developed using a global set of 3041 HBV sequences. In some embodiments, the epitope co-sequence antigen comprises a natural D subtype sequence (SEQ ID NO:12), GenBank accession number Y07587. In some embodiments, the epitope co-sequence antigen comprises a natural C subtype reference sequence (SEQ ID NO:13), GenBank accession number GQ358158. Each reference gene of accession numbers Y07587 and GQ358158 is incorporated herein by reference in its entirety. In some embodiments, the epitope co-sequence antigen is a modified genotype D sequence of SEQ ID NO:14. In some embodiments, the epitope co-sequence antigen is a reordered genotype D sequence of SEQ ID NO:15. In some embodiments, the epitope co-sequence antigen comprises one or more sequences listed in SEQ ID NO:16-23 and SEQ ID NO:27-36. In some embodiments, the HBV epitope co-sequence antigen comprises two or more of the sequences listed in SEQ ID NO:1–SEQ ID NO:36.

[0073] This document further provides immunogenic compositions or vaccines comprising one or more of the above-described epitope cosequencing antigens. The epitope cosequencing antigens can be delivered as DNA, RNA, or peptides. In some embodiments, the vaccine comprises a single antigen. In other embodiments, the vaccine comprises two or more antigens, which may be provided as a single peptide or as two or more peptides. In some embodiments, the vaccine comprises an epitope cosequencing antigen that provides effective epitope coverage for selected HBV genotypes (e.g., genotypes present in selected geographic locations). The vaccines provided herein can be prophylactic or therapeutic vaccines. In some embodiments, the vaccine comprises one or more peptides and a pharmaceutically acceptable carrier or excipient.

[0074] In some embodiments, the vaccine comprises two epitope cosequence antigens developed using HBV sequences from China. In some embodiments, the vaccine comprises an epitope cosequence antigen comprising the sequences listed in SEQ ID NO:3 and SEQ ID NO:4, which may be provided as one peptide or as two peptides. In some embodiments, the vaccine comprises an epitope cosequence antigen developed using HBV sequences from China and an epitope cosequence antigen developed using HBV sequences from a global set. In some embodiments, the vaccine comprises an epitope cosequence antigen comprising the sequences listed in SEQ ID NO:5 and SEQ ID NO:6, which may be provided as one peptide or as two peptides. In some embodiments, the vaccine comprises an epitope cosequence antigen comprising the sequences listed in SEQ ID NO:7 and SEQ ID NO:8, which may be provided as one peptide or as two peptides. In some embodiments, the vaccine comprises three epitope cosequence antigens developed using HBV sequences from a global set. In some embodiments, the vaccine includes an epitope cosequence antigen comprising the sequences listed in SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11, which may be provided as one polypeptide, as two polypeptides or as three polypeptides.

[0075] III. Polynucleotides and carriers

[0076] In some embodiments, this disclosure provides a polynucleotide sequence encoding a polypeptide. The polynucleotide may be DNA or RNA and may encode any of the aforementioned antigens. In some embodiments, the polynucleotide encodes a polypeptide comprising an amino acid sequence listed in one or more of SEQ ID NO:1-36. In some embodiments, the polynucleotide sequence is codon-optimized for expression in a specific host.

[0077] In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequences listed in SEQ ID NO:1-11 or SEQ ID NO:14-36. In some embodiments, the polynucleotide encodes a polypeptide comprising two or more amino acid sequences listed in SEQ ID NO:16-36. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequences listed in SEQ ID NO:19 or 29, the amino acid sequences listed in SEQ ID NO:16, 21, 27, 28 or 34, and the amino acid sequences listed in SEQ ID NO:20, 22, 23, 30, 31, 33, 35 or 36. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:1. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:2. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:3. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:4. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:5. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:6. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:7. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:8. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:9. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:10. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:11. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:14. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:15. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:24. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:25. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequence listed in SEQ ID NO:26. In some embodiments, the polynucleotide encodes a polypeptide comprising both the amino acid sequences listed in SEQ ID NO:3 and SEQ ID NO:4.In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequences listed in SEQ ID NO:5 and SEQ ID NO:6. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequences listed in SEQ ID NO:7 and SEQ ID NO:8. In some embodiments, the polynucleotide encodes a polypeptide comprising the amino acid sequences listed in SEQ ID NO:9, SEQ ID NO:10, and SEQ ID NO:11. In any of the above embodiments, the polynucleotide may further encode a polypeptide comprising one or more amino acid sequences listed in SEQ ID NO:17 and 18.

[0078] In some embodiments, this disclosure provides a composition, such as an immunogenic composition, comprising two or more of the aforementioned polynucleotides.

[0079] In some embodiments, this disclosure provides a vaccine comprising one or more of the aforementioned polynucleotides. In some embodiments, the vaccine further comprises a pharmaceutically acceptable carrier or excipient.

[0080] This article further provides vectors containing any of the aforementioned polynucleotides. Vectors that can be used include, but are not limited to, plasmids, bacterial vectors, and viral vectors. Viral vectors include cytomegalovirus vectors. One advantage of CMV vectors for therapeutic vaccine delivery is that they can be used to elicit specific CD8+ T cell responses and induce more effective and durable responses. Animal models have shown that vaccines based on these viral vectors can clear viral infections (Hansen et al., Science 340:1237874 (2013)), thus these approaches hold promise for therapeutic vaccines (see also: Hansen et al., Science 351:714-720 (2016)).

[0081] Other viral vectors may include poxviruses, including Ankara cowpox and canarypox; adenoviruses, including adenovirus type 5 (Ad5); rubella; Sendai virus; rhabdovirus; alpha virus; and adeno-associated virus. Alternatively, vaccine antigens may be delivered as DNA, RNA, or protein components of the vaccine. Epitope co-sequence antigens will be compatible with virtually any vaccine antigen delivery method.

[0082] In some aspects, this disclosure provides vector-based immunogenic compositions or vaccines, wherein the expression vector is used to deliver nucleic acids encoding one or more antigens disclosed herein. For example, the expression vector may be a poxvirus, adenovirus, rubella virus, Sendai virus, rhabdovirus, alpha virus, or adeno-associated virus backbone.

[0083] In some embodiments, CMV vectors are used in the compositions and methods disclosed herein. Several aspects make chronic HBV infection particularly suitable for immunotherapeutic intervention via CMV vectors.

[0084] First, most individuals with mature immune systems readily develop an effective immune response and clear acute infection gradually. Since this immune clearance is primarily mediated by cytokine-producing CD8+ T cells, inducing an active hepatic-resident effector CD8+ T cell (TEM) response appears crucial for HBV clearance. Therefore, inducing sustained effector HBV-specific CD8+ T cells in the livers of CHB patients should be sufficient to control and ultimately eliminate HBV. The frequency of TEM induced by CMV-based vectors was particularly high in the livers of inoculated rhesus monkeys (RM) (Hansen et al., Nature 473:523-527 (2011), Supplement). Figure 1 ).

[0085] Secondly, unlike other small chronic viruses (such as HIV or HCV) that exhibit high mutation rates leading to quasi-species status, HBV's small (3.2kb) DNA genome is more limited in its mutational capacity due to overlapping open reading frames (ORFs). Figure 1 ).

[0086] Unfortunately, T cells induced by conventional vaccination methods (e.g., peptide-based, protein-based DNA, or even heterologous primacy-boost T cell induction protocols involving both DNA and poxvirus vectors) become undetectable in the periphery after several months. This may be too short to completely prevent the survival of residual HBV genomes, which could potentially provide a source for viral rebound. In contrast, only CMV vector-induced TEMs persist at a high frequency in circulation and the liver for many years (Hansen et al., Nature 2011).

[0087] In some embodiments, the vector comprises a human CMV (HCMV) vector or a rhesus monkey CMV (RhCMV) vector, which contains an HCMV or RhCMV backbone and one or more polynucleotides encoding an antigen. The one or more polynucleotides may encode any of the aforementioned antigens. In some embodiments, the polynucleotide encodes two or more of the aforementioned antigens. In some embodiments, the vector comprises two or more polynucleotides, each encoding one or more of the aforementioned antigens.

[0088] This disclosure further provides compositions comprising two or more vectors, each vector comprising one or more polynucleotides encoding one or more of the aforementioned antigens.

[0089] In some embodiments, the CMV vector lacks the UL82 gene encoding the interlaminar protein pp71. In some embodiments, the UL82 gene is replaced by one or more polynucleotides encoding one or more of the aforementioned antigens. In some embodiments, the CMV vector lacks the UL128-UL130 gene region. In some embodiments, the CMV vector lacks the UL146-UL147 gene region. In some embodiments, the CMV vector lacks both the UL128-UL130 and UL146-UL147 gene regions. In some embodiments, the CMV vector has both the complete UL128-UL130 and complete UL146-UL147 gene regions. The UL128-UL130 gene region includes the UL128 gene, the UL130 gene, and any region between the UL128 and UL130 genes. The UL146-UL147 gene region includes the UL146 gene, the UL147 gene, and any region between the UL146 and UL147 genes.

[0090] In some respects, this disclosure provides immunogenic compositions or vaccines comprising the aforementioned vectors. Immune responses induced by CMV vectors are not affected by pre-existing anti-vector immunity, thus enabling the continuous use of the same vector against different antigens (Hansen et al., Nature Medicine 15:293-299 (2009)). Partly, the capacity for over-infection is due to the viral inhibitors of MHC-I-mediated antigen presentation to CD8+ T cells (Hansen et al., Science 328:102-106 (2010)). Since almost the entire human population, including most CHB patients, is chronically infected with HCMV, over-infection is an important feature, enabling the use of CMV vectors regardless of the recipient's CMV status. CMV vectors possess unique immunology. A surprising and unexpected feature of certain modified RhCMV / SIV vectors is the discovery that these vectors elicit CD8+ T cells that do not recognize any epitopes that are recognized by conventional MHC1a-restricted CD8+ T cells in response to SIV infection itself, or in response to any other vector platform expressing SIV antigens. Nevertheless, CMV vector-induced CD8+ T cells recognize three times the number of peptides within a given antigen, as illustrated in PCT / US2016 / 017373, the entire contents of which are incorporated herein by reference (see also Hansen et al., Science 2013, 2016).

[0091] The fundamental reason for this remarkable breadth was determined by analyzing the restriction elements of individual peptides for a given antigen (CMV, SIV, TB) in over 100 animals, confirming the striking characteristics of the CMV vectors: each peptide induced by the Rh 68-1 vector was presented by either an MHC-II normally recognized by CD4+ T cells, or by a non-polymorphic MHC-I molecule, MHC-E, which normally binds to the MHC-I-derived peptide VMAPRTLLL (VL9) and acts as a ligand for the repressive NKG2A NK cell receptor. (Notably, all CD4+ T cells were conventional, i.e., MHC-II restricted.) It is noteworthy that the response of strain 68-1RhCMV-induced CD8+ T cells to an overlapping peptide library covering the entire antigen could be completely blocked by the addition of VL9 (inhibiting MHC-E) or the invariant chain-derived CLIP (inhibiting MHC-II), confirming that all CD8+ T cell epitopes were “unconventional” (Hansen et al., Science 2016). While MHC-II and MHC-E-restricted CD8+ T cells have been occasionally observed in other infectious diseases and cancers, the abundance of such T cells in RhCMV-immunized animals is unprecedented and truly paradigm-breaking. Importantly, RhCMV vector-induced MHC-II, MHC-E, and MHC-Ia-atypical (discussed below)-restricted CD8+ T cells recognize SIV-infected CD4+ T cells, suggesting that unconventional antigen presentation can occur in SIV-infected cells, even if SIV fails to initiate such a response. Since many chronic viruses (including HCV) upregulate HLA-E, potentially as a defense against NK cells, this highly conserved MHC molecule represents a novel target for immunotherapy. (HLA-E expression in HBV-infected hepatocytes is little known compared to HCV, but high levels of circulating HLA-E are found in chronic HBV carriers). Therefore, the ability to induce MHC-E-restricted CD8+ T cells opens up the possibility of targeting HBV via this highly conserved restriction element.

[0092] Specific patterns of gene modifications in CMV vectors are associated with a variety of T-cell responses. Unconventional CD8+ T cells are induced by vectors lacking the UL128-UL130 and UL146-UL147 gene regions (Hansen et al., Science 2013, 2016, OHSU2346). Conversely, CD8+ T-cell responses induced by natural RhCMV infection or by UL128-UL130-repaired versions targeting RhCMV (Rh68-1) are conventional, i.e., MHC-I restricted (Hansen et al., Science 2013). However, even conventional CD8+ T-cell responses induced by intact UL128-UL130 vectors are significantly more extensive than those induced by non-CMV vectors (Hansen et al., Science 2013). Furthermore, this broad, conventional CD8+ T cell response is entirely targeted at subdominant (“atypical”) epitopes because the immune evasion gene US11 prevents the induction of T cells that recognize “typical” (i.e., immunodominant) MHC-I epitopes (Hansen et al., Science 2013). Therefore, the genetically modified CMV vector can elicit four distinct CD8+ T cell populations, each recognizing a set of non-overlapping epitopes and categorized as follows: 1. Atypical, MHC-II restricted; 2. Atypical, MHC-E restricted; 3. Conventional MHC-I restricted, atypical (=subdominant); 4. Conventional, MHC-I restricted, typical (=immunodominant). The presence or absence of the UL128-UL130 and UL146-UL147 gene regions determines the conversion from conventional CD8+ T cells to unconventional CD8+ T cells. In the absence of US11, classical CD8+ T cells are induced in both vectors containing UL128-UL130 and UL146-UL147 and vectors without UL128-UL130 and UL146-UL147.

[0093] In some embodiments, this disclosure provides an immunogenic composition or vaccine comprising any of the above-described vectors and one or more polynucleotides encoding an antigen. The one or more polynucleotides may encode any of the above-described antigens. In some embodiments, the polynucleotide encodes two or more of the above-described antigens. The two or more polypeptides may be expressed by the same or different promoters. In some embodiments, the vector comprises two or more polynucleotides, each encoding one or more of the above-described antigens. In some embodiments, the vaccine comprises one or more of the above-described vectors and a pharmaceutically acceptable carrier or excipient.

[0094] III. Treatment

[0095] In some embodiments, a method of treating a subject with HBV is provided, the method comprising administering to the subject in such need an effective amount of an immunogenic composition or vaccine comprising one or more of the above-described peptides, polynucleotides, vectors, or compositions. In some embodiments, the vaccine comprises one or more antigens or polynucleotides encoding antigens selected to effectively cover HBV genotypic diversity within a geographic region. In some embodiments, the vaccine is provided in the form of one or more peptides. In other embodiments, the vaccine is provided in the form of one or more polynucleotides, which may be provided in a recombinant viral vector. In any of the above embodiments, the vaccine may further comprise one or more pharmaceutically acceptable vectors or excipients.

[0096] In some embodiments, methods are provided for inducing or generating an immune response in a subject. Such methods involve administering to a subject in need an effective amount of an immunogenic composition or vaccine comprising one or more of the peptides, polynucleotides, vectors, or compositions described above. In some embodiments, the vaccine comprises one or more antigens or polynucleotides encoding antigens, said antigens being selected to effectively cover HBV genotypic diversity within a geographic region. In some embodiments, the vaccine is provided in the form of one or more peptides. In other embodiments, the vaccine is provided in the form of one or more polynucleotides, which may be provided in a recombinant viral vector. In any of the above embodiments, the vaccine may further comprise one or more pharmaceutically acceptable vectors or excipients.

[0097] Some embodiments include a method of treating HBV infection in subjects, the method comprising administering to a subject in such need an effective amount of an immunogenic composition or vaccine comprising one or more of the above-described peptides, polynucleotides, vectors, or compositions. In some embodiments, the vaccine comprises one or more antigens or polynucleotides encoding antigens selected to effectively cover HBV genotypic diversity within a geographic region. In some embodiments, the vaccine is provided in the form of one or more peptides. In other embodiments, the vaccine is provided in the form of one or more polynucleotides, which may be provided in a recombinant viral vector. In any of the above embodiments, the vaccine may further comprise one or more pharmaceutically acceptable vectors or excipients.

[0098] This disclosure further provides a method for protecting a subject from HBV infection, the method comprising administering to a subject in such need an effective amount of an immunogenic composition or vaccine comprising one or more of the above-described peptides, polynucleotides, vectors, or compositions. In some embodiments, the vaccine comprises one or more antigens or polynucleotides encoding antigens selected to effectively cover HBV genotypic diversity within a geographic region. In some embodiments, the vaccine is provided in the form of one or more peptides. In other embodiments, the vaccine is provided in the form of one or more polynucleotides, which may be provided in a recombinant viral vector. In any of the above embodiments, the vaccine may further comprise one or more pharmaceutically acceptable vectors or excipients.

[0099] Some embodiments include methods for inducing effector memory T-cell responses, the methods comprising identifying one or more epitope cosequences; generating a vaccine comprising a vector containing one or more polynucleotides encoding one or more epitope cosequences; and administering the vaccine to a subject in need of such a response. Some embodiments include methods for inducing effector memory T-cell responses, the methods comprising identifying one or more epitope cosequences; generating a vaccine comprising one or more antigens having an amino acid sequence of one or more epitope cosequences; and administering the vaccine to a subject in need of such a response. In some embodiments, a method for inducing an effector memory T cell response is provided, wherein one or more epitope base cosequences comprise 1_CH_epi (SEQ ID NO:1); 1_GL_epi (SEQ ID NO:2); 2_CH_epi, which contains the amino acid sequences listed in SEQ ID NO:3 and SEQ ID NO:4; 2_CHGL_epi, which contains the amino acid sequences listed in SEQ ID NO:5 and SEQ ID NO:6; 2_CHGL_epi, which contains the amino acid sequences listed in SEQ ID NO:7 and SEQ ID NO:8; 3_GL_epi, which contains the amino acid sequences listed in SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11; a natural D isotype sequence (SEQ ID NO:12); a natural C isotype reference sequence (SEQ ID NO:13); a modified genotype D sequence of SEQ ID NO:14; and a reordered genotype D sequence of SEQ ID NO:15. In some embodiments, a method for inducing an effector memory T cell response is provided, wherein one or more epitope cosequences comprise one or more sequences listed in SEQ ID NO:16-23 and SEQ ID NO:27-36. In some embodiments, a method for inducing an effector memory T cell response is provided, wherein one or more epitope cosequences comprise two or more of SEQ ID NO:1–SEQ ID NO:36.

[0100] Some embodiments include a method for inducing an effector memory T-cell response, the method comprising generating an immunogenic composition or vaccine comprising one or more of the aforementioned polynucleotides encoding one or more epitope cosequence antigens; and administering the immunogenic composition or vaccine to a subject in need of such response. In some embodiments, a method for inducing an effector memory T cell response is provided, wherein one or more epitope base cosequences comprise 1_CH_epi (SEQ ID NO:1); 1_GL_epi (SEQ ID NO:2); 2_CH_epi, which contains the amino acid sequences listed in SEQ ID NO:3 and SEQ ID NO:4; 2_CHGL_epi, which contains the amino acid sequences listed in SEQ ID NO:5 and SEQ ID NO:6; 2_CHGL_epi, which contains the amino acid sequences listed in SEQ ID NO:7 and SEQ ID NO:8; 3_GL_epi, which contains the amino acid sequences listed in SEQ ID NO:9, SEQ ID NO:10 and SEQ ID NO:11; a natural D isotype sequence (SEQ ID NO:12); a natural C isotype reference sequence (SEQ ID NO:13); a modified genotype D sequence of SEQ ID NO:14; and a reordered genotype D sequence of SEQ ID NO:15. In some embodiments, a method for inducing an effector memory T cell response is provided, wherein one or more epitope cosequences comprise one or more sequences listed in SEQ ID NO:16-23 and SEQ ID NO:27-36. In some embodiments, a method for inducing an effector memory T cell response is provided, wherein one or more epitope cosequences comprise two or more of SEQ ID NO:1–SEQ ID NO:36.

[0101] In any of the above embodiments, the immunogenic composition or vaccine may be a preventive vaccine or a therapeutic vaccine.

[0102] Recent breakthroughs in HBV vaccine research include the concept of effector memory T cell (TEM)-induced vaccines to prevent HBV infection. Unlike central memory T cells (TCMs) induced by conventional vaccine methods, TEMs are persistently maintained in lymphoid tissues and extraly lymphoid effector sites, immediately ready to mediate antiviral effector functions, thus providing sustained immune protection at the gateways to viral entry and sites of viral reactivation. The most suitable delivery system for inducing and sustaining TEMs indefinitely is derived from CMVs. It is hypothesized that CMVs maintain the appropriate, sustained, low-level immune stimulation required for TEM maintenance without inducing T cell exhaustion due to persistent low-level reactivation and / or gene expression in persistently infected cells.

[0103] In some embodiments, the immunogenic composition or vaccine can be used in a therapeutic vaccine setting. For example, a k-means clustering strategy can be used to design vaccines to identify a set of 6-10 antigen sequences that can provide good coverage of epitopes in a human population infected with highly variable pathogens (e.g., HBV). In some embodiments, the virus in an infected subject is sequenced and two or more vaccines (or one or more vaccines comprising two or three peptides or one or more polynucleotides encoding two or three peptides) that provide good coverage of epitopes in the infecting virus are delivered. Thus, effective epitope coverage is provided while epitope mismatches between the vaccine and the infecting strain are minimized.

[0104] Some embodiments provided include immunogenic compositions or vaccines comprising an HCMV backbone vector lacking certain CMV gene regions, and a polynucleotide encoding an epitope co-sequence antigen. In some embodiments, the HCMV backbone lacks the UL128-UL130 gene regions (including the UL128 gene, the UL130 gene, and any region between the UL128 and UL130 genes) and the UL146-UL147 gene regions (including the UL146 gene, the UL147 gene, and any region between the UL146 and UL147 genes). Some embodiments may also include the deletion of the mesenchymal protein pp71 (UL82) gene. (US Patent Application Publication Nos. 2014 / 0141038; 2008 / 0199493; 2013 / 0142823; and International Application Publication No. WO / 2014 / 138209).

[0105] In some embodiments, this disclosure provides any of the above-described polypeptides, polynucleotides, vectors, vaccines, or compositions for treating HBV infection. In some embodiments, this disclosure provides any of the above-described polypeptides, polynucleotides, vectors, vaccines, or compositions for protecting a subject from HBV infection.

[0106] This disclosure further provides the use of any of the above-described polypeptides, polynucleotides, vectors, vaccines, or compositions in the preparation of medicaments for treating HBV infection. In some embodiments, this disclosure further provides the use of any of the above-described polypeptides, polynucleotides, vectors, immunogenic compositions, vaccines, or compositions in the preparation of medicaments for protecting subjects from HBV infection.

[0107] Typical routes of administration of the polypeptides, polynucleotides, vectors, vaccines, or compositions described herein include, but are not limited to, oral, topical, transdermal, inhalation, parenteral, sublingual, oral, rectal, vaginal, and intranasal administration. As used herein, the term "parenteral" includes subcutaneous, intravenous, intramuscular, intrasternal injection, or infusion techniques. In some embodiments, administration includes administration via a route selected from oral, intravenous, parenteral, gastric, intrapleural, intrapulmonary, intrarectal, intradermal, intraperitoneal, intratumoral, subcutaneous, topical, transdermal, intracisional, intrathecal, intranasal, and intramuscular administration.

[0108] Pharmaceutical compositions according to certain embodiments of the invention are formulated such that the active ingredient contained therein is bioavailable when the composition is administered to a patient. The composition to be administered to a subject or patient may be in the form of one or more dose units, wherein, for example, a tablet may be a single dose unit, and a container for a composition in the form of a polypeptide, polynucleotide, carrier, vaccine, or liquid described herein may contain multiple dose units. Practical methods for preparing such dosage forms are known or will be apparent to those skilled in the art; see, for example, Remington: The Science and Practice of Pharmacy, 20th edition (Philadelphia College of Pharmacy and Science, 2000). In any case, the composition to be administered will contain an effective amount of the polypeptide, polynucleotide, carrier, vaccine, or composition of this disclosure for the treatment or prevention of HBV according to the teachings herein.

[0109] The composition may be in solid or liquid form. In some embodiments, the carrier is granular, so the composition is, for example, in tablet or powder form. The carrier may be liquid, wherein the composition is, for example, an oral oil, an injectable liquid, or an aerosol, which may be used for, for example, inhalation administration. When intended for oral administration, the pharmaceutical composition is preferably in solid or liquid form, wherein semi-solid, semi-liquid, suspension, and gel forms are included in forms considered solid or liquid herein.

[0110] As a solid composition for oral administration, the pharmaceutical composition can be formulated into powders, granules, compressed tablets, pills, capsules, chewing gum, rice paper wafers, etc. Such solid compositions will typically contain one or more inert diluents or edible carriers. In addition, one or more of the following may be present: binders, such as carboxymethyl cellulose, ethyl cellulose, microcrystalline cellulose, tragacanth gum, or gelatin; excipients, such as starch, lactose, or dextrin; disintegrants, such as alginate, sodium alginate, Primogel, corn starch, etc.; lubricants, such as magnesium stearate or Sterotex; flow aids, such as colloidal silica; sweeteners, such as sucrose or saccharin; flavoring agents, such as peppermint, methyl salicylate, or orange flavoring; and coloring agents. When the composition is in capsule form (e.g., gelatin capsules), in addition to the materials of the types described above, it may also contain a liquid carrier, such as polyethylene glycol or oil.

[0111] The composition may be in liquid form, such as an elixir, syrup, solution, emulsion, or suspension. As two examples, the liquid may be for oral administration or delivery by injection. When intended for oral administration, the preferred composition, in addition to the compounds of the present invention, contains one or more of a sweetener, preservative, dye / coloring agent, and flavor enhancer. In compositions intended for injection administration, one or more of a surfactant, preservative, wetting agent, dispersant, suspending agent, buffer, stabilizer, and isotonic agent may be included.

[0112] Liquid pharmaceutical compositions, whether solutions, suspensions, or other similar forms, may include one or more of the following adjuvants: sterile diluents, such as water for injection; saline solutions, preferably physiological saline, Ringer's solution, or isotonic sodium chloride; fixed oils, such as synthetic mono- or diglycerides of glycerol, polyethylene glycol, glycerol, propylene glycol, or other solvents that can be used as solvents or suspension media; antimicrobial agents, such as benzyl alcohol or methylparaben; antioxidants, such as ascorbic acid or sodium bisulfite; chelating agents, such as ethylenediaminetetraacetic acid; buffers, such as acetate, citrate, or phosphate; and agents for adjusting tonicity, such as sodium chloride or glucose. Parenteral formulations may be packaged in ampoules, disposable syringes, or multi-dose vials made of glass or plastic. Physiological saline is a preferred adjuvant. Injectable pharmaceutical compositions are preferably sterile. Liquid compositions intended for parenteral or oral administration should contain an amount of peptides, polynucleotides, vectors, vaccines, or compositions disclosed herein to obtain an appropriate dosage.

[0113] The composition may be intended for topical application, in which case the carrier may suitably comprise a solution, emulsion, ointment, or gel matrix. For example, the matrix may comprise one or more of the following: petrolatum, lanolin, polyethylene glycol, beeswax, mineral oil, diluents (such as water and alcohol), and emulsifiers and stabilizers. A thickener may be present in the composition for topical application. If intended for transdermal application, the composition may comprise a transdermal patch or an iontophoresis device. The pharmaceutical composition may be intended for rectal administration, for example, in the form of a suppository, which melts in the rectum and releases the drug. Compositions for rectal administration may contain an oily matrix as a suitable non-irritating excipient. Such matrices include, but are not limited to, lanolin, cocoa butter, and polyethylene glycol.

[0114] The composition may include a variety of materials that alter the physical form of the solid or liquid dosage unit. For example, the composition may include a material that forms a coating shell around the active ingredient. The material forming the coating shell is typically inert and may be selected from, for example, sugars, shellac, and other enteric coating agents. Alternatively, the active ingredient may be encapsulated in gelatin capsules. The composition in solid or liquid form may include an agent that binds to the polypeptide, polynucleotide, carrier, vaccine, or composition disclosed herein and thereby facilitates the delivery of the compound. Suitable agents that can function in this way include monoclonal or polyclonal antibodies, one or more proteins, or liposomes. The composition may consist essentially of dosage units that can be administered as an aerosol. The term aerosol is used to refer to a variety of systems, from colloidal systems to systems consisting of pressurized packaging. Delivery can be made by liquefying or compressing gas or by a suitable pump system that dispenses the active ingredient. Aerosols can be delivered in single-phase, two-phase, or three-phase systems to deliver the active ingredient. Aerosol delivery includes the necessary containers, activators, valves, sub-containers, etc., which together can form a kit. Preferred aerosols can be determined by those skilled in the art without extensive experimentation.

[0115] It should be understood that the compositions disclosed herein also cover carrier molecules for polynucleotides, as described herein (e.g., lipid nanoparticles, nanoscale delivery platforms, etc.).

[0116] Pharmaceutical compositions can be prepared using methods well-known in the pharmaceutical industry. For example, a composition intended for injection can be prepared by combining the composition with sterile distilled water to form a solution, the composition comprising one or more of the peptides, polynucleotides, vectors, vaccines, or compositions described herein, and optionally, salts, buffers, and / or stabilizers. Surfactants can be added to promote the formation of a homogeneous solution or suspension. Surfactants are compounds that interact nonvalently with molecules to promote the dissolution or homogeneous suspension of molecules in an aqueous delivery system.

[0117] Typically, the appropriate dosage and treatment regimen provide the composition in an amount sufficient to provide therapeutic and / or preventative benefits.

[0118] The composition is administered in an effective amount, which will vary depending on a number of factors, including the activity of the specific compound used; the metabolic stability and duration of action of the compound; the subject's age, weight, general health status, sex, and diet; the method and time of administration; the rate of excretion; the combination of drugs; the severity of the specific condition or symptom; and the subject receiving treatment.

[0119] The following examples are provided to describe the embodiments described herein in more detail. They are intended to be illustrative and not limiting of the embodiments.

[0120] All documents, patents and patent applications cited herein are incorporated by reference and may be used in the practice of the methods and compositions disclosed herein.

[0121] Example

[0122] Example 1:

[0123] Confirmation of epitope co-sequence antigens for use in therapeutic vaccines

[0124] The development of epitope cosequence antigens against HBV using the EpiGraph method is described in PCT application WO 2016 / 054654 A1 and Theiler, et al., Sci. Rep. 6:33987 (2016) (which is incorporated herein by reference in its entirety). Tools for using the EpiGraph algorithm are also available at hiv.lanl.gov / content / sequence / EPIGRAPH / epigraph.html.

[0125] While constructing a designer vaccine for each participant is not feasible, sequencing the virus from that participant to attempt to obtain a good match from a small reference set of vaccine options is feasible. A trial population based on the Chinese HBV B genotype was used to identify vaccines that could provide therapeutic efficacy. An Asian-based reference vaccine set and a global vaccine set were designed, along with an updated Asian-based genotype set. Conserved regions of the HBV genome were identified in the C protein (minus the first 29 amino acids above the N-terminus), S protein, and P protein. The highly variable N-terminal region of the P protein was removed from consideration (see [link to relevant documentation]). Figure 6 ).

[0126] Unlike preventative vaccines, where it is unknown which viruses the subjects may encounter, in the case of therapeutic vaccines, the sequences of the infecting viruses can be obtained and matched.

[0127] When selecting antigens for immunogenic compositions or vaccines, two factors should be considered: the antigenic epitopes should match as many epitopes as possible with the subject's infecting virus, and the epitope mismatches between the antigen and the subject's infecting virus should be as few as possible so that the vaccine response targets the relevant epitopes as much as possible.

[0128] Phylogenetic relationships within major HBV genotypes often lack a clear structure. Instead, it's a "starburst" where the outer branches are very long, while the inner branches near the base are very short and poorly defined. Part of this structure may result from intra-subtype recombination. While this is difficult to quantify, recombination certainly occurs relatively frequently, and similar to what is seen with inter-subtype recombination, it can be widespread. Given the tree structure, simply using clustering on the phylogenetic tree to define a reference set for potential vaccines is less effective, as intra-genotype associations have limited significance from an epitope perspective. Instead, relevant measures should consider sequence relationships, and the reference set should be selected based on the potential epitope similarities between the natural strain and the putative vaccine design.

[0129] Use amino acid 9-mers (see...) Figure 4A The EpiGraph sequence is confirmed, but this method can use k-mers of any length as a reference point, where k is the estimated potential epitope length. In past mosaicking work, the optimal solution for 9-mers has been nearly optimal for other close lengths (8, 10, 11, 12), and the EpiGraph method is expected to be similar.

[0130] Epitope coverage of a vaccine includes: a) one epitope co-sequence; b) two epitope co-sequences; c) three epitope co-sequences; d) one natural HBV genotype sequence; and e) in Figure 4A-7B The study compared two natural HBV genotype sequences. Typically, genotype-based vaccines provide excellent coverage for that genotype, but poor coverage for other genotypes. Figures 5A-5C Data from [the study] confirms that, compared to B and C specific vaccines, bivalent and trivalent Global EpiGraph sequences significantly improved coverage across all major genotypes while providing comparable B and C coverage. For example, [the data is presented in the original text]. Figures 5A to 5C As shown, when using three epitope co-sequence antigens, the genotype D coverage is approximately 90%, compared to approximately 50% for B or C specific vaccines or even B+C specific vaccines.

[0131] Example 2:

[0132] Transient expression of epitope cosequence antigens

[0133] Antigens designed using the EpiGraph method to maximize epitope matching resemble natural sequences but no longer encode natural proteins. Although theoretical guidelines for expressing these artificial amino acid sequences were followed when constructing them, the proteins may exhibit unexpected expression profiles or fail to express stable full-length proteins.

[0134] To assess the expression profiles of these sequences in mammalian cells, polynucleotides encoding epitope-based cosequence antigens were synthesized and cloned for transient transfection using the methods described in WO2016 / 054654 (the entire contents of which are incorporated herein by reference). Briefly, DNA encoding the constructs (Genscript, Piscataway, NJ) was synthesized to contain compatible cloning sites for plasmid vectors (pcDNA3.1 and pOri). All inserts were codon-optimized for the host. Each construct was also modified to eliminate residual enzymatic activity of the native sequences, as described by Kulkarni et al. (Vaccine, 2011) (the entire contents of which are incorporated herein by reference). Plasmid vectors were linearized with compatible endonucleases and treated with calf intestinal phosphatase to prevent recirculation of the empty vectors. Vectors and inserts were separated by agarose gel electrophoresis to determine digested fragment size and cleaned using a PCR purification kit (Thermo Scientific) for ligation. Using a rapid ligation kit (Roche, Indianapolis, Indiana), the insert was ligated to a linearized vector at an insert-to-vector ratio of approximately 3:1 at room temperature for 15 minutes, transformed into chemically competent *E. coli* (DH5α), and plated on antibiotic-selective plates. DNA from the resulting colonies was screened by restriction digestion of the insert.

[0135] Clones containing each correctly oriented insert relative to the vector promoter and poly(A) sequence were grown in liquid culture for plasmid DNA purification. For liposome preparation, actively growing subfusion 293T or HELA cells in 12-well tissue culture plates were given 500 μl of fresh medium (DMEM 10% FBS). To generate liposomes containing plasmid DNA, 250 μl of serum-free medium was mixed with 500 ng of plasmid DNA, and 250 μl of serum-free medium was mixed with 2 μl of lipid (Lipofectamine 2000, Invitrogen). After incubating at room temperature for 5 minutes, these solutions were combined, mixed, and incubated for 20 minutes. The DNA-containing liposomes (500 μl) formed during this process were added dropwise to the culture and incubated for 12–16 hours, after which the transfection mixture was replaced with fresh medium. After another day of incubation, the culture was harvested by scraping and centrifugation. The supernatant was removed by aspiration, and the cell pellet was lysed by resuspending in 100 μl of gel-loaded dye containing 5% SDS and 10% 2-mercaptoethanol and centrifuging on a QiaShred column (Qiagen, Valencia, California).

[0136] The expression of epitope co-sequence antigens was confirmed by SDS-PAGE (SDS-page) and protein immunoblotting using antibodies developed against V5 or hemagglutinin epitope tags engineered into each construct. Briefly, NuPAGE 4–12% Bis-Tris gels were prepared and loaded with 20–50 μg of total protein, followed by electrophoresis at 110–120 volts for 90 min. The separated proteins were transferred to a PVDF membrane at 30 volts via semi-dry or wet transfer for 90 min. Non-specific binding was blocked for 60 min with a solution of 10% skim milk powder in Tris-buffered saline containing 0.1% Tween-20 (PBS-T). HA (Sigma) or V5 (Invitrogen) antibodies were diluted in 5% milk solution and incubated with the membrane for 1 h, followed by washing three times with TBS-T, and then adding a 1:10000 diluted horseradish peroxidase-conjugated goat anti-mouse (Invitrogen) secondary antibody for 1 h. The blot was then washed three times in TBS-T, developed with enzyme-linked chemiluminescence [ECL kit (Thermo-Pierce)] and visualized using a digital gel imaging system.

[0137] Example 3:

[0138] Epitope cosequence antigens were engineered into the CMV vector BAC construct and expressed from the reconstructed virus.

[0139] Epitope-based cosequencing antigens are designed to provide good coverage of T-cell epitopes representing viral sequence profiles and the HBV genotypes from which they originate. To utilize these antigens most effectively, polynucleotides encoding the antigens have been engineered into CMV vectors, which have been shown to produce three times the CD8+ T-cell profile of competing platforms. The extensive antigen presentation and lifetime expression profile of the CMV vectors have been demonstrated to protect and cure SIV-infected rhesus macaques. When applied to broad-spectrum prophylactic vaccines, epitope-based cosequencing antigen design algorithms combined with CMV vectors can provide even greater HBV coverage both within and across genotypes.

[0140] Using BAC recombinant engineering with galactokinase and kanamycin selection, polynucleotides encoding epitope cosequences confirmed to be expressed in transient transfection systems were transferred to the CMV backbone. (Warming et al. Nucleic Acids Research. 2005. 33(4): 1-12; and Paredes and Yu Curr Protoc Microbiol. 2012. Feb.; Chapter 14: Unit 14E. 4; both are incorporated herein by reference in their entirety). Furthermore, epitope cosequences derived from HBV proteins or protein domains [such as the core (C) protein, surface antigen (S) protein, PreS1 domain, PreS2 domain, and polymerase (P) protein] can be reordered relative to their order of appearance in wild-type HBV, such as... Figures 11A to 11B As shown, this is to improve the expression.

[0141] In the case of *E. coli* strain SW105 containing parental BAC, BAC recombination facilitates the manipulation of large DNA sequences using temperature- and metabolite-regulated recombinases. Recombination is a sequential two-step process consisting of two steps: Step 1: inserting a galactokinase (galK) sequence carrying the antibiotic resistance gene kanamycin (kan) into the target region; and Step 2: replacing the galK / kan cassette with the target antigen. Step 1, following positive selection in kan, produces recombinants containing the galK / kan insert, while Step 2 produces recombinants containing the target antigen following negative selection in 2-deoxygalactose (DOG). For both steps, the sequence to be inserted is amplified from template DNA containing either the galK / kan cassette or the target antigen using primers with long (50+ bp) homologous arms to the HCMV sequence flanking the insertion site via standard PCR.

[0142] To prepare bacterial cells for galK / kan box insertion (step 1), 5 mL of culture was incubated overnight at 30°C in 2xYT or Terrific Broth with 12.5 μg / mL chloramphenicol, and diluted 1:50 the following morning. The bacteria were then allowed to regrow at 30°C for approximately 2–4 hours (to reach OD). 600 =0.5-0.6), and then heat shock was performed by shaking at 42°C for 15 minutes to induce recombinase. After this induction, the bacteria were pelleted (3000 rpm, 10 min, 4°C) and then washed three times in ice-cold water. E. coli cells were made electrocompetent to receive PCR products and recombinant competent to insert sequences into the target region of BAC. The purified insert (300 ng) was bound to 40 μl of competent E. coli on ice, transferred to a 1 mm cuvette, and electroporated using a Bio-rad Gene Pulser Xcell. After electroporation, the bacteria were diluted by adding 5 mL of culture medium and allowed to recover by shaking at 30°C for 2 hours, then plated on chloramphenicol / kanamycin plates. The plates were incubated at 30°C for two days, and colonies were screened for recombination events by restriction digestion and PCR.

[0143] The galK / kan insert-positive BAC constructs proceeded to the second step, where the galK / kan cassette was replaced by recombination with a PCR fragment containing the target antigen. To prepare bacterial cells to replace the galK / kan cassette in step 2, bacteria were grown and electroporated as described above. For electroporation recovery, the bacteria were diluted by adding 5 mL of medium and allowed to recover by shaking at 30°C for at least 4 to 4.5 hours. The cell pellet was then washed three times with 1xM9 medium and plated on M63 basic medium plates supplemented with glycerol, leucine, biotin, DOG, and chloramphenicol. Colonies were then screened by PCR to confirm replacement of the galK / kan cassette with the target antigen. Positive clones were re-stripened on DOG plates and BAC DNA was isolated for further characterization by restriction digestion and sequencing.

[0144] Viral Reconstitution: To regenerate the virus, BAC DNA was transferred to mammalian host cells that allow viral growth. Briefly, BAC DNA (Macherey-Nagel) was purified using an endotoxin-free plasmid DNA kit and transfected (16–24 μg BAC DNA / T150) into confluence flasks of primary human fibroblasts grown at 37°C in DMEM with 1x glutamax and 9% FBS. Transfection was performed using Lipofectamine 3000 according to the manufacturer's protocol (ThermoFisher). The medium was changed the next day, and plaque formation was monitored daily. Once complete cytopathic effect (CPE) was achieved, the viral supernatant was harvested, clarified by centrifugation at 2500 x g for 5 min at room temperature, and stored at -80°C. Remaining attached cells were harvested in DPBS using a cell scraper, precipitated by centrifugation (2,500 rpm, 5 min), and stored at -80°C.

[0145] Viral epitope co-sequence antigen expression: The expression of epitope co-sequence antigens was detected by SDS-PAGE (SDS-page), followed by protein immunoblotting using HBV-specific antibodies or antibodies targeting the V5 or hemagglutinin (HA) epitope tags engineered into each construct. Briefly, cell pellets were lysed by resuspending in radioimmunoprecipitation assay (RIPA) buffer, and proteins were quantified using a standard dioctanine assay (BCA). NuPAGE 4–12% Bis-Tris gels were prepared and loaded with 20–50 μg of total protein and electrophoresed at 110–130 volts for 90 min. The separated proteins were transferred to PVDF membranes by semi-dry or wet transfer at 30 volts for 90 min or overnight transfer at 15 V. Non-specific binding was blocked for 60 min with a solution of 5% skim milk powder in Tris-buffered saline with 0.1% Tween-20 (PBS-T). The primary antibody was diluted in 5% milk solution and incubated with the membrane for 1 hour, followed by washing three times with TBS-T. Then, horseradish peroxidase-conjugated goat anti-mouse secondary antibody (1:10,000 dilution) was added and incubated for 1 hour. Subsequently, the blot was washed three times with TBS-T, developed with enzyme-linked chemiluminescence (ECL kit (Thermo-Pierce)) and visualized using a digital gel imaging system.

[0146] Example 4:

[0147] Population epitope co-sequence vaccine

[0148] The EpiGraph method was originally used to generate a set of multinucleotide vaccines containing a co-sequence antigen encoding an epitope base using a CMV vector; however, other vaccine delivery systems can also be used.

[0149] A total of 58 strategies with different epitope co-sequence antigens and antigen combinations were tested to determine the most comprehensive vaccine treatment. HBV samples from dbHBV were divided into three groups: Chinese samples, non-Chinese samples, and global samples. The global samples included combinations of Chinese and non-Chinese samples. Vaccines targeting natural D and natural C subtypes were used as controls. Vaccines with one, two, or three epitope co-sequence antigens were tested for each group.

[0150] The controls include vaccines containing polynucleotides encoding the following: a natural D subtype sequence (SEQ ID NO:12), GenBank accession number Y07587; and a natural C subtype reference sequence (SEQ ID NO:13), GenBank accession number GQ358158. The vaccines containing polynucleotides encoding subtypes D and C were then compared with vaccines containing polynucleotides encoding the following: (a) 1_CH_epi (SEQ ID NO:1), developed using 1044 HBV sequences from China; (b) 1_GL_epi (SEQ ID NO:2), developed using a global set of 3041 HBV sequences; (c) 2_CH_epi, which contains two epitope cosequencing antigens (SEQ ID NO:3 and SEQ ID NO:4) developed using 1044 HBV sequences from China; (d) 2_CHGL_epi, which contains two epitope cosequencing antigens – the first epitope cosequencing antigen Epi1 (SEQ ID NO:5), developed using 1044 HBV sequences from China, and the second epitope cosequencing antigen Epi2 (SEQ ID NO:6), developed using a global set of 3041 HBV sequences, wherein Epi1 was already fixed in the solution; (e) Epi7 and Epi8 (SEQ ID NO:6 and SEQ ID NO:7), respectively. (f) SEQ ID NO:7 and SEQ ID NO:8), which are variants of Epi1 and Epi2 modified to induce better expression; and (f) 3_GL_epi, which contains three epitope co-sequence antigens (SEQ ID NO:9; SEQ ID NO:10; and SEQ ID NO:11), developed using a global set of 3041 HBV sequences.

[0151] like Figure 8 As shown, the dual epitope co-sequence antigen vaccine, which contains a first sequence encoding a multinucleotide developed using samples from China and a second sequence developed using a global set, provides coverage of ~88% Chinese sequences, ~77% non-Chinese sequences, and ~80% global sequences.

[0152] Example 5:

[0153] Epitope coverage

[0154] The HBV epitope coverage of vaccines containing certain epitope co-sequence antigen sequences was calculated and analyzed. Vaccines used for preliminary CMV testing included: 1) a single population epitope co-sequence antigen, which is central to the Chinese genotype epidemic; 2) a single population epitope co-sequence antigen providing coverage of all global HBV samples; 3) two population epitope co-sequence antigens providing coverage of HBV samples from China; 4) two population epitope co-sequence antigens providing coverage of all global HBV samples; 5) three population epitope co-sequence antigens providing coverage of all global HBV samples; and 6) two population epitope co-sequence antigens: a first population epitope co-sequence antigen providing coverage of HBV samples from China, and a second population epitope co-sequence antigen providing coverage of HBV samples from a global set.

[0155] The controls include vaccines containing polynucleotides encoding the following: a natural D subtype sequence (SEQ ID NO:12), GenBank accession number Y07587; and a natural C subtype reference sequence (SEQ ID NO:13), GenBank accession number GQ358158. We then compared vaccines containing polynucleotides encoding subtypes D and C with vaccines containing polynucleotides encoding the following: (a) 1_CH_epi (SEQ ID NO:1), developed using 1044 HBV sequences from China; (b) 1_GL_epi (SEQ ID NO:2), developed using a global set of 3041 HBV sequences; (c) 2_CH_epi, containing two epitope cosequencing antigens (SEQ ID NO:3 and SEQ ID NO:4) developed using 1044 HBV sequences from China; (d) 2_CHGL_epi, containing two epitope cosequencing antigens – the first epitope cosequencing antigen Epi1 (SEQ ID NO:5), developed using 1044 HBV sequences from China, and the second epitope cosequencing antigen Epi2 (SEQ ID NO:6), developed using a global set of 3041 HBV sequences, wherein Epi1 is already fixed in the solution; (e) Epi7 and Epi8 (SEQ ID NO:6 and SEQ ID NO:7 respectively). (f) SEQ ID NO:7 and SEQ ID NO:8), which are modified variants of Epi1 and Epi2 to induce better expression; and (f) 3_GL_epi, which contains three epitope co-sequence antigens (SEQ ID NO:9; SEQ ID NO:10; and SEQ ID NO:11), developed using a global set of 3041 HBV sequences.

[0156] like Figures 9A to 10BAs observed, the vaccine containing 2_CHGL_epi provides over 80% accurate coverage of sequences from China, over 75% accurate coverage of sequences not from China, and 80% accurate global coverage. These data confirm extremely high coverage of all HBV sequences compared to the control vaccine.

[0157] Example 6:

[0158] Second-generation HBV epitope co-sequence antigen

[0159] For most HBV genotypes, the HBV N-terminal assembly domain (NTD) of the core (C) is involved in core particle assembly, while the C-terminal domain is involved in the packaging of the pregenomic / reverse transcriptase complex. The surface (S) protein is the product of a single open reading frame and is distinguished by three domains: PreS1, PreS2, and S. The polymerase (P) protein exhibits both DNA-dependent DNA polymerase and RNA-dependent DNA polymerase (reverse transcriptase) activities. The P protein replicates the HBV genome from the encapsulated pregenomic RNA template. The P protein consists of four domains: (1) the terminal protein (TP) domain, which participates in the protein initiation mechanism through conserved tyrosine residues; (2) the non-conserved spacer domain; (3) the reverse transcriptase domain [RNA-dependent DNA polymerase (RT) and DNA-dependent DNA polymerase (active site: YMDD conserved motif)]; and (4) the RNase H domain (ribonuclease H activity).

[0160] Epitope-based cosequencing antigens are designed to provide good coverage of T-cell epitopes, which represent the viral sequence profiles from which they originate. HBV genotype D infection is one of the most common HBV infections in the United States and Europe. Therapeutic vaccines containing genotype D epitope-based cosequencing (EpiD) antigens or nucleotides encoding EpiD antigens can benefit patients who have been pre-screened for a specific genotype. This article provides an example of an HBV genotype D epitope-based cosequencing antigen (SEQ ID NO:14) that provides good coverage of HBV genotypes in the United States and Europe. In one variant, such as Figure 11A and 11BAs shown, the HBV genotype D epitope cosequence antigen, derived from the core (C) protein, PreS1 and PreS2 domains, surface antigen (S) protein, and polymerase (P) protein, was initially used in the order of their appearance in the HBV genome: CSP (“CSP”) (SEQ ID NO:14) and reordered to SPC (“SPC”) (SEQ ID NO:15). Additional antigen variants were generated by applying the EpiGraph method to HBV protein sequences in which transmembrane domains 1-2 or 1-4 of the S protein were missing (“CSPΔTM1-2,” “CSPΔTM1-4,” “SPCΔTM1-2,” or “SPCΔTM1-4”) to reduce aggregation and improve expression. Therefore, the HBV epitope cosequence antigen can also be derived from HBV protein sequences containing one or more of the aforementioned variants.

[0161] Furthermore, epitope cosequence antigens derived from P protein variants with mutations and / or deletions in the active site of the polymerase domain are generated to reduce potential toxicity and improve safety. Examples of such epitope cosequence antigens are provided, for example, in SEQ ID NO:7, 8, and 15. Mutations present in the sequences used to develop these epitope cosequence antigens are provided in Table 1 below. Other P protein variants with different mutations or deletions may also be used.

[0162] Table 1: Mutant modifications of polymerase in HBV genotype D

[0163] Location amino acids Modification 538-541 YMDD Missing 689 D Replace with A 718 E Replace with A 737 D Replace with V 777 D Replace with A

[0164] The positions in the left column of this table refer to the number of amino acids in the full-length polymerase of HBV genotype D. The YMDD sequence is the active site of the reverse transcriptase. (See Radziwill et al., J Virol. 1990 Feb; 64(2):613-20, which is incorporated herein by reference in its entirety.) The other four amino acids (D, E, D, D) are the active sites of RNase H. (See Tavis et al., PLoSPathog. 2013 Jan; 9(1):e1003125, which is incorporated herein by reference in its entirety.)

[0165] Expression of epitope cosequence antigens derived from these variant sequences was tested using SDS-page and Western blotting as described above. Figures 11A to 11D Removing the transmembrane (TM) domain does not affect expression. Figure 11D ).

[0166] No significant aggregation of any EpiD antigen variants was observed. The protein sequence of antigen fusions is associated with efficient expression. SPC-reordered antigen fusion proteins showed significantly increased expression.

[0167] HBV epitope cosequence antigens can be derived from one or more of the following HBV proteins or protein domains: core (C) protein, surface (S) protein, PreS1 protein, PreS2 protein, transmembrane domains 1-4 (TM1-4) of the S protein, determinant clusters, and polymerase (P) protein. Epitope cosequence antigens can be derived from various HBV proteins and protein domains with deletions or mutations, and / or epitope cosequence antigens can be rearranged relative to their order in HBV to improve expression or activity. For illustrative purposes, the epitope cosequence derived from the core protein of HBV subtype D is provided in SEQ ID NO:16, the epitope cosequence derived from the PreS1 protein of HBV subtype D is provided in SEQ ID NO:17, the epitope cosequence derived from the PreS2 protein of HBV subtype D is provided in SEQ ID NO:18, the epitope cosequence derived from the surface protein (S) of HBV subtype D is provided in SEQ ID NO:19, the epitope cosequence derived from the polymerase (P) protein of HBV subtype D is provided in SEQ ID NO:20 or 22, and the epitope cosequence derived from the polymerase (P) protein of HBV subtype D containing the mutations and deletions shown in Table 1 is provided in SEQ ID NO:23. An example of an epitope cosequence derived from the surface protein (S), polymerase protein (P), and core protein (C) of HBV subtype C is provided in SEQ ID NO:24, wherein the order of the antigen sequences is “SPC”. Examples of epitope cosequences derived from P proteins having larger deletion regions than those shown in Table 1 include epitope cosequences derived from HBV subtype C surface protein (S), polymerase protein (P) with deletions of amino acids 612-838, and core protein (C), provided in SEQ ID NO:25; and epitope cosequences derived from HBV subtype D surface protein (S), polymerase protein (P) with deletions of amino acids 601-827, and core protein (C), provided in SEQ ID NO:26. Both SEQ ID NO:25 and 26 are reordered to provide the antigen sequence in the order of “SPC”.

[0168] Other examples of epitope cosequences derived from P proteins with missing regions are shown in Figure 12A The deletions include the absence of amino acids 1-200, 201-400, or 401-600 in the P protein. Figure 12A The expression data of the constructs are shown in Figure 12B middle.

[0169] Further examples of epitope cosequences derived from HBV core proteins are provided in SEQ ID NO:27, 28, and 34. Other examples of epitope cosequences derived from HBV surface proteins are provided in SEQ ID NO:29. Further examples of epitope cosequences derived from HBV polymerase proteins are provided in SEQ ID NO:30-32 and SEQ ID NO:35-36.

[0170] The sequences of HBV proteins and protein domains for any HBV subtype can be determined by sequence alignment with the sequences disclosed herein. For example, the complete genome sequence of the native D subtype of HBV is provided in GenBank accession number Y07587 (incorporated herein by reference in its entirety). The P protein is encoded by nucleotides 1-1625 and 2309-3182 of GenBank reference gene Y07587. The PreS protein is encoded by nucleotides 1-837 and 2850-3182 of GenBank reference gene Y07587. The S domain is encoded by nucleotides 157-837 of GenBank reference gene Y07587. The PreC / C protein is encoded by nucleotides 1816-2454 of GenBank reference gene Y07587. The C protein is encoded by nucleotides 1903-2451 of GenBank reference gene Y07587. In addition, the complete genome sequence of the native C subtype of HBV is provided in GenBank accession number GQ358158 (incorporated herein by reference in its entirety). The P protein is encoded by nucleotides 1-1623 and 2307-3215 of the GenBank reference gene GQ358158. The S protein is encoded by nucleotides 1-835 and 2848-3215 of the GenBank reference gene GQ358158. The PreC / C protein is encoded by nucleotides 1814-2452 of the GenBank reference gene GQ358158. The C protein is encoded by nucleotides 1901-2452 of the GenBank reference gene GQ358158.

[0171] Example 7:

[0172] Vaccine testing

[0173] Vaccines containing computationally designed epitope co-sequence antigens were tested in rhesus monkeys (RM). CMV-based T-cell responses were expected to be broader, thus covering a significantly higher percentage of sequences than previously reported against other vectors. Therefore, even with relatively small animal numbers, sufficient epitope responses should be available to assess the impact of sequence variations on the cross-reactivity potential of the responses. The number and magnitude of all responses to the vaccine were determined using groups of peptides matched to the vaccine. Once the target peptides were identified by using only those that were positive in each animal, the impact of natural variants on each vaccine-responding peptide could be determined. The natural variants tested were based on those found in the reference plot. Nonparametric and computational resampling statistics were used as the primary tools to assess the impact of epitope variations on the reduction of magnitude or elimination of identification. However, these analyses were supplemented by using generalized linear models, if necessary, to explore the impact of more complex interactions on the cross-reactivity of T-cell responses.

[0174] The vaccine suite used for initial CMV testing includes vectors encoding the following: 1) a single population epitope co-sequence antigen, which is the core of the Chinese genotype epidemic; 2) a single population epitope co-sequence antigen providing coverage of all global HBV samples; 3) two population epitope co-sequence antigens providing coverage of HBV samples from China; 4) two population epitope co-sequence antigens providing coverage of all global HBV samples; 5) three population epitope co-sequence antigens providing coverage of all global HBV samples; and 6) two population epitope co-sequence antigens: a first population epitope co-sequence antigen providing coverage of HBV samples from China, and a second population epitope co-sequence antigen providing coverage of HBV samples from the global set; 7) an epitope co-sequence antigen generated using a truncated Pol sequence providing coverage of the HBV subtype epitope (SEQ ID NO:25); and 8) an epitope co-sequence antigen generated using a truncated Pol sequence providing coverage of the HBV subtype epitope (SEQ ID NO:25). Coverage of NO:26). The controls include vaccines containing polynucleotides encoding the following: the natural D subtype sequence (SEQ ID NO:12), GenBank accession number Y07587; and the natural C subtype reference sequence (SEQ ID NO:13), GenBank accession number GQ358158.

[0175] A maximum of 10 rhesus monkeys (RM) in a cohort of 5 each were vaccinated as follows: 10 6PFU HCMV vectors: Up to 8 cohorts, each receiving one of vaccines 1-8 listed above; a single cohort receives a vaccine containing a polynucleotide encoding the natural D subtype; and a single cohort receives a vaccine containing a polynucleotide encoding the natural C subtype. Cohort 3 receives an epitope cosequence plus a different custom vaccine vector, and cohort 4 receives an epitope cosequence plus two custom vaccine vectors.

[0176] Rhesus monkeys (RM) were subcutaneously inoculated at day 0 and week 12 and followed longitudinally for one year. Since vaccination with the HCMV vector is not affected by pre-existing anti-RhCMV immunity, naturally infected RhCMV animals were used for these experiments. Flow cytometry intracellular cytokine analysis (ICS) was used to determine the response of CD4+ and CD8+ T cells to a single, continuous 15-meric peptide containing the vaccine sequence from the vaccine insert administered to each animal (this would contain the total vaccine-induced response). It was then determined whether these epitope-specific T cells recognized epitope variants in both target and non-target strains. For peptides showing a response to strain-specific epitopes, the magnitude, functional affinity, and functional signatures (IFN-γ, TNF-α, IL-2, and MIP-1β production and CD107 exogenization) of these responses to “parental” (vaccine insert) peptide variants were compared to determine the extent of functional cross-reactivity. In selected cases, truncation analysis was used to identify core epitopes for similar comparative analyses. To determine the percentage of MHC-II-restricted CD8+ T cells present, "blocking" mAbs specific to MHC-I and MHC-II, as well as the invariant chain-derived MHC-II-specific binding peptide CLIP, was used to suppress influenza-specific CD8+ T cell responses in PBMCs.

[0177] Although specific implementation schemes have been described and illustrated, it will be readily understood that the various implementation schemes described above can be combined to provide further implementation schemes, and various changes can be made therein without departing from the spirit and scope of the invention.

[0178] Unless otherwise expressly stated, all U.S. patents, U.S. patent application publications, U.S. patent applications, foreign patents, foreign patent applications, and non-patent publications mentioned in the specification and / or listed in the application data sheet, including U.S. Provisional Patent Application No. 62 / 893,546, filed August 29, 2019, and U.S. Provisional Patent Application No. 62 / 941,125, filed November 27, 2019, are incorporated herein by reference in their entirety. If necessary, aspects of the embodiments may be modified to incorporate the concepts of various patents, applications, and publications to provide further embodiments.

[0179] Based on the detailed description above, these and other changes can be made to the embodiments. Generally, the terminology used in the following claims should not be construed as limiting the claims to the specific embodiments disclosed in the specification and claims, but should be interpreted to include all possible embodiments and the full scope of equivalents enjoyed by such claims. Therefore, the claims are not limited by this disclosure. sequence list <110> Weir Biotechnology Co., Ltd. Tsui De National Security LLC <120> Hepatitis B virus vaccine <130> 930185.414WO <140> PCT <141> 2020-08-28 <150> US 62 / 941,125 <151> 2019-11-27 <150> US 62 / 893,546 <151> 2019-08-29 <160> 36 <170> FastSEQ for Windows version 4.0 <210> 1 <211> 1057 <212> PRT <213> Artificial sequence <220> <223> Synthetic sequence 1_CH_epi <400> 1 Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys Ile 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg Gln 180 185 190 Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe Pro 195 200 205 Asp His Gln Leu Asp Pro Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp 210 215 220 Trp Asp Phe Asn Pro Asn Lys Asp His Trp Pro Glu Ala Asn Gln Val 225 230 235 240 Gly Ala Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Gly Leu 245 250 255 Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Thr Thr Val Pro Ala 260 265 270 Ala Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr 275 280 285 Pro Ile Ser Pro Pro Leu Arg Asp Ser His Pro Gln Ala Met Gln Trp 290 295 300 Asn Ser Thr Thr Phe His Gln Ala Leu Leu Asp Pro Arg Val Arg Gly 305 310 315 320 Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val 325 330 335 Pro Thr Thr Ala Ser Pro Ile Ser Ser Ile Phe Ser Arg Thr Gly Asp 340 345 350 Pro Ala Pro Asn Met Glu Asn Thr Thr Ser Gly Phe Leu Gly Pro Leu 355 360 365 Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile 370 375 380 Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly 385 390 395 400 Ala Pro Thr Cys Pro Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His 405 410 415 Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Cys 420 425 430 Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile 435 440 445 Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro 450 455 460 Leu Leu Pro Gly Thr Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr Cys 465 470 475 480 Thr Ile Pro Ala Gln Gly Thr Ser Met Phe Pro Ser Cys Cys Cys Thr 485 490 495 Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp 500 505 510 Ala Phe Ala Arg Phe Leu Trp Glu Trp Ala Ser Val Arg Phe Ser Trp 515 520 525 Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro 530 535 540 Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser 545 550 555 560 Leu Tyr Asn Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe 565 570 575 Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys 580 585 590 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 595 600 605 Phe Ser Arg Gly Ser Thr His Val Ser Trp Pro Lys Phe Ala Val Pro 610 615 620 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 625 630 635 640 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Ile Pro Leu His Pro Ala 645 650 655 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Pro Arg Tyr Val 660 665 670 Ala Arg Leu Ser Ser Thr Ser Arg Asn Ile Asn Tyr Gln His Gly Thr 675 680 685 Met Gln Asp Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 690 695 700 Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 705 710 715 720 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 725 730 735 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 740 745 750 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 755 760 765 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ser Ile 770 775 780 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 785 790 795 800 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 805 810 815 Trp Gly Thr Leu Pro Gln Glu His Ile Val Leu Lys Ile Lys Gln Cys 820 825 830 Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln 835 840 845 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 850 855 860 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala 865 870 875 880 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu 885 890 895 Asn Leu Tyr Pro Val Ala Arg Gln Arg Ser Gly Leu Cys Gln Val Phe 900 905 910 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Arg Arg 915 920 925 Met Arg Gly Thr Phe Val Ala Pro Leu Pro Ile His Thr Ala Glu Leu 930 935 940 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly 945 950 955 960 Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp 965 970 975 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 980 985 990 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 995 1000 1005 Leu Gly Leu Tyr Arg Pro Leu Leu His Leu Pro Phe Arg Pro Thr Thr 1010 1015 1020 Gly Arg Thr Ser Leu Tyr Ala Val Ser Pro Ser Val Pro Ser His Leu 1025 1030 1035 1040 Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro 1045 1050 1055 Pro <210> 2 <211> 1057 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 1_GL_epi <400> 2 Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys Ile 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg Lys 180 185 190 Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe Pro 195 200 205 Asp His Gln Leu Asp Pro Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp 210 215 220 Trp Asp Phe Asn Pro Asn Lys Asp His Trp Pro Glu Ala Asn Gln Val 225 230 235 240 Gly Ala Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Gly Leu 245 250 255 Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Thr Thr Val Pro Ala 260 265 270 Ala Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr 275 280 285 Pro Leu Ser Pro Pro Leu Arg Asp Thr His Pro Gln Ala Met Gln Trp 290 295 300 Asn Ser Thr Thr Phe His Gln Thr Leu Gln Asp Pro Arg Val Arg Gly 305 310 315 320 Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val 325 330 335 Pro Thr Thr Ala Ser Pro Ile Ser Ser Ile Phe Ser Arg Ile Gly Asp 340 345 350 Pro Ala Pro Asn Met Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu 355 360 365 Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile 370 375 380 Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly 385 390 395 400 Thr Pro Val Cys Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His 405 410 415 Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Cys 420 425 430 Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile 435 440 445 Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro 450 455 460 Leu Ile Pro Gly Ser Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr Cys 465 470 475 480 Thr Thr Pro Ala Gln Gly Thr Ser Met Phe Pro Ser Cys Cys Cys Thr 485 490 495 Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp 500 505 510 Ala Phe Ala Lys Phe Leu Trp Glu Trp Ala Ser Val Arg Phe Ser Trp 515 520 525 Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro 530 535 540 Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser 545 550 555 560 Leu Tyr Asn Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe 565 570 575 Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys 580 585 590 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 595 600 605 Phe Ser Arg Gly Asn Thr Arg Val Ser Trp Pro Lys Phe Ala Val Pro 610 615 620 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 625 630 635 640 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala 645 650 655 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val 660 665 670 Ala Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn His Gln His Gly Thr 675 680 685 Met Gln Asn Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 690 695 700 Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 705 710 715 720 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 725 730 735 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 740 745 750 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 755 760 765 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Tyr Thr Ala Val 770 775 780 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 785 790 795 800 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 805 810 815 Trp Gly Thr Leu Pro Gln Glu His Ile Val Gln Lys Ile Lys Gln Cys 820 825 830 Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln 835 840 845 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 850 855 860 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala 865 870 875 880 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu 885 890 895 Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe 900 905 910 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Gln Arg 915 920 925 Met Arg Gly Thr Phe Val Ala Pro Leu Pro Ile His Thr Ala Glu Leu 930 935 940 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly 945 950 955 960 Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp 965 970 975 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 980 985 990 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 995 1000 1005 Leu Gly Leu Tyr Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr 1010 1015 1020 Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu 1025 1030 1035 1040 Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro 1045 1050 1055 Pro <210> 3 <211> 1057 <212> PRT <213> artificial sequence <220> <223> synthetic sequence 2_CH_epi (a) <400> 3 Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys Ile 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg Gln 180 185 190 Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe Pro 195 200 205 Asp His Gln Leu Asp Pro Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp 210 215 220 Trp Asp Phe Asn Pro Asn Lys Asp His Trp Pro Glu Ala Asn Gln Val 225 230 235 240 Gly Ala Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Gly Leu 245 250 255 Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Thr Thr Val Pro Ala 260 265 270 Ala Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr 275 280 285 Pro Ile Ser Pro Pro Leu Arg Asp Ser His Pro Gln Ala Met Gln Trp 290 295 300 Asn Ser Thr Thr Phe His Gln Ala Leu Leu Asp Pro Arg Val Arg Gly 305 310 315 320 Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val 325 330 335 Pro Thr Thr Ala Ser Pro Ile Ser Ser Ile Phe Ser Arg Thr Gly Asp 340 345 350 Pro Ala Pro Asn Met Glu Asn Thr Thr Ser Gly Phe Leu Gly Pro Leu 355 360 365 Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile 370 375 380 Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly 385 390 395 400 Ala Pro Thr Cys Pro Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His 405 410 415 Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Cys 420 425 430 Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile 435 440 445 Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro 450 455 460 Leu Leu Pro Gly Thr Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr Cys 465 470 475 480 Thr Ile Pro Ala Gln Gly Thr Ser Met Phe Pro Ser Cys Cys Cys Thr 485 490 495 Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp 500 505 510 Ala Phe Ala Arg Phe Leu Trp Glu Trp Ala Ser Val Arg Phe Ser Trp 515 520 525 Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro 530 535 540 Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser 545 550 555 560 Leu Tyr Asn Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe 565 570 575 Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys 580 585 590 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 595 600 605 Phe Ser Arg Gly Ser Thr His Val Ser Trp Pro Lys Phe Ala Val Pro 610 615 620 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 625 630 635 640 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Ile Pro Leu His Pro Ala 645 650 655 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Pro Arg Tyr Val 660 665 670 Ala Arg Leu Ser Ser Thr Ser Arg Asn Ile Asn Tyr Gln His Gly Thr 675 680 685 Met Gln Asp Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 690 695 700 Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 705 710 715 720 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 725 730 735 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 740 745 750 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 755 760 765 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ser Ile 770 775 780 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 785 790 795 800 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 805 810 815 Trp Gly Thr Leu Pro Gln Glu His Ile Val Leu Lys Ile Lys Gln Cys 820 825 830 Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln 835 840 845 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 850 855 860 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala 865 870 875 880 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu 885 890 895 Asn Leu Tyr Pro Val Ala Arg Gln Arg Ser Gly Leu Cys Gln Val Phe 900 905 910 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Arg Arg 915 920 925 Met Arg Gly Thr Phe Val Ala Pro Leu Pro Ile His Thr Ala Glu Leu 930 935 940 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly 945 950 955 960 Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp 965 970 975 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 980 985 990 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 995 1000 1005 Leu Gly Leu Tyr Arg Pro Leu Leu His Leu Pro Phe Arg Pro Thr Thr 1010 1015 1020 Gly Arg Thr Ser Leu Tyr Ala Val Ser Pro Ser Val Pro Ser His Leu 1025 1030 1035 1040 Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro 1045 1050 1055 Pro <210> 4 <211> 1057 <212> PRT <213> artificial sequence <220> <223> synthetic sequence 2_CH_epi (b) <400> 4 Asp Ile Asp Thr Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Val Cys Trp Gly Glu Leu 50 55 60 Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Glu Leu Val Val Gly Tyr Val Asn Val Asn Met Gly Leu Lys Leu 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Thr Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Cys Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Lys 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg Lys 180 185 190 Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe Pro 195 200 205 Asp His Gln Leu Asp Pro Ala Phe Lys Ala Asn Ser Glu Asn Pro Asp 210 215 220 Trp Asp Leu Asn Pro His Lys Asp Asn Trp Pro Asp Ala Asn Lys Val 225 230 235 240 Gly Val Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Ser Leu 245 250 255 Leu Gly Trp Ser Pro Gln Ala Gln Gly Leu Leu Thr Thr Val Pro Val 260 265 270 Ala Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr 275 280 285 Pro Leu Ser Pro Pro Leu Arg Asp Thr His Pro Gln Ala Met Gln Trp 290 295 300 Asn Ser Thr Thr Phe His Gln Thr Leu Gln Asp Pro Arg Val Arg Ala 305 310 315 320 Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Ser Pro Ala 325 330 335 Gln Asn Thr Val Ser Ala Ile Ser Ser Ile Leu Ser Lys Thr Gly Asp 340 345 350 Pro Val Pro Asn Met Glu Asn Ile Ala Ser Gly Leu Leu Gly Pro Leu 355 360 365 Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Lys Ile Leu Thr Ile 370 375 380 Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly 385 390 395 400 Thr Pro Val Cys Leu Gly Gln Asn Ser Gln Ser Gln Ile Ser Ser His 405 410 415 Ser Pro Thr Cys Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Cys 420 425 430 Leu Arg Arg Phe Ile Ile Phe Leu Cys Ile Leu Leu Leu Cys Leu Ile 435 440 445 Phe Leu Leu Val Leu Val Asp Tyr Gln Gly Met Leu Pro Val Cys Pro 450 455 460 Leu Ile Pro Gly Ser Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr Cys 465 470 475 480 Thr Thr Pro Ala Gln Gly Thr Ser Met Tyr Pro Ser Cys Cys Cys Thr 485 490 495 Lys Pro Thr Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp 500 505 510 Ala Phe Ala Lys Tyr Leu Trp Glu Trp Ala Ser Val Arg Phe Ser Trp 515 520 525 Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro 530 535 540 Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Phe Trp Gly Pro Ser 545 550 555 560 Leu Tyr Asn Ile Leu Ser Pro Phe Met Pro Leu Leu Pro Ile Phe Phe 565 570 575 Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys 580 585 590 Asn Pro His Asn Thr Ala Glu Ser Arg Leu Val Val Asp Phe Ser Gln 595 600 605 Phe Ser Arg Gly Asn Thr Arg Val Ser Trp Pro Lys Phe Ala Val Pro 610 615 620 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 625 630 635 640 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala 645 650 655 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val 660 665 670 Ala Arg Leu Ser Ser Asn Ser Arg Ile Ile Asn Asn Gln His Arg Thr 675 680 685 Met Gln Asn Leu His Asn Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 690 695 700 Met Leu Leu Tyr Lys Thr Tyr Gly Arg Lys Leu His Leu Tyr Ser His 705 710 715 720 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 725 730 735 Pro Phe Leu Met Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 740 745 750 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Ile Asp Asp Val Val 755 760 765 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Tyr Ala Ala Val 770 775 780 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro His Lys Thr 785 790 795 800 Lys Arg Trp Gly Tyr Ser Leu His Phe Met Gly Tyr Val Ile Gly Cys 805 810 815 Trp Gly Thr Leu Pro Gln Glu His Ile Val Gln Lys Ile Lys Met Cys 820 825 830 Phe Arg Lys Leu Pro Val Asn Arg Pro Val Asp Trp Lys Val Cys Gln 835 840 845 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 850 855 860 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ala Lys Gln Ala 865 870 875 880 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Ser Lys Gln Tyr Leu 885 890 895 Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe 900 905 910 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Gln Arg 915 920 925 Met Arg Gly Thr Phe Val Ser Pro Leu Pro Ile His Thr Ala Glu Leu 930 935 940 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Asn Leu Ile Gly 945 950 955 960 Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp 965 970 975 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 980 985 990 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 995 1000 1005 Leu Gly Leu Tyr Arg Pro Leu Leu Arg Leu Leu Tyr Arg Pro Thr Thr 1010 1015 1020 Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu 1025 1030 1035 1040 Pro Val Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Lys Pro 1045 1050 1055 Pro <210> 5 <211> 1057 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 2_CHGL_epi Epi1 <400> 5 Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys Ile 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg Gln 180 185 190 Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe Pro 195 200 205 Asp His Gln Leu Asp Pro Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp 210 215 220 Trp Asp Phe Asn Pro Asn Lys Asp His Trp Pro Glu Ala Asn Gln Val 225 230 235 240 Gly Ala Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Gly Leu 245 250 255 Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Thr Thr Val Pro Ala 260 265 270 Ala Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr 275 280 285 Pro Ile Ser Pro Pro Leu Arg Asp Ser His Pro Gln Ala Met Gln Trp 290 295 300 Asn Ser Thr Thr Phe His Gln Ala Leu Leu Asp Pro Arg Val Arg Gly 305 310 315 320 Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val 325 330 335 Pro Thr Thr Ala Ser Pro Ile Ser Ser Ile Phe Ser Arg Thr Gly Asp 340 345 350 Pro Ala Pro Asn Met Glu Asn Thr Thr Ser Gly Phe Leu Gly Pro Leu 355 360 365 Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile 370 375 380 Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly 385 390 395 400 Ala Pro Thr Cys Pro Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His 405 410 415 Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Cys 420 425 430 Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile 435 440 445 Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro 450 455 460 Leu Leu Pro Gly Thr Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr Cys 465 470 475 480 Thr Ile Pro Ala Gln Gly Thr Ser Met Phe Pro Ser Cys Cys Cys Thr 485 490 495 Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp 500 505 510 Ala Phe Ala Arg Phe Leu Trp Glu Trp Ala Ser Val Arg Phe Ser Trp 515 520 525 Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro 530 535 540 Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser 545 550 555 560 Leu Tyr Asn Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe 565 570 575 Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys 580 585 590 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 595 600 605 Phe Ser Arg Gly Ser Thr His Val Ser Trp Pro Lys Phe Ala Val Pro 610 615 620 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 625 630 635 640 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Ile Pro Leu His Pro Ala 645 650 655 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Pro Arg Tyr Val 660 665 670 Ala Arg Leu Ser Ser Thr Ser Arg Asn Ile Asn Tyr Gln His Gly Thr 675 680 685 Met Gln Asp Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 690 695 700 Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 705 710 715 720 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 725 730 735 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 740 745 750 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 755 760 765 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ser Ile 770 775 780 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 785 790 795 800 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 805 810 815 Trp Gly Thr Leu Pro Gln Glu His Ile Val Leu Lys Ile Lys Gln Cys 820 825 830 Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln 835 840 845 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 850 855 860 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala 865 870 875 880 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu 885 890 895 Asn Leu Tyr Pro Val Ala Arg Gln Arg Ser Gly Leu Cys Gln Val Phe 900 905 910 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Arg Arg 915 920 925 Met Arg Gly Thr Phe Val Ala Pro Leu Pro Ile His Thr Ala Glu Leu 930 935 940 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly 945 950 955 960 Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp 965 970 975 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 980 985 990 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 995 1000 1005 Leu Gly Leu Tyr Arg Pro Leu Leu His Leu Pro Phe Arg Pro Thr Thr 1010 1015 1020 Gly Arg Thr Ser Leu Tyr Ala Val Ser Pro Ser Val Pro Ser His Leu 1025 1030 1035 1040 Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro 1045 1050 1055 Pro <210> 6 <211> 1048 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 2_CHGL_epi Epi2 <400> 6 Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Asp Ala Leu Glu Ser Pro Glu His Cys Thr 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Thr Leu Ala Thr Trp Val Gly Val Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys Phe 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Thr Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Asp Arg Gly Arg Ser Pro Arg Arg Arg 145 150 155 160 Thr Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg 165 170 175 Ser Lys Ser Arg Glu Ser Gln Cys Met Gly Gln Asn Leu Ser Thr Ser 180 185 190 Asn Pro Leu Gly Phe Phe Pro Asp His Gln Leu Asp Pro Ala Phe Arg 195 200 205 Ala Asn Thr Ala Asn Pro Asp Trp Asp Phe Asn Pro Asn Lys Asp Thr 210 215 220 Trp Pro Asp Ala Asn Lys Val Gly Ala Gly Ala Phe Gly Leu Gly Phe 225 230 235 240 Thr Pro Pro His Gly Gly Val Leu Gly Trp Ser Pro Gln Ala Gln Gly 245 250 255 Ile Leu Gln Thr Leu Pro Ala Asn Pro Pro Pro Ala Ser Thr Asn Arg 260 265 270 Gln Ser Gly Arg Gln Pro Thr Pro Leu Ser Pro Pro Leu Arg Asp Thr 275 280 285 His Pro Gln Ala Met Gln Trp Asn Ser Thr Thr Phe His Gln Thr Leu 290 295 300 Gln Asp Pro Arg Val Arg Ala Leu Tyr Phe Pro Ala Gly Gly Ser Ser 305 310 315 320 Ser Gly Thr Val Ser Pro Ala Gln Asn Thr Val Ser Ala Ile Ser Ser 325 330 335 Ile Leu Ser Lys Thr Gly Asp Pro Val Pro Asn Met Glu Asn Ile Ala 340 345 350 Ser Gly Leu Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe Leu 355 360 365 Leu Thr Lys Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp Thr 370 375 380 Ser Leu Asn Phe Leu Gly Gly Thr Thr Val Cys Leu Gly Gln Asn Ser 385 390 395 400 Gln Ser Pro Thr Ser Asn His Ser Pro Thr Ser Cys Pro Pro Thr Cys 405 410 415 Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe Ile Ile Phe Leu Cys 420 425 430 Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr Gln 435 440 445 Gly Met Leu Pro Val Cys Pro Leu Ile Pro Gly Ser Ser Thr Thr Ser 450 455 460 Thr Gly Pro Cys Arg Thr Cys Thr Thr Pro Ala Gln Gly Thr Ser Met 465 470 475 480 Tyr Pro Ser Cys Cys Cys Thr Lys Pro Thr Asp Gly Asn Cys Thr Cys 485 490 495 Ile Pro Ile Pro Ser Ser Trp Ala Phe Gly Lys Phe Leu Trp Glu Trp 500 505 510 Ala Ser Ala Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val Gln 515 520 525 Trp Phe Ala Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp Met 530 535 540 Met Trp Phe Trp Gly Pro Ser Leu Tyr Asn Ile Leu Ser Pro Phe Ile 545 550 555 560 Pro Leu Leu Pro Ile Phe Cys Tyr Leu Trp Val Ser Ile Val Thr Gly 565 570 575 Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr Ala Glu Ser Arg 580 585 590 Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Asn Thr Arg Val Ser 595 600 605 Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr Asn Leu Leu 610 615 620 Ser Ser Asp Leu Ser Trp Leu Ser Leu Asp Val Ser Ala Ala Phe Tyr 625 630 635 640 His Leu Pro Leu His Pro Ala Ala Met Pro His Leu Leu Val Gly Ser 645 650 655 Ser Gly Leu Ser Arg Tyr Val Ala Arg Leu Ser Ser Asn Ser Arg Ile 660 665 670 Phe Asn His Gln His Gly Thr Met Gln Asn Leu His Asp Ser Cys Ser 675 680 685 Arg Asn Leu Tyr Val Ser Leu Met Leu Leu Tyr Lys Thr Tyr Gly Arg 690 695 700 Lys Leu His Leu Tyr Ser His Pro Ile Ile Met Gly Phe Arg Lys Ile 705 710 715 720 Pro Met Gly Val Gly Leu Ser Pro Phe Leu Met Ala Gln Phe Thr Ser 725 730 735 Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His Cys Leu Ala Phe 740 745 750 Ser Tyr Met Asp Asp Leu Val Leu Gly Ala Lys Ser Val Gln His Leu 755 760 765 Glu Ser Leu Phe Thr Ala Val Thr Asn Phe Leu Leu Ser Leu Gly Ile 770 775 780 His Leu Asn Pro His Lys Thr Lys Arg Trp Gly Tyr Ser Leu His Phe 785 790 795 800 Met Gly Tyr Val Ile Gly Cys Tyr Gly Ser Leu Pro Gln Asp His Ile 805 810 815 Ile Gln Lys Ile Lys Glu Cys Phe Arg Lys Leu Pro Val Asn Arg Pro 820 825 830 Ile Asp Trp Lys Val Cys Gln Arg Ile Val Gly Leu Leu Gly Phe Ala 835 840 845 Ala Pro Phe Thr Gln Cys Gly Tyr Pro Ala Leu Met Pro Leu Tyr Ala 850 855 860 Cys Ile Gln Ala Lys Gln Ala Phe Thr Phe Ser Pro Thr Tyr Lys Ala 865 870 875 880 Phe Leu Ser Lys Gln Tyr Met Asn Leu Tyr Pro Val Ala Arg Gln Arg 885 890 895 Pro Gly Leu Cys Gln Val Phe Ala Asp Ala Thr Pro Thr Gly Trp Gly 900 905 910 Leu Ala Ile Gly His Gln Arg Met Arg Gly Thr Phe Val Ser Pro Leu 915 920 925 Pro Ile His Thr Ala Glu Leu Leu Ala Ala Cys Phe Ala Arg Ser Arg 930 935 940 Ser Gly Ala Asn Ile Leu Gly Thr Asp Asn Ser Val Val Leu Ser Arg 945 950 955 960 Lys Tyr Thr Ser Phe Pro Trp Leu Leu Gly Cys Thr Ala Asn Trp Ile 965 970 975 Leu Arg Gly Thr Ser Phe Val Tyr Val Pro Ser Ala Leu Asn Pro Ala 980 985 990 Asp Asp Pro Ser Arg Gly Arg Leu Gly Leu Ser Arg Pro Leu Leu Arg 995 1000 1005 Leu Pro Phe Arg Pro Thr Thr Gly Arg Thr Ser Leu Tyr Ala Asp Ser 1010 1015 1020 Pro Ser Val Pro Ser His Leu Pro Val Arg Val His Phe Ala Ser Pro 1025 1030 1035 1040 Leu His Val Ala Trp Lys Pro Pro 1045 <210> 7 <211> 1063 <212> PRT <213> artificial sequence <220> <223> Synthesis Epi3 <400> 7 Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu 1 5 10 15 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp Leu Leu Asp 20 25 30 Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys 35 40 45 Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 50 55 60 Leu Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala 65 70 75 80 Ser Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys 85 90 95 Ile Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg 100 105 110 Glu Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr 115 120 125 Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro 130 135 140 Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr 145 150 155 160 Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser 165 170 175 Gln Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg 180 185 190 Gln Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe 195 200 205 Pro Asp His Gln Leu Asp Pro Ala Phe Gly Ala Asn Ser Asn Asn Pro 210 215 220 Asp Trp Asp Phe Asn Pro Asn Lys Asp His Trp Pro Glu Ala Asn Gln 225 230 235 240 Val Gly Ala Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Gly 245 250 255 Leu Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Thr Thr Val Pro 260 265 270 Ala Ala Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro 275 280 285 Thr Pro Ile Ser Pro Pro Leu Arg Asp Ser His Pro Gln Ala Met Gln 290 295 300 Trp Asn Ser Thr Thr Phe His Gln Ala Leu Leu Asp Pro Arg Val Arg 305 310 315 320 Gly Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro 325 330 335 Val Pro Thr Thr Ala Ser Pro Ile Ser Ser Ile Phe Ser Arg Thr Gly 340 345 350 Asp Pro Ala Pro Asn Met Glu Asn Thr Ala Ser Gly Phe Leu Gly Pro 355 360 365 Leu Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr 370 375 380 Ile Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly 385 390 395 400 Gly Ala Pro Thr Cys Pro Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn 405 410 415 His Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met 420 425 430 Cys Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu 435 440 445 Ile Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys 450 455 460 Pro Leu Leu Pro Gly Thr Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr 465 470 475 480 Cys Thr Ile Pro Ala Gln Gly Thr Ser Met Phe Pro Ser Cys Cys Cys 485 490 495 Thr Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser 500 505 510 Trp Ala Phe Ala Arg Phe Leu Trp Glu Trp Ala Ser Val Arg Phe Ser 515 520 525 Trp Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser 530 535 540 Pro Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro 545 550 555 560 Ser Leu Tyr Asn Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe 565 570 575 Phe Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp 580 585 590 Lys Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser 595 600 605 Gln Phe Ser Arg Gly Ser Thr His Val Ser Trp Pro Lys Phe Ala Val 610 615 620 Pro Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp 625 630 635 640 Leu Ser Leu Asp Val Ser Ala Ala Phe Tyr His Ile Pro Leu His Pro 645 650 655 Ala Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Pro Arg Tyr 660 665 670 Val Ala Arg Leu Ser Ser Thr Ser Arg Asn Ile Asn Tyr Gln His Gly 675 680 685 Thr Met Gln Asp Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser 690 695 700 Leu Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser 705 710 715 720 His Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu 725 730 735 Ser Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val 740 745 750 Arg Arg Ala Phe Pro His Cys Leu Ala Phe Ser Val Val Leu Gly Ala 755 760 765 Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ser Ile Thr Asn Phe 770 775 780 Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr Lys Arg Trp 785 790 795 800 Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser Trp Gly Thr 805 810 815 Leu Pro Gln Glu His Ile Val Leu Lys Ile Lys Gln Cys Phe Arg Lys 820 825 830 Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln Arg Ile Val 835 840 845 Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly Tyr Pro Ala 850 855 860 Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala Phe Thr Phe 865 870 875 880 Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu Asn Leu Tyr 885 890 895 Pro Val Ala Arg Gln Arg Ser Gly Leu Cys Gln Val Phe Ala Ala Ala 900 905 910 Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Arg Arg Met Arg Gly 915 920 925 Thr Phe Val Ala Pro Leu Pro Ile His Thr Ala Ala Leu Leu Ala Ala 930 935 940 Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly Thr Val Asn 945 950 955 960 Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp Leu Leu Gly 965 970 975 Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val Tyr Val Pro 980 985 990 Ser Ala Leu Asn Pro Ala Ala Asp Pro Ser Arg Gly Arg Leu Gly Leu 995 1000 1005 Tyr Arg Pro Leu Leu His Leu Pro Phe Arg Pro Thr Thr Gly Arg Thr 1010 1015 1020 Ser Leu Tyr Ala Val Ser Pro Ser Val Pro Ser His Leu Pro Asp Arg 1025 1030 1035 1040 Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro Pro Ile Pro 1045 1050 1055 Asn Pro Leu Leu Gly Leu Asp 1060 <210> 8 <211> 1054 <212> PRT <213> artificial sequence <220> <223> Synthesis Epi4 <400> 8 Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu 1 5 10 15 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp 20 25 30 Thr Ala Ser Ala Leu Tyr Arg Asp Ala Leu Glu Ser Pro Glu His Cys 35 40 45 Thr Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 50 55 60 Leu Met Thr Leu Ala Thr Trp Val Gly Val Asn Leu Glu Asp Pro Ala 65 70 75 80 Ser Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys 85 90 95 Phe Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg 100 105 110 Glu Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr 115 120 125 Pro Thr Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro 130 135 140 Glu Thr Thr Val Val Arg Arg Arg Asp Arg Gly Arg Ser Pro Arg Arg 145 150 155 160 Arg Thr Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg 165 170 175 Arg Ser Lys Ser Arg Glu Ser Gln Cys Met Gly Gln Asn Leu Ser Thr 180 185 190 Ser Asn Pro Leu Gly Phe Phe Pro Asp His Gln Leu Asp Pro Ala Phe 195 200 205 Arg Ala Asn Thr Ala Asn Pro Asp Trp Asp Phe Asn Pro Asn Lys Asp 210 215 220 Thr Trp Pro Asp Ala Asn Lys Val Gly Ala Gly Ala Phe Gly Leu Gly 225 230 235 240 Phe Thr Pro Pro His Gly Gly Val Leu Gly Trp Ser Pro Gln Ala Gln 245 250 255 Gly Ile Leu Gln Thr Leu Pro Ala Asn Pro Pro Pro Ala Ser Thr Asn 260 265 270 Arg Gln Ser Gly Arg Gln Pro Thr Pro Leu Ser Pro Pro Leu Arg Asp 275 280 285 Thr His Pro Gln Ala Met Gln Trp Asn Ser Thr Thr Phe His Gln Thr 290 295 300 Leu Gln Asp Pro Arg Val Arg Ala Leu Tyr Phe Pro Ala Gly Gly Ser 305 310 315 320 Ser Ser Gly Thr Val Ser Pro Ala Gln Asn Thr Val Ser Ala Ile Ser 325 330 335 Ser Ile Leu Ser Lys Thr Gly Asp Pro Val Pro Asn Met Glu Asn Ile 340 345 350 Ala Ser Gly Leu Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe 355 360 365 Leu Leu Thr Lys Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp 370 375 380 Thr Ser Leu Asn Phe Leu Gly Gly Thr Thr Val Cys Leu Gly Gln Asn 385 390 395 400 Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr Ser Cys Pro Pro Thr 405 410 415 Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe Ile Ile Phe Leu 420 425 430 Cys Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr 435 440 445 Gln Gly Met Leu Pro Val Cys Pro Leu Ile Pro Gly Ser Ser Thr Thr 450 455 460 Ser Thr Gly Pro Cys Arg Thr Cys Thr Thr Pro Ala Gln Gly Thr Ser 465 470 475 480 Met Tyr Pro Ser Cys Cys Cys Thr Lys Pro Thr Asp Gly Asn Cys Thr 485 490 495 Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Gly Lys Phe Leu Trp Glu 500 505 510 Trp Ala Ser Ala Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val 515 520 525 Gln Trp Phe Ala Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp 530 535 540 Met Met Trp Phe Trp Gly Pro Ser Leu Tyr Asn Ile Leu Ser Pro Phe 545 550 555 560 Ile Pro Leu Leu Pro Ile Phe Cys Tyr Leu Trp Val Ser Ile Val Thr 565 570 575 Gly Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr Ala Glu Ser 580 585 590 Arg Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Asn Thr Arg Val 595 600 605 Ser Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr Asn Leu 610 615 620 Leu Ser Ser Asp Leu Ser Trp Leu Ser Leu Asp Val Ser Ala Ala Phe 625 630 635 640 Tyr His Leu Pro Leu His Pro Ala Ala Met Pro His Leu Leu Val Gly 645 650 655 Ser Ser Gly Leu Ser Arg Tyr Val Ala Arg Leu Ser Ser Asn Ser Arg 660 665 670 Ile Phe Asn His Gln His Gly Thr Met Gln Asn Leu His Asp Ser Cys 675 680 685 Ser Arg Asn Leu Tyr Val Ser Leu Met Leu Leu Tyr Lys Thr Tyr Gly 690 695 700 Arg Lys Leu His Leu Tyr Ser His Pro Ile Ile Met Gly Phe Arg Lys 705 710 715 720 Ile Pro Met Gly Val Gly Leu Ser Pro Phe Leu Met Ala Gln Phe Thr 725 730 735 Ser Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His Cys Leu Ala 740 745 750 Phe Ser Leu Val Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu 755 760 765 Phe Thr Ala Val Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn 770 775 780 Pro His Lys Thr Lys Arg Trp Gly Tyr Ser Leu His Phe Met Gly Tyr 785 790 795 800 Val Ile Gly Cys Tyr Gly Ser Leu Pro Gln Asp His Ile Ile Gln Lys 805 810 815 Ile Lys Glu Cys Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp 820 825 830 Lys Val Cys Gln Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe 835 840 845 Thr Gln Cys Gly Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln 850 855 860 Ala Lys Gln Ala Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Ser 865 870 875 880 Lys Gln Tyr Met Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu 885 890 895 Cys Gln Val Phe Ala Ala Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile 900 905 910 Gly His Gln Arg Met Arg Gly Thr Phe Val Ser Pro Leu Pro Ile His 915 920 925 Thr Ala Ala Leu Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala 930 935 940 Asn Ile Leu Gly Thr Val Asn Ser Val Val Leu Ser Arg Lys Tyr Thr 945 950 955 960 Ser Phe Pro Trp Leu Leu Gly Cys Thr Ala Asn Trp Ile Leu Arg Gly 965 970 975 Thr Ser Phe Val Tyr Val Pro Ser Ala Leu Asn Pro Ala Ala Asp Pro 980 985 990 Ser Arg Gly Arg Leu Gly Leu Ser Arg Pro Leu Leu Arg Leu Pro Phe 995 1000 1005 Arg Pro Thr Thr Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val 1010 1015 1020 Pro Ser His Leu Pro Val Arg Val His Phe Ala Ser Pro Leu His Val 1025 1030 1035 1040 Ala Trp Lys Pro Pro Tyr Pro Tyr Asp Val Pro Asp Tyr Ala<00者01568>1045 1050 <210> 9 <211> 1039 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence 3_GL_epi Rare 15.1 <400> 9 Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu Ser Phe 1 5 10 15 Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp Leu Leu Asp Thr Ala 20 25 30 Ser Ala Leu Tyr Arg Asp Ala Leu Glu Ser Pro Glu His Cys Thr Pro 35 40 45 His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu Met 50 55 60 Thr Leu Ala Thr Trp Val Gly Val Asn Leu Glu Asp Pro Ala Ser Arg 65 70 75 80 Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys Phe Arg 85 90 95 Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu Thr 100 105 110 Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro Thr 115 120 125 Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu Thr 130 135 140 Thr Val Val Arg Arg Arg Asp Arg Gly Arg Ser Pro Arg Arg Arg Thr 145 150 155 160 Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Arg Glu Ser Gln Cys Met 165 170 175 Gly Gln Asn Leu Ser Thr Ser Asn Pro Leu Gly Phe Phe Pro Asp His 180 185 190 Gln Leu Asp Pro Ala Phe Lys Ala Asn Ser Glu Asn Pro Asp Trp Asp 195 200 205 Leu Asn Pro His Lys Asp Asn Trp Pro Asp Ala Asn Lys Val Gly Ala 210 215 220 Gly Ala Phe Gly Leu Gly Phe Thr Pro Pro His Gly Gly Val Leu Gly 225 230 235 240 Trp Ser Pro Gln Ala Gln Gly Ile Leu Gln Thr Leu Pro Ala Asn Pro 245 250 255 Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr Pro Leu 260 265 270 Ser Pro Pro Leu Arg Asp Thr His Pro Gln Ala Met Gln Trp Asn Ser 275 280 285 Thr Thr Phe His Gln Ala Leu Leu Asp Pro Arg Val Arg Gly Leu Tyr 290 295 300 Leu Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Ser Pro Ala Gln Asn 305 310 315 320 Thr Val Ser Ala Ile Ser Ser Ile Leu Ser Lys Thr Gly Asp Pro Val 325 330 335 Pro Asn Met Glu Asn Ile Ala Ser Gly Leu Leu Gly Pro Leu Leu Val 340 345 350 Leu Gln Ala Gly Phe Phe Leu Leu Thr Lys Ile Leu Thr Ile Pro Gln 355 360 365 Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly Ala Pro 370 375 380 Thr Cys Pro Gly Gln Asn Ser Gln Ser Pro Thr Ser Ser His Ser Pro 385 390 395 400 Thr Ser Cys Pro Pro Thr Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg 405 410 415 Arg Phe Ile Ile Phe Leu Cys Ile Leu Leu Leu Cys Leu Ile Phe Leu 420 425 430 Leu Val Leu Val Asp Tyr Gln Gly Met Leu Pro Val Cys Pro Leu Leu 435 440 445 Pro Gly Thr Ser Thr Thr Ser Thr Gly Pro Cys Arg Thr Cys Thr Thr 450 455 460 Pro Ala Gln Gly Thr Ser Met Tyr Pro Ser Cys Cys Cys Thr Lys Pro 465 470 475 480 Thr Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe 485 490 495 Gly Lys Phe Leu Trp Glu Trp Ala Ser Ala Arg Phe Ser Trp Leu Ser 500 505 510 Leu Leu Val Pro Phe Val Gln Trp Phe Ala Gly Leu Ser Pro Thr Val 515 520 525 Trp Leu Ser Ala Ile Trp Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr 530 535 540 Ser Ile Leu Ser Pro Phe Ile Pro Leu Leu Pro Ile Phe Phe Cys Leu 545 550 555 560 Trp Ala Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys Asn Pro 565 570 575 His Asn Thr Ala Glu Ser Arg Leu Val Val Asp Phe Ser Gln Phe Ser 580 585 590 Arg Gly Ser Thr His Val Ser Trp Pro Lys Phe Ala Val Pro Asn Leu 595 600 605 Gln Ser Leu Thr Asn Leu Leu Ser Ser Asp Leu Ser Trp Leu Ser Leu 610 615 620 Asp Val Ser Ala Ala Phe Tyr His Ile Pro Leu His Pro Ala Ala Met 625 630 635 640 Pro His Leu Leu Ile Gly Ser Ser Gly Leu Ser Arg Tyr Val Ala Arg 645 650 655 Leu Ser Ser Thr Ser Arg Asn Ile Asn Tyr Gln His Gly Thr Met Gln 660 665 670 Asp Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu Met Leu 675 680 685 Leu Tyr Lys Thr Tyr Gly Arg Lys Leu His Leu Tyr Ser His Pro Ile 690 695 700 Ile Met Gly Phe Arg Lys Ile Pro Met Gly Leu Gly Leu Ser Pro Phe 705 710 715 720 Leu Met Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg Arg Ala 725 730 735 Phe Pro His Cys Leu Ala Phe Ser Tyr Ile Asp Asp Val Val Leu Gly 740 745 750 Ala Lys Thr Val Gln His Leu Glu Ser Leu Phe Thr Ser Ile Thr Asn 755 760 765 Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro His Lys Thr Lys Arg 770 775 780 Trp Gly Tyr Ser Leu His Phe Met Gly Tyr Val Ile Gly Cys Tyr Gly 785 790 795 800 Ser Leu Pro Gln Asp His Ile Ile Gln Lys Ile Lys Glu Cys Phe Arg 805 810 815 Lys Leu Pro Val Asn Arg Pro Val Asp Trp Lys Val Cys Gln Arg Ile 820 825 830 Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly Tyr Pro 835 840 845 Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ala Lys Gln Ala Phe Thr 850 855 860 Phe Ser Pro Pro Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Met Asn Leu 865 870 875 880 Tyr Pro Val Ala Arg Gln Arg Ser Gly Leu Cys Gln Val Phe Ala Asp 885 890 895 Ala Thr Pro Thr Gly Trp Gly Leu Val Met Gly His Gln Arg Met Arg 900 905 910 Gly Thr Phe Val Ser Pro Leu Pro Ile His Thr Ala Glu Leu Leu Ala 915 920 925 Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Asn Ile Leu Gly Thr Asp 930 935 940 Asn Ser Val Val Leu Ala Arg Lys Tyr Thr Ser Phe Pro Trp Leu Leu 945 950 955 960 Gly Cys Thr Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val Tyr Val 965 970 975 Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg Leu Gly 980 985 990 Leu Ser Arg Pro Leu Leu Arg Leu Pro Phe Gln Pro Thr Thr Gly Arg 995 1000 1005 Thr Ser Leu Tyr Ala Val Ser Pro Ser Val Pro Ser His Leu Pro Val 1010 1015 1020 Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Lys Pro Pro 1025 1030 1035 <210> 10 <211> 1049 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence 3_GL_epi Rare 15.2 <400> 10 Asp Ile Asp Thr Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ala Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ala Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Thr 35 40 45 Pro Asn His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Asp Leu 50 55 60 Met Thr Leu Ala Thr Trp Val Gly Asn Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Asp Leu Val Val Asn Tyr Val Asn Thr Asn Met Gly Leu Lys Ile 85 90 95 Arg Gln Ile Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Asp 100 105 110 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Gln Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Asn Thr Val Val Arg Arg Arg Gly Arg Thr Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Arg Glu Pro Gln Cys Met Gly 165 170 175 Gly Trp Ser Ser Lys Pro Arg Gln Gly Met Gly Thr Asn Leu Ser Val 180 185 190 Pro Asn Pro Leu Gly Phe Leu Pro Asp His Gln Leu Asp Pro Ala Phe 195 200 205 Arg Ala Asn Thr Ala Asn Pro Asp Trp Asp Phe Asn Pro Asn Lys Asp 210 215 220 His Trp Pro Glu Ala Asn Gln Val Gly Ala Gly Ala Phe Gly Pro Gly 225 230 235 240 Phe Thr Pro Pro His Gly Asn Leu Leu Gly Trp Ser Pro Gln Ala Gln 245 250 255 Gly Leu Leu Thr Thr Val Pro Val Ala Pro Pro Pro Ala Ser Thr Asn 260 265 270 Arg Gln Leu Gly Arg Lys Pro Thr Pro Leu Ser Pro Pro Leu Arg Asn 275 280 285 Thr His Pro Gln Ala Met Gln Trp Asn Ser Thr Ala Phe His Gln Ala 290 295 300 Leu Gln Asp Pro Arg Val Arg Ala Leu Tyr Phe Pro Ala Gly Gly Ser 305 310 315 320 Thr Ser Gly Thr Val Asn Pro Val Pro Thr Thr Val Ser Pro Ile Ser 325 330 335 Ser Ile Phe Ser Arg Thr Gly Asp Pro Ala Pro Asn Met Glu Asn Thr 340 345 350 Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe 355 360 365 Ser Leu Thr Lys Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp 370 375 380 Thr Ser Leu Asn Phe Leu Gly Gly Thr Pro Val Cys Leu Gly Gln Asn 385 390 395 400 Ser Gln Ser Gln Ile Ser Ser His Ser Pro Thr Cys Cys Pro Pro Ile 405 410 415 Cys Pro Gly Tyr Arg Trp Met Tyr Leu Arg Arg Phe Ile Ile Phe Leu 420 425 430 Phe Ile Leu Leu Leu Cys Leu Thr Phe Leu Leu Val Leu Leu Asp Tyr 435 440 445 Arg Gly Met Leu Pro Val Cys Pro Leu Ile Pro Gly Ser Thr Thr Thr 450 455 460 Ser Thr Gly Pro Cys Lys Thr Cys Thr Ile Pro Ala Gln Gly Thr Ser 465 470 475 480 Met Phe Pro Ser Cys Cys Cys Ser Lys Pro Ser Asp Gly Asn Cys Thr 485 490 495 Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Ala Arg Phe Leu Trp Glu 500 505 510 Trp Ala Ser Val Arg Phe Ser Trp Leu Ser Leu Leu Val Gln Phe Val 515 520 525 Gln Trp Cys Val Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp 530 535 540 Met Met Trp Phe Trp Gly Pro Ser Leu Tyr Asn Ile Leu Ser Pro Phe 545 550 555 560 Met Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp Val Tyr Ile Ile Thr 565 570 575 Gly Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr Glu Glu Ser 580 585 590 Arg Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Asn Tyr Arg Val 595 600 605 Ser Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr Asn Leu 610 615 620 Leu Ser Ser Asn Leu Ser Trp Ile Ser Leu Asp Val Ser Ala Ala Phe 625 630 635 640 Tyr His Leu Pro Leu His Pro Ala Ala Met Pro His Leu Leu Val Gly 645 650 655 Ser Ser Gly Leu Pro Arg Tyr Val Ala Arg Leu Ser Ser Asn Ser Arg 660 665 670 Ile Ile Asn Asn Gln His Arg Thr Met Gln Asn Leu His Asn Ser Cys 675 680 685 Ser Arg Asn Leu Tyr Val Ser Leu Leu Leu Leu Tyr Gln Thr Phe Gly 690 695 700 Arg Lys Leu His Leu Tyr Ser His Pro Ile Val Leu Gly Phe Arg Lys 705 710 715 720 Ile Pro Met Gly Val Gly Leu Ser Pro Phe Leu Leu Ala Gln Phe Thr 725 730 735 Ser Ala Leu Cys Ser Val Val Arg Arg Ala Phe Pro His Cys Leu Ala 740 745 750 Phe Ser Tyr Met Asp Asp Leu Val Leu Gly Ala Lys Ser Val Gln His 755 760 765 Leu Glu Ser Leu Tyr Ala Ala Val Thr Asn Phe Leu Leu Ser Val Gly 770 775 780 Ile His Leu Asn Thr Ser Lys Thr Lys Arg Trp Gly Tyr Ser Leu Asn 785 790 795 800 Phe Met Gly Tyr Ile Ile Gly Ser Trp Gly Thr Leu Pro Gln Asp His 805 810 815 Ile Val Gln Lys Ile Lys His Cys Phe Arg Lys Leu Pro Val Asn Arg 820 825 830 Pro Ile Asp Trp Lys Val Cys Gln Arg Leu Val Gly Leu Leu Gly Phe 835 840 845 Ala Ala Pro Phe Thr Gln Cys Gly Tyr Pro Ala Leu Lys Pro Leu Tyr 850 855 860 Ala Cys Ile Gln Ser Arg Gln Ala Phe Thr Phe Ser Pro Thr Tyr Lys 865 870 875 880 Ala Phe Leu Ser Lys Gln Tyr Leu Asn Leu Tyr Pro Val Ala Arg Gln 885 890 895 Arg Ser Gly Val Cys Gln Val Phe Ala Asp Ala Thr Pro Thr Gly Trp 900 905 910 Gly Leu Ala Ile Gly His Arg Arg Met Arg Gly Thr Phe Val Ala Pro 915 920 925 Leu Pro Ile His Thr Ala Glu Leu Leu Ala Ala Cys Phe Ala Arg Ser 930 935 940 Arg Ser Gly Ala Asn Leu Ile Gly Thr Asp Asn Ser Val Val Leu Ser 945 950 955 960 Arg Lys Tyr Thr Ser Tyr Pro Trp Leu Leu Gly Cys Ala Ala Asn Trp 965 970 975 Ile Leu Arg Gly Thr Ser Phe Val Tyr Val Pro Ser Ala Leu Asn Pro 980 985 990 Ala Asp Asp Pro Ser Arg Gly Arg Leu Gly Ile Tyr Arg Pro Leu Leu 995 1000 1005 Arg Leu Pro Tyr Arg Pro Thr Thr Gly Arg Thr Ser Leu Tyr Ala Ala 1010 1015 1020 Ser Pro Ser Val Pro Ser His Leu Pro Ala Arg Val His Phe Ala Ser 1025 1030 1035 1040 Pro Leu His Val Thr Trp Lys Pro Pro 1045 <210> 11 <211> 1057 <212> PRT <213> Synthetic Sequence <220> <223> Synthetic Sequence 3_GL_epi Rare 15.3 <400> 11<0​​​​​​​​​​​​Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys Ile 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Lys 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg Lys 180 185 190 Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe Pro 195 200 205 Asp His Gln Leu Asp Pro Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp 210 215 220 Trp Asp Phe Asn Pro Asn Lys Asp Thr Trp Pro Asp Ala Asn Lys Val 225 230 235 240 Gly Val Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Gly Leu 245 250 255 Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Thr Thr Val Pro Ala 260 265 270 Ala Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr 275 280 285 Pro Ile Ser Pro Pro Leu Arg Asp Ser His Pro Gln Ala Met Gln Trp 290 295 300 Asn Ser Thr Thr Phe His Gln Thr Leu Gln Asp Pro Arg Val Arg Gly 305 310 315 320 Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val 325 330 335 Pro Thr Thr Ala Ser Pro Ile Ser Ser Ile Phe Ser Arg Ile Gly Asp 340 345 350 Pro Ala Leu Asn Met Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu 355 360 365 Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile 370 375 380 Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly 385 390 395 400 Thr Thr Val Cys Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His 405 410 415 Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Cys 420 425 430 Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile 435 440 445 Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro 450 455 460 Leu Ile Pro Gly Ser Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr Cys 465 470 475 480 Thr Thr Pro Ala Gln Gly Asn Ser Met Phe Pro Ser Cys Cys Cys Thr 485 490 495 Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp 500 505 510 Ala Phe Ala Lys Tyr Leu Trp Glu Trp Ala Ser Val Arg Phe Ser Trp 515 520 525 Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro 530 535 540 Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser 545 550 555 560 Leu Tyr Asn Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe 565 570 575 Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys 580 585 590 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 595 600 605 Phe Ser Arg Gly Asn Thr Arg Val Ser Trp Pro Lys Phe Ala Val Pro 610 615 620 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 625 630 635 640 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala 645 650 655 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val 660 665 670 Ala Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn His Gln His Gly Thr 675 680 685 Met Gln Asn Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 690 695 700 Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 705 710 715 720 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 725 730 735 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 740 745 750 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 755 760 765 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ala Val 770 775 780 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 785 790 795 800 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 805 810 815 Trp Gly Thr Leu Pro Gln Glu His Ile Val Gln Lys Ile Lys Gln Cys 820 825 830 Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln 835 840 845 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 850 855 860 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala 865 870 875 880 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu 885 890 895 Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe 900 905 910 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Gln Arg 915 920 925 Met Arg Gly Thr Phe Leu Ala Pro Leu Pro Ile His Thr Ala Glu Leu 930 935 940 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly 945 950 955 960 Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp 965 970 975 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 980 985 990 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 995 1000 1005 Leu Gly Leu Tyr Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr 1010 1015 1020 Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu 1025 1030 1035 1040 Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro 1045 1050 1055 Pro <210> 12 <211> 1046 <212> PRT <213> Hepatitis B virus <400> 12 Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Thr Leu Ala Thr Trp Val Gly Gly Asn Leu Glu Asp Pro Ile Ser 65 70 75 80 Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys Phe 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gln Asn Leu Ser Thr Ser Asn Pro 180 185 190 Leu Gly Phe Phe Pro Asp His Gln Leu Asp Pro Ala Phe Arg Ala Asn 195 200 205 Thr Ala Asn Pro Asp Trp Asp Phe Asn Pro Asn Lys Asp Thr Trp Pro 210 215 220 Asp Ala Asn Lys Val Gly Ala Gly Ala Phe Gly Leu Gly Phe Thr Pro 225 230 235 240 Pro His Gly Gly Leu Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu 245 250 255 Gln Thr Leu Pro Ala Asn Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser 260 265 270 Gly Arg Gln Pro Thr Pro Leu Ser Pro Pro Leu Arg Asn Thr His Pro 275 280 285 Gln Ala Met Gln Trp Asn Ser Thr Thr Phe His Gln Thr Leu Gln Asp 290 295 300 Pro Arg Val Arg Gly Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly 305 310 315 320 Thr Val Asn Pro Val Pro Thr Thr Val Ser Pro Ile Ser Ser Ile Phe 325 330 335 Ser Arg Ile Gly Asp Pro Ala Leu Asn Met Glu Asn Ile Thr Ser Gly 340 345 350 Phe Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe Leu Leu Thr 355 360 365 Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu 370 375 380 Asn Phe Leu Gly Gly Thr Thr Val Cys Leu Gly Gln Asn Ser Gln Ser 385 390 395 400 Pro Thr Ser Asn His Ser Pro Thr Ser Cys Pro Pro Thr Cys Pro Gly 405 410 415 Tyr Arg Trp Met Cys Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu 420 425 430 Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met 435 440 445 Leu Pro Val Cys Pro Leu Ile Pro Gly Ser Ser Thr Thr Ser Thr Gly 450 455 460 Pro Cys Arg Thr Cys Thr Thr Pro Ala Gln Gly Thr Ser Met Tyr Pro 465 470 475 480 Ser Cys Cys Cys Thr Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro 485 490 495 Ile Pro Ser Ser Trp Ala Phe Gly Lys Phe Leu Trp Glu Trp Ala Ser 500 505 510 Ala Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe 515 520 525 Val Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp Met Met Trp 530 535 540 Tyr Trp Gly Pro Ser Leu Tyr Ser Ile Leu Ser Pro Phe Leu Pro Leu 545 550 555 560 Leu Pro Ile Phe Phe Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val 565 570 575 Phe Leu Val Asp Lys Asn Pro His Asn Thr Ala Glu Ser Arg Leu Val 580 585 590 Val Asp Phe Ser Gln Phe Ser Arg Gly Asn Tyr Arg Val Ser Trp Pro 595 600 605 Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser 610 615 620 Asn Leu Ser Trp Leu Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu 625 630 635 640 Pro Leu His Pro Ala Ala Met Pro His Leu Leu Val Gly Ser Ser Gly 645 650 655 Leu Ser Arg Tyr Val Ala Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn 660 665 670 Tyr Gln His Gly Thr Met Gln Asn Leu His Asp Ser Cys Ser Arg Asn 675 680 685 Leu Tyr Val Ser Leu Leu Leu Leu Tyr Gln Thr Phe Gly Arg Lys Leu 690 695 700 His Leu Tyr Ser His Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met 705 710 715 720 Gly Val Gly Leu Ser Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile 725 730 735 Cys Ser Val Val Arg Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr 740 745 750 Met Asp Asp Val Val Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser 755 760 765 Leu Phe Thr Ala Val Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu 770 775 780 Asn Pro Asn Lys Thr Lys Arg Trp Gly Tyr Ser Leu His Phe Met Gly 785 790 795 800 Tyr Val Ile Gly Cys Tyr Gly Ser Leu Pro Gln Asp His Ile Ile Gln 805 810 815 Lys Ile Lys Glu Cys Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp 820 825 830 Trp Lys Val Cys Gln Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro 835 840 845 Phe Thr Gln Cys Gly Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile 850 855 860 Gln Ser Lys Gln Ala Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu 865 870 875 880 Cys Lys Gln Tyr Leu Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly 885 890 895 Leu Cys Gln Val Phe Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Val 900 905 910 Met Gly His Gln Arg Met Arg Gly Thr Phe Leu Ala Pro Leu Pro Ile 915 920 925 His Thr Ala Glu Leu Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly 930 935 940 Ala Asn Ile Leu Gly Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr 945 950 955 960 Thr Ser Phe Pro Trp Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg 965 970 975 Gly Thr Ser Phe Val Tyr Val Pro Ser Ala Leu Asn Pro Thr Asp Asp 980 985 990 Pro Ser Arg Gly Arg Leu Gly Leu Ser Arg Pro Leu Leu Arg Leu Pro 995 1000 1005 Phe Arg Pro Thr Thr Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser 1010 1015 1020 Val Pro Ser His Leu Pro Asp Arg Val His Phe Ala Ser Pro Leu His 1025 1030 1035 1040 Val Ala Trp Arg Pro Pro 1045 <210> 13 <211> 1057 <212> PRT <213> Hepatitis B virus <400> 13 Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu Ser 1 5 10 15 Phe Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp Leu Leu Asp Thr 20 25 30 Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser 35 40 45 Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu 50 55 60 Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala Ser 65 70 75 80 Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys Ile 85 90 95 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 100 105 110 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 115 120 125 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 130 135 140 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 145 150 155 160 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Gln 165 170 175 Ser Arg Glu Ser Gln Cys Met Gly Gly Trp Ser Ser Lys Pro Arg Lys 180 185 190 Gly Met Gly Thr Asn Leu Ser Val Pro Asn Pro Leu Gly Phe Phe Pro 195 200 205 Asp His Gln Leu Asp Pro Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp 210 215 220 Trp Asp Phe Asn Pro Asn Lys Asp His Trp Pro Glu Ala Asn Gln Val 225 230 235 240 Gly Ala Gly Ala Phe Gly Pro Gly Phe Thr Pro Pro His Gly Gly Leu 245 250 255 Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Thr Thr Val Pro Ala 260 265 270 Ala Pro Pro Pro Ala Ala Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr 275 280 285 Pro Ile Ser Pro Pro Leu Arg Asp Ser His Pro Gln Ala Met Gln Trp 290 295 300 Asn Ser Thr Thr Phe His Gln Ala Leu Leu Asp Pro Arg Val Arg Gly 305 310 315 320 Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val 325 330 335 Pro Thr Thr Ala Ser Pro Ile Ser Ser Ile Phe Ser Arg Ser Gly Asp 340 345 350 Pro Ala Pro Asn Met Glu Asn Thr Thr Ser Gly Phe Leu Gly Pro Leu 355 360 365 Leu Val Leu Gln Ala Val Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile 370 375 380 Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Glu 385 390 395 400 Ala Pro Ala Cys Pro Gly Gln Asn Ser Gln Ser Pro Thr Ser Ser His 405 410 415 Ser Pro Thr Ser Cys Pro Pro Ile Cys Pro Gly Tyr Arg Trp Met Cys 420 425 430 Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile 435 440 445 Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro 450 455 460 Leu Leu Pro Gly Thr Ser Thr Thr Ser Thr Gly Pro Cys Lys Thr Cys 465 470 475 480 Thr Ile Pro Ala Gln Gly Thr Ser Met Phe Pro Ser Cys Cys Cys Thr 485 490 495 Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp 500 505 510 Ala Phe Ala Arg Phe Leu Trp Glu Trp Ala Ser Val Arg Phe Ser Trp 515 520 525 Leu Ser Leu Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro 530 535 540 Thr Val Trp Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser 545 550 555 560 Leu Tyr Asn Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Leu Phe Phe 565 570 575 Cys Leu Trp Val Tyr Ile Val Thr Gly Gly Val Phe Leu Val Asp Lys 580 585 590 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 595 600 605 Phe Ser Arg Gly Ser Thr Arg Val Ser Trp Pro Lys Phe Ala Val Pro 610 615 620 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 625 630 635 640 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala 645 650 655 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Pro Arg Tyr Val 660 665 670 Ala Arg Leu Ser Ser Ala Ser Arg Asn Ile Asn Tyr Gln His Gly Thr 675 680 685 Met Gln Asp Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 690 695 700 Met Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 705 710 715 720 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 725 730 735 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 740 745 750 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 755 760 765 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ser Ile 770 775 780 Thr Thr Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 785 790 795 800 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 805 810 815 Trp Gly Thr Leu Pro Gln Glu His Ile Val Leu Lys Ile Lys Gln Cys 820 825 830 Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln 835 840 845 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 850 855 860 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ala Lys Gln Ala 865 870 875 880 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Arg Thr Gln Tyr Leu 885 890 895 Asn Leu Tyr Pro Val Ala Arg Gln Arg Ser Gly Leu Cys Gln Val Phe 900 905 910 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Met Gly His Gln Arg 915 920 925 Met Arg Gly Thr Phe Leu Ala Pro Leu Pro Ile His Thr Ala Glu Leu 930 935 940 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly 945 950 955 960 Thr Asp Asn Ser Val Val Leu Ala Arg Lys Tyr Thr Ser Phe Pro Trp 965 970 975 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 980 985 990 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 995 1000 1005 Leu Gly Leu Tyr Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr 1010 1015 1020 Gly Arg Thr Ser Leu Tyr Ala Val Ser Pro Ser Val Pro Ser His Leu 1025 1030 1035 1040 Pro Val Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro 1045 1050 1055 Pro <210> 14 <211> 1404 <212> PRT <213> artificial sequence <220> <223> Synthesis of HBV epigraph <400> 14 Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu 1 5 10 15 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp 20 25 30 Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys 35 40 45 Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 50 55 60 Leu Met Thr Leu Ala Thr Trp Val Gly Gly Asn Leu Glu Asp Pro Ile 65 70 75 80 Ser Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys 85 90 95 Phe Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg 100 105 110 Glu Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr 115 120 125 Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro 130 135 140 Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr 145 150 155 160 Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser 165 170 175 Lys Ser Arg Glu Ser Gln Cys Met Gly Gln Asn Leu Ser Thr Ser Asn 180 185 190 Pro Leu Gly Phe Phe Pro Asp His Gln Leu Asp Pro Ala Phe Arg Ala 195 200 205 Asn Thr Ala Asn Pro Asp Trp Asp Phe Asn Pro Asn Lys Asp Thr Trp 210 215 220 Pro Asp Ala Asn Lys Val Gly Ala Gly Ala Phe Gly Leu Gly Phe Thr 225 230 235 240 Pro Pro His Gly Gly Leu Leu Gly Trp Ser Pro Gln Ala Gln Gly Ile 245 250 255 Leu Gln Thr Leu Pro Ala Asn Pro Pro Pro Ala Ser Thr Asn Arg Gln 260 265 270 Ser Gly Arg Gln Pro Thr Pro Leu Ser Pro Pro Leu Arg Asn Thr His 275 280 285 Pro Gln Ala Met Gln Trp Asn Ser Thr Thr Phe His Gln Thr Leu Gln 290 295 300 Asp Pro Arg Val Arg Gly Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser 305 310 315 320 Gly Thr Val Asn Pro Val Pro Thr Thr Val Ser His Ile Ser Ser Ile 325 330 335 Phe Ser Arg Ile Gly Asp Pro Ala Leu Asn Met Glu Asn Ile Thr Ser 340 345 350 Gly Phe Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe Leu Leu 355 360 365 Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp Thr Ser 370 375 380 Leu Asn Phe Leu Gly Gly Thr Thr Val Cys Leu Gly Gln Asn Ser Gln 385 390 395 400 Ser Pro Thr Ser Asn His Ser Pro Thr Ser Cys Pro Pro Thr Cys Pro 405 410 415 Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe Ile Ile Phe Leu Phe Ile 420 425 430 Leu Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr Gln Gly 435 440 445 Met Leu Pro Val Cys Pro Leu Ile Pro Gly Ser Ser Thr Thr Ser Thr 450 455 460 Gly Pro Cys Arg Thr Cys Thr Thr Pro Ala Gln Gly Thr Ser Met Tyr 465 470 475 480 Pro Ser Cys Cys Cys Thr Lys Pro Ser Asp Gly Asn Cys Thr Cys Ile 485 490 495 Pro Ile Pro Ser Ser Trp Ala Phe Gly Lys Phe Leu Trp Glu Trp Ala 500 505 510 Ser Ala Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val Gln Trp 515 520 525 Phe Val Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp Met Met 530 535 540 Trp Tyr Trp Gly Pro Ser Leu Tyr Ser Ile Leu Ser Pro Phe Leu Pro 545 550 555 560 Leu Leu Pro Ile Phe Phe Cys Leu Trp Val Tyr Ile Met Pro Leu Ser 565 570 575 Tyr Gln His Phe Arg Arg Leu Leu Leu Leu Asp Asp Glu Ala Gly Pro 580 585 590 Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp Glu Gly Leu Asn Arg Arg 595 600 605 Val Ala Glu Asp Leu Asn Leu Gly Asn Leu Asn Val Ser Ile Pro Trp 610 615 620 Thr His Lys Val Gly Asn Phe Thr Gly Leu Tyr Ser Ser Thr Val Pro 625 630 635 640 Val Phe Asn Pro His Trp Lys Thr Pro Ser Phe Pro Asn Ile His Leu 645 650 655 His Gln Asp Ile Ile Lys Lys Cys Glu Gln Phe Val Gly Pro Leu Thr 660 665 670 Val Asn Glu Lys Arg Arg Leu Gln Leu Ile Met Pro Ala Arg Phe Tyr 675 680 685 Pro Asn Val Thr Lys Tyr Leu Pro Leu Asp Lys Gly Ile Lys Pro Tyr 690 695 700 Tyr Pro Glu His Leu Val Asn His Tyr Phe Gln Thr Arg His Tyr Leu 705 710 715 720 His Thr Leu Trp Lys Ala Gly Ile Leu Tyr Lys Arg Glu Thr Thr His 725 730 735 Ser Ala Ser Phe Cys Gly Ser Pro Tyr Ser Trp Glu Gln Glu Leu Gln 740 745 750 His Gly Ala Glu Ser Phe His Gln Gln Ser Ser Gly Ile Leu Ser Arg 755 760 765 Pro Pro Val Gly Ser Ser Leu Gln Ser Lys His Arg Lys Ser Arg Leu 770 775 780 Gly Leu Gln Ser Gln Gln Gly His Leu Ala Arg Arg Gln Gln Gly Arg 785 790 795 800 Ser Trp Ser Ile Arg Ala Gly Ile His Pro Thr Ala Arg Arg Pro Phe 805 810 815 Gly Val Glu Pro Ser Gly Ser Gly His Thr Thr Asn Leu Ala Ser Lys 820 825 830 Ser Ala Ser Cys Leu Tyr Gln Ser Pro Val Arg Lys Ala Ala Tyr Pro 835 840 845 Ala Val Ser Thr Phe Glu Lys His Ser Ser Ser Gly His Ala Val Glu 850 855 860 Leu His Asn Leu Pro Pro Asn Ser Ala Arg Ser Gln Ser Glu Arg Pro 865 870 875 880 Val Phe Pro Cys Trp Trp Leu Gln Phe Arg Asn Ser Lys Pro Cys Ser 885 890 895 Asp Tyr Cys Leu Ser His Ile Val Asn Leu Leu Glu Asp Trp Gly Pro 900 905 910 Cys Ala Glu His Gly Glu His His Ile Arg Ile Pro Arg Thr Pro Ala 915 920 925 Arg Val Thr Gly Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr 930 935 940 Ala Glu Ser Arg Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Asn 945 950 955 960 Tyr Arg Val Ser Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu 965 970 975 Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu Ser Leu Asp Val Ser 980 985 990 Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala Ala Met Pro His Leu 995 1000 1005 Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val Ala Arg Leu Ser Ser 1010 1015 1020 Asn Ser Arg Ile Phe Asn His Gln His Gly Thr Met Gln Asn Leu His 1025 1030 1035 1040 Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu Leu Leu Leu Tyr Gln 1045 1050 1055 Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His Pro Ile Ile Leu Gly 1060 1065 1070 Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser Pro Phe Leu Leu Ala 1075 1080 1085 Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His 1090 1095 1100 Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val Leu Gly Ala Lys Ser 1105 1110 1115 1120 Val Gln His Leu Glu Ser Leu Phe Thr Ala Val Thr Asn Phe Leu Leu 1125 1130 1135 Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr Lys Arg Trp Gly Tyr 1140 1145 1150 Ser Leu His Phe Met Gly Tyr Val Ile Gly Cys Tyr Gly Ser Leu Pro 1155 1160 1165 Gln Asp His Ile Ile Gln Lys Ile Lys Glu Cys Phe Arg Lys Leu Pro 1170 1175 1180 Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln Arg Ile Val Gly Leu 1185 1190 1195 1200 Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly Tyr Pro Ala Leu Met 1205 1210 1215 Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala Phe Thr Phe Ser Pro 1220 1225 1230 Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu Asn Leu Tyr Pro Val 1235 1240 1245 Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe Ala Asp Ala Thr Pro 1250 1255 1260 Thr Gly Trp Gly Leu Val Met Gly His Gln Arg Met Arg Gly Thr Phe 1265 1270 1275 1280 Leu Ala Pro Leu Pro Ile His Thr Ala Glu Leu Leu Ala Ala Cys Phe 1285 1290 1295 Ala Arg Ser Arg Ser Gly Ala Asn Ile Leu Gly Thr Asp Asn Ser Val 1300 1305 1310 Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp Leu Leu Gly Cys Ala 1315 1320 1325 Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val Tyr Val Pro Ser Ala 1330 1335 1340 Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg Leu Gly Leu Ser Arg 1345 1350 1355 1360 Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr Gly Arg Thr Ser Leu 1365 1370 1375 Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu Pro Asp Arg Val His 1380 1385 1390 Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro Pro 1395 1400 <210> 15 <211> 1396 <212> PRT <213> Artificial sequence <220> <223> Synthetic sequence HBV epigraph subtype D Sequence_SPC Reordering <400> 15 Met Gly Gln Asn Leu Ser Thr Ser Asn Pro Leu Gly Phe Phe Pro Asp 1 5 10 15 His Gln Leu Asp Pro Ala Phe Arg Ala Asn Thr Ala Asn Pro Asp Trp 20 25 30 Asp Phe Asn Pro Asn Lys Asp Thr Trp Pro Asp Ala Asn Lys Val Gly 35 40 45 Ala Gly Ala Phe Gly Leu Gly Phe Thr Pro Pro His Gly Gly Leu Leu 50 55 60 Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Gln Thr Leu Pro Ala Asn 65 70 75 80 Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr Pro 85 90 95 Leu Ser Pro Pro Leu Arg Asn Thr His Pro Gln Ala Gln Trp Asn Ser 100 105 110 Thr Thr Phe His Gln Thr Leu Gln Asp Pro Arg Val Arg Gly Leu Tyr 115 120 125 Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val Pro Thr 130 135 140 Thr Val Ser His Ile Ser Ser Ile Phe Ser Arg Ile Gly Asp Pro Ala 145 150 155 160 Leu Asn Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu 165 170 175 Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser 180 185 190 Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly Thr Thr Val 195 200 205 Cys Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr 210 215 220 Ser Cys Pro Pro Thr Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg 225 230 235 240 Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu 245 250 255 Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro Leu Ile Pro 260 265 270 Gly Ser Ser Thr Thr Ser Thr Gly Pro Cys Arg Thr Cys Thr Thr Pro 275 280 285 Ala Gln Gly Thr Ser Met Tyr Pro Ser Cys Cys Cys Thr Lys Pro Ser 290 295 300 Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Gly 305 310 315 320 Lys Phe Leu Trp Glu Trp Ala Ser Ala Arg Phe Ser Trp Leu Ser Leu 325 330 335 Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp 340 345 350 Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Ser 355 360 365 Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp 370 375 380 Val Tyr Ile Pro Leu Ser Tyr Gln His Phe Arg Arg Leu Leu Leu Leu 385 390 395 400 Asp Asp Glu Ala Gly Pro Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp 405 410 415 Glu Gly Leu Asn Arg Arg Val Ala Glu Asp Leu Asn Leu Gly Asn Leu 420 425 430 Asn Val Ser Ile Pro Trp Thr His Lys Val Gly Asn Phe Thr Gly Leu 435 440 445 Tyr Ser Ser Thr Val Pro Val Phe Asn Pro His Trp Lys Thr Pro Ser 450 455 460 Phe Pro Asn Ile His Leu His Gln Asp Ile Ile Lys Lys Cys Glu Gln 465 470 475 480 Phe Val Gly Pro Leu Thr Val Asn Glu Lys Arg Arg Leu Gln Leu Ile 485 490 495 Met Pro Ala Arg Phe Tyr Pro Asn Val Thr Lys Tyr Leu Pro Leu Asp 500 505 510 Lys Gly Ile Lys Pro Tyr Tyr Pro Glu His Leu Val Asn His Tyr Phe 515 520 525 Gln Thr Arg His Tyr Leu His Thr Leu Trp Lys Ala Gly Ile Leu Tyr 530 535 540 Lys Arg Glu Thr Thr His Ser Ala Ser Phe Cys Gly Ser Pro Tyr Ser 545 550 555 560 Trp Glu Gln Glu Leu Gln His Gly Ala Glu Ser Phe His Gln Gln Ser 565 570 575 Ser Gly Ile Leu Ser Arg Pro Pro Val Gly Ser Ser Leu Gln Ser Lys 580 585 590 His Arg Lys Ser Arg Leu Gly Leu Gln Ser Gln Gln Gly His Leu Ala 595 600 605 Arg Arg Gln Gln Gly Arg Ser Trp Ser Ile Arg Ala Gly Ile His Pro 610 615 620 Thr Ala Arg Arg Pro Phe Gly Val Glu Pro Ser Gly Ser Gly His Thr 625 630 635 640 Thr Asn Leu Ala Ser Lys Ser Ala Ser Cys Leu Tyr Gln Ser Pro Val 645 650 655 Arg Lys Ala Ala Tyr Pro Ala Val Ser Thr Phe Glu Lys His Ser Ser 660 665 670 Ser Gly His Ala Val Glu Leu His Asn Leu Pro Pro Asn Ser Ala Arg 675 680 685 Ser Gln Ser Glu Arg Pro Val Phe Pro Cys Trp Trp Leu Gln Phe Arg 690 695 700 Asn Ser Lys Pro Cys Ser Asp Tyr Cys Leu Ser His Ile Val Asn Leu 705 710 715 720 Leu Glu Asp Trp Gly Pro Cys Ala Glu His Gly Glu His His Ile Arg 725 730 735 Ile Pro Arg Thr Pro Ala Arg Val Thr Gly Gly Val Phe Leu Val Asp 740 745 750 Lys Asn Pro His Asn Thr Ala Glu Ser Arg Leu Val Val Asp Phe Ser 755 760 765 Gln Phe Ser Arg Gly Asn Tyr Arg Val Ser Trp Pro Lys Phe Ala Val 770 775 780 Pro Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp 785 790 795 800 Leu Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro 805 810 815 Ala Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr 820 825 830 Val Ala Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn His Gln His Gly 835 840 845 Thr Met Gln Asn Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser 850 855 860 Leu Leu Leu Leu Tyr Gln Thr Phe Gly Arg Lys Leu His Leu Tyr Ser 865 870 875 880 His Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu 885 890 895 Ser Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val 900 905 910 Arg Arg Ala Phe Pro His Cys Leu Ala Phe Ser Val Val Leu Gly Ala 915 920 925 Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ala Val Thr Asn Phe 930 935 940 Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr Lys Arg Trp 945 950 955 960 Gly Tyr Ser Leu His Phe Met Gly Tyr Val Ile Gly Cys Tyr Gly Ser 965 970 975 Leu Pro Gln Asp His Ile Ile Gln Lys Ile Lys Glu Cys Phe Arg Lys 980 985 990 Leo Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln Arg Ile Val 995 1000 1005 Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly Tyr Pro Ala 1010 1015 1020 Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala Phe Thr Phe 1025 1030 1035 1040 Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu Asn Leu Tyr 1045 1050 1055 Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe Ala Ala Ala 1060 1065 1070 Thr Pro Thr Gly Trp Gly Leu Val Met Gly His Gln Arg Met Arg Gly 1075 1080 1085 Thr Phe Leu Ala Pro Leu Pro Ile His Thr Ala Ala Leu Leu Ala Ala 1090 1095 1100 Cys Phe Ala Arg Ser Arg Ser Gly Ala Asn Ile Leu Gly Thr Val Asn 1105 1110 1115 1120 Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp Leu Leu Gly 1125 1130 1135 Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val Tyr Val Pro 1140 1145 1150 Ser Ala Leu Asn Pro Ala Ala Asp Pro Ser Arg Gly Arg Leu Gly Leu 1155 1160 1165 Ser Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr Gly Arg Thr 1170 1175 1180 Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu Pro Asp Arg 1185 1190 1195 1200 Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro Pro Asp Ile 1205 1210 1215 Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu Ser Phe Leu 1220 1225 1230 Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr Ala Ser 1235 1240 1245 Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser Pro His 1250 1255 1260 His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu Met Thr 1265 1270 1275 1280 Leu Ala Thr Trp Val Gly Gly Asn Leu Glu Asp Pro Ile Ser Arg Asp 1285 1290 1295 Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys Phe Arg Gln 1300 1305 1310 Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu Thr Val 1315 1320 1325 Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro Pro Ala 1330 1335 1340 Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu Thr Thr 1345 1350 1355 1360 Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro Ser Pro 1365 1370 1375 Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Lys Ser Arg 1380 1385 1390 Glu Ser Gln Cys 1395 <210> 16 <211> 183 <212> PRT <213> artificial sequence <220> <223> Synthesis of EpiD core(C) protein <400> 16 Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu 1 5 10 15 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp 20 25 30 Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys 35 40 45 Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 50 55 60 Leu Met Thr Leu Ala Thr Trp Val Gly Gly Asn Leu Glu Asp Pro Ile 65 70 75 80 Ser Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys 85 90 95 Phe Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg 100 105 110 Glu Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr 115 120 125 Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro 130 135 140 Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr 145 150 155 160 Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser 165 170 175 Lys Ser Arg Glu Ser Gln Cys 180 <210> 17 <211> 108 <212> PRT <213> Artificial sequence <220> <223> Synthetic sequence EpiD PreS1 domain <400> 17 Met Gly Gln Asn Leu Ser Thr Ser Asn Pro Leu Gly Phe Phe Pro Asp 1 5 10 15 His Gln Leu Asp Pro Ala Phe Arg Ala Asn Thr Ala Asn Pro Asp Trp 20 25 30 Asp Phe Asn Pro Asn Lys Asp Thr Trp Pro Asp Ala Asn Lys Val Gly 35 40 45 Ala Gly Ala Phe Gly Leu Gly Phe Thr Pro Pro His Gly Gly Leu Leu 50 55 60 Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Gln Thr Leu Pro Ala Asn 65 70 75 80 Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr Pro 85 90 95 Leu Ser Pro Pro Leu Arg Asn Thr His Pro Gln Ala 100 105 <210> 18 <211> 55 <212> PRT <213> Artificial sequence <220> <223> Synthetic sequence EpiD PreS2 domain <400> 18 Met Gln Trp Asn Ser Thr Thr Phe His Gln Thr Leu Gln Asp Pro Arg 1 5 10 15 Val Arg Gly Leu Tyr Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val 20 25 30 Asn Pro Val Pro Thr Thr Val Ser Pro Ile Ser Ser Ile Phe Ser Arg 35 40 45 Ile Gly Asp Pro Ala Leu Asn 50 55 <210> 19 <211> 226 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence EpiD Surface Antigen (S) <400> 19 Met Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu Gln 1 5 10 15 Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu 20 25 30 Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly Thr Thr Val Cys 35 40 45 Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr Ser 50 55 60 Cys Pro Pro Thr Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe 65 70 75 80 Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val 85 90 95 Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro Leu Ile Pro Gly 100 105 110 Ser Ser Thr Thr Ser Thr Gly Pro Cys Arg Thr Cys Thr Thr Pro Ala 115 120 125 Gln Gly Thr Ser Met Tyr Pro Ser Cys Cys Cys Thr Lys Pro Ser Asp 130 135 140 Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Gly Lys 145 150 155 160 Phe Leu Trp Glu Trp Ala Ser Ala Arg Phe Ser Trp Leu Ser Leu Leu 165 170 175 Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp Leu 180 185 190 Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Ser Ile 195 200 205 Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp Val 210 215 220 Tyr Island 225 <210> 20 <211> 832 <212> PRT <213> artificial sequence <220> <223> Synthesis EpiD Polymerase (P) <400> 20 Met Pro Leu Ser Tyr Gln His Phe Arg Arg Leu Leu Leu Leu Asp Asp 1 5 10 15 Glu Ala Gly Pro Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp Glu Gly 20 25 30 Leu Asn Arg Arg Val Ala Glu Asp Leu Asn Leu Gly Asn Leu Asn Val 35 40 45 Ser Ile Pro Trp Thr His Lys Val Gly Asn Phe Thr Gly Leu Tyr Ser 50 55 60 Ser Thr Val Pro Val Phe Asn Pro His Trp Lys Thr Pro Ser Phe Pro 65 70 75 80 Asn Ile His Leu His Gln Asp Ile Ile Lys Lys Cys Glu Gln Phe Val 85 90 95 Gly Pro Leu Thr Val Asn Glu Lys Arg Arg Leu Gln Leu Ile Met Pro 100 105 110 Ala Arg Phe Tyr Pro Asn Val Thr Lys Tyr Leu Pro Leu Asp Lys Gly 115 120 125 Ile Lys Pro Tyr Tyr Pro Glu His Leu Val Asn His Tyr Phe Gln Thr 130 135 140 Arg His Tyr Leu His Thr Leu Trp Lys Ala Gly Ile Leu Tyr Lys Arg 145 150 155 160 Glu Thr Thr His Ser Ala Ser Phe Cys Gly Ser Pro Tyr Ser Trp Glu 165 170 175 Gln Glu Leu Gln His Gly Ala Glu Ser Phe His Gln Gln Ser Ser Gly 180 185 190 Ile Leu Ser Arg Pro Pro Val Gly Ser Ser Leu Gln Ser Lys His Arg 195 200 205 Lys Ser Arg Leu Gly Leu Gln Ser Gln Gln Gly His Leu Ala Arg Arg 210 215 220 Gln Gln Gly Arg Ser Trp Ser Ile Arg Ala Gly Ile His Pro Thr Ala 225 230 235 240 Arg Arg Pro Phe Gly Val Glu Pro Ser Gly Ser Gly His Thr Thr Asn 245 250 255 Leu Ala Ser Lys Ser Ala Ser Cys Leu Tyr Gln Ser Pro Val Arg Lys 260 265 270 Ala Ala Tyr Pro Ala Val Ser Thr Phe Glu Lys His Ser Ser Ser Gly 275 280 285 His Ala Val Glu Leu His Asn Leu Pro Pro Asn Ser Ala Arg Ser Gln 290 295 300 Ser Glu Arg Pro Val Phe Pro Cys Trp Trp Leu Gln Phe Arg Asn Ser 305 310 315 320 Lys Pro Cys Ser Asp Tyr Cys Leu Ser His Ile Val Asn Leu Leu Glu 325 330 335 Asp Trp Gly Pro Cys Ala Glu His Gly Glu His His Ile Arg Ile Pro 340 345 350 Arg Thr Pro Ala Arg Val Thr Gly Gly Val Phe Leu Val Asp Lys Asn 355 360 365 Pro His Asn Thr Ala Glu Ser Arg Leu Val Val Asp Phe Ser Gln Phe 370 375 380 Ser Arg Gly Asn Tyr Arg Val Ser Trp Pro Lys Phe Ala Val Pro Asn 385 390 395 400 Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu Ser 405 410 415 Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala Ala Ala 420 425 430 Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val Ala 435 440 445 Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn His Gln His Gly Thr Met 450 455 460 Gln Asn Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu Leu 465 470 475 480 Leu Leu Tyr Gln Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His Pro 485 490 495 Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser Pro 500 505 510 Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg Arg 515 520 525 Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val Leu 530 535 540 Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ala Val Thr 545 550 555 560 Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr Lys 565 570 575 Arg Trp Gly Tyr Ser Leu His Phe Met Gly Tyr Val Ile Gly Cys Tyr 580 585 590 Gly Ser Leu Pro Gln Asp His Ile Ile Gln Lys Ile Lys Glu Cys Phe 595 600 605 Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln Arg 610 615 620 Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly Tyr 625 630 635 640 Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala Phe 645 650 655 Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu Asn 660 665 670 Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe Ala 675 680 685 Asp Ala Thr Pro Thr Gly Trp Gly Leu Val Met Gly His Gln Arg Met 690 695 700 Arg Gly Thr Phe Leu Ala Pro Leu Pro Ile His Thr Ala Glu Leu Leu 705 710 715 720 Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Asn Ile Leu Gly Thr 725 730 735 Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp Leu 740 745 750 Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val Tyr 755 760 765 Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg Leu 770 775 780 Gly Leu Ser Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr Gly 785 790 795 800 Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu Pro 805 810 815 Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro Pro 820 825 830 <210> twenty one <211> 183 <212> PRT <213> Artificial sequence <220> <223> Conserved regions of the synthetic sequence of EpiD Core (C); shortened <400> twenty one Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Thr Val Glu Leu Leu 1 5 10 15 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp 20 25 30 Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys 35 40 45 Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 50 55 60 Leu Met Thr Leu Ala Thr Trp Val Gly Gly Asn Leu Glu Asp Pro Ile 65 70 75 80 Ser Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys 85 90 95 Phe Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg 100 105 110 Glu Thr Val Ile Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr 115 120 125 Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro 130 135 140 Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr 145 150 155 160 Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser 165 170 175 Lys Ser Arg Glu Ser Gln Cys 180 <210> twenty two <211> 475 <212> PRT <213> Artificial sequence <220> <223> Conserved regions of the synthetic sequence of EpiD Polymerase (P) <400> twenty two Val Thr Gly Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr Ala 1 5 10 15 Glu Ser Arg Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Asn Tyr 20 25 30 Arg Val Ser Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr 35 40 45 Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu Ser Leu Asp Val Ser Ala 50 55 60 Ala Phe Tyr His Leu Pro Leu His Pro Ala Ala Met Pro His Leu Leu 65 70 75 80 Val Gly Ser Ser Gly Leu Ser Arg Tyr Val Ala Arg Leu Ser Ser Asn 85 90 95 Ser Arg Ile Phe Asn His Gln His Gly Thr Met Gln Asn Leu His Asp 100 105 110 Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu Leu Leu Leu Tyr Gln Thr 115 120 125 Phe Gly Arg Lys Leu His Leu Tyr Ser His Pro Ile Ile Leu Gly Phe 130 135 140 Arg Lys Ile Pro Met Gly Val Gly Leu Ser Pro Phe Leu Leu Ala Gln 145 150 155 160 Phe Thr Ser Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His Cys 165 170 175 Leu Ala Phe Ser Tyr Met Asp Asp Val Val Leu Gly Ala Lys Ser Val 180 185 190 Gln His Leu Glu Ser Leu Phe Thr Ala Val Thr Asn Phe Leu Leu Ser 195 200 205 Leu Gly Ile His Leu Asn Pro Asn Lys Thr Lys Arg Trp Gly Tyr Ser 210 215 220 Leu His Phe Met Gly Tyr Val Ile Gly Cys Tyr Gly Ser Leu Pro Gln 225 230 235 240 Asp His Ile Ile Gln Lys Ile Lys Glu Cys Phe Arg Lys Leu Pro Val 245 250 255 Asn Arg Pro Ile Asp Trp Lys Val Cys Gln Arg Ile Val Gly Leu Leu 260 265 270 Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly Tyr Pro Ala Leu Met Pro 275 280 285 Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala Phe Thr Phe Ser Pro Thr 290 295 300 Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu Asn Leu Tyr Pro Val Ala 305 310 315 320 Arg Gln Arg Pro Gly Leu Cys Gln Val Phe Ala Asp Ala Thr Pro Thr 325 330 335 Gly Trp Gly Leu Val Met Gly His Gln Arg Met Arg Gly Thr Phe Leu 340 345 350 Ala Pro Leu Pro Ile His Thr Ala Glu Leu Leu Ala Ala Cys Phe Ala 355 360 365 Arg Ser Arg Ser Gly Ala Asn Ile Leu Gly Thr Asp Asn Ser Val Val 370 375 380 Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp Leu Leu Gly Cys Ala Ala 385 390 395 400 Asn Trp Ile Leu Arg Gly Thr Ser Phe Val Tyr Val Pro Ser Ala Leu 405 410 415 Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg Leu Gly Leu Ser Arg Pro 420 425 430 Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr Gly Arg Thr Ser Leu Tyr 435 440 445 Ala Asp Ser Pro Ser Val Pro Ser His Leu Pro Asp Arg Val His Phe 450 455 460 Ala Ser Pro Leu His Val Ala Trp Arg Pro Pro 465 470 475 <210> 23 <211> 471 <212> PRT <213> artificial sequence <220> <223> Conserved regions of the synthesis sequence of EpiD mutant polymerase (P) It has mutations from Table 1 <400> twenty three Val Thr Gly Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr Ala 1 5 10 15 Glu Ser Arg Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Asn Tyr 20 25 30 Arg Val Ser Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr 35 40 45 Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu Ser Leu Asp Val Ser Ala 50 55 60 Ala Phe Tyr His Leu Pro Leu His Pro Ala Ala Met Pro His Leu Leu 65 70 75 80 Val Gly Ser Ser Gly Leu Ser Arg Tyr Val Ala Arg Leu Ser Ser Asn 85 90 95 Ser Arg Ile Phe Asn His Gln His Gly Thr Met Gln Asn Leu His Asp 100 105 110 Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu Leu Leu Leu Tyr Gln Thr 115 120 125 Phe Gly Arg Lys Leu His Leu Tyr Ser His Pro Ile Ile Leu Gly Phe 130 135 140 Arg Lys Ile Pro Met Gly Val Gly Leu Ser Pro Phe Leu Leu Ala Gln 145 150 155 160 Phe Thr Ser Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His Cys 165 170 175 Leu Ala Phe Ser Val Val Leu Gly Ala Lys Ser Val Gln His Leu Glu 180 185 190 Ser Leu Phe Thr Ala Val Thr Asn Phe Leu Leu Ser Leu Gly Ile His 195 200 205 Leu Asn Pro Asn Lys Thr Lys Arg Trp Gly Tyr Ser Leu His Phe Met 210 215 220 Gly Tyr Val Ile Gly Cys Tyr Gly Ser Leu Pro Gln Asp His Ile Ile 225 230 235 240 Gln Lys Ile Lys Glu Cys Phe Arg Lys Leu Pro Val Asn Arg Pro Ile 245 250 255 Asp Trp Lys Val Cys Gln Arg Ile Val Gly Leu Leu Gly Phe Ala Ala 260 265 270 Pro Phe Thr Gln Cys Gly Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys 275 280 285 Ile Gln Ser Lys Gln Ala Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe 290 295 300 Leu Cys Lys Gln Tyr Leu Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro 305 310 315 320 Gly Leu Cys Gln Val Phe Ala Ala Ala Thr Pro Thr Gly Trp Gly Leu 325 330 335 Val Met Gly His Gln Arg Met Arg Gly Thr Phe Leu Ala Pro Leu Pro 340 345 350 Ile His Thr Ala Ala Leu Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser 355 360 365 Gly Ala Asn Ile Leu Gly Thr Val Asn Ser Val Val Leu Ser Arg Lys 370 375 380 Tyr Thr Ser Phe Pro Trp Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu 385 390 395 400 Arg Gly Thr Ser Phe Val Tyr Val Pro Ser Ala Leu Asn Pro Ala Ala 405 410 415 Asp Pro Ser Arg Gly Arg Leu Gly Leu Ser Arg Pro Leu Leu Arg Leu 420 425 430 Pro Phe Arg Pro Thr Thr Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro 435 440 445 Ser Val Pro Ser His Leu Pro Asp Arg Val His Phe Ala Ser Pro Leu 450 455 460 His Val Ala Trp Arg Pro Pro 465 470 <210> 24 <211> 1410 <212> PRT <213> Artificial sequence <220> <223> Synthetic sequence HBV EpiC SPC FL <400> 24 Met Gly Gly Trp Ser Ser Lys Pro Arg Gln Gly Met Gly Thr Asn Leu 1 5 10 15 Ser Val Pro Asn Pro Leu Gly Phe Phe Pro Asp His Gln Leu Asp Pro 20 25 30 Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp Trp Asp Phe Asn Pro Asn 35 40 45 Lys Asp His Trp Pro Glu Ala Asn Gln Val Gly Ala Gly Ala Phe Gly 50 55 60 Pro Gly Phe Thr Pro Pro His Gly Gly Leu Leu Gly Trp Ser Pro Gln 65 70 75 80 Ala Gln Gly Ile Leu Thr Thr Val Pro Ala Ala Pro Pro Pro Ala Ser 85 90 95 Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr Pro Ile Ser Pro Pro Leu 100 105 110 Arg Asp Ser His Pro Gln Ala Gln Trp Asn Ser Thr Thr Phe His Gln 115 120 125 Ala Leu Leu Asp Pro Arg Val Arg Gly Leu Tyr Phe Pro Ala Gly Gly 130 135 140 Ser Ser Ser Gly Thr Val Asn Pro Val Pro Thr Thr Ala Ser Pro Ile 145 150 155 160 Ser Ser Ile Phe Ser Arg Thr Gly Asp Pro Ala Pro Asn Glu Asn Thr 165 170 175 Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe 180 185 190 Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp 195 200 205 Thr Ser Leu Asn Phe Leu Gly Gly Ala Pro Thr Cys Pro Gly Gln Asn 210 215 220 Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr Ser Cys Pro Pro Ile 225 230 235 240 Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe Ile Ile Phe Leu 245 250 255 Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr 260 265 270 Gln Gly Met Leu Pro Val Cys Pro Leu Leu Pro Gly Thr Ser Thr Thr 275 280 285 Ser Thr Gly Pro Cys Lys Thr Cys Thr Ile Pro Ala Gln Gly Thr Ser 290 295 300 Met Phe Pro Ser Cys Cys Cys Thr Lys Pro Ser Asp Gly Asn Cys Thr 305 310 315 320 Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Ala Arg Phe Leu Trp Glu 325 330 335 Trp Ala Ser Val Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val 340 345 350 Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp 355 360 365 Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Asn Ile Leu Ser Pro Phe 370 375 380 Leu Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp Val Tyr Ile Pro Leu 385 390 395 400 Ser Tyr Gln His Phe Arg Lys Leu Leu Leu Leu Asp Asp Glu Ala Gly 405 410 415 Pro Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp Glu Gly Leu Asn Arg 420 425 430 Arg Val Ala Glu Asp Leu Asn Leu Gly Asn Leu Asn Val Ser Ile Pro 435 440 445 Trp Thr His Lys Val Gly Asn Phe Thr Gly Leu Tyr Ser Ser Thr Val 450 455 460 Pro Val Phe Asn Pro Glu Trp Gln Thr Pro Ser Phe Pro His Ile His 465 470 475 480 Leu Gln Glu Asp Ile Ile Asn Arg Cys Gln Gln Tyr Val Gly Pro Leu 485 490 495 Thr Val Asn Glu Lys Arg Arg Leu Lys Leu Ile Met Pro Ala Arg Phe 500 505 510 Tyr Pro Asn Leu Thr Lys Tyr Leu Pro Leu Asp Lys Gly Ile Lys Pro 515 520 525 Tyr Tyr Pro Glu His Ala Val Asn His Tyr Phe Lys Thr Arg His Tyr 530 535 540 Leu His Thr Leu Trp Lys Ala Gly Ile Leu Tyr Lys Arg Glu Thr Thr 545 550 555 560 Arg Ser Ala Ser Phe Cys Gly Ser Pro Tyr Ser Trp Glu Gln Glu Leu 565 570 575 Gln His Gly Arg Leu Val Phe Gln Thr Ser Thr Arg His Gly Asp Glu 580 585 590 Ser Phe Cys Ser Gln Ser Ser Gly Ile Leu Ser Arg Ser Pro Val Gly 595 600 605 Pro Cys Val Arg Ser Gln Leu Lys Gln Ser Arg Leu Gly Leu Gln Pro 610 615 620 Gln Gln Gly Ser Leu Ala Arg Gly Lys Ser Gly Arg Ser Gly Ser Ile 625 630 635 640 Arg Ala Arg Val His Pro Thr Thr Arg Arg Ser Phe Gly Val Glu Pro 645 650 655 Ser Gly Ser Gly His Ile Asp Asn Ser Ala Ser Ser Thr Ser Ser Cys 660 665 670 Leu His Gln Ser Ala Val Arg Lys Thr Ala Tyr Ser His Leu Ser Thr 675 680 685 Ser Lys Arg Gln Ser Ser Ser Gly His Ala Val Glu Leu His Asn Ile 690 695 700 Pro Pro Ser Ser Ala Arg Ser Gln Ser Glu Gly Pro Ile Phe Ser Cys 705 710 715 720 Trp Trp Leu Gln Phe Arg Asn Ser Lys Pro Cys Ser Asp Tyr Cys Leu 725 730 735 Thr His Ile Val Asn Leu Leu Glu Asp Trp Gly Pro Cys Thr Glu His 740 745 750 Gly Glu His Asn Ile Arg Ile Pro Arg Thr Pro Ala Arg Val Thr Gly 755 760 765 Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr Thr Glu Ser Arg 770 775 780 Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Ser Thr His Val Ser 785 790 795 800 Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr Asn Leu Leu 805 810 815 Ser Ser Asn Leu Ser Trp Leu Ser Leu Asp Val Ser Ala Ala Phe Tyr 820 825 830 His Ile Pro Leu His Pro Ala Ala Met Pro His Leu Leu Val Gly Ser 835 840 845 Ser Gly Leu Pro Arg Tyr Val Ala Arg Leu Ser Ser Thr Ser Arg Asn 850 855 860 Ile Asn Tyr Gln His Gly Thr Met Gln Asp Leu His Asp Ser Cys Ser 865 870 875 880 Arg Asn Leu Tyr Val Ser Leu Leu Leu Leu Tyr Lys Thr Phe Gly Arg 885 890 895 Lys Leu His Leu Tyr Ser His Pro Ile Ile Leu Gly Phe Arg Lys Ile 900 905 910 Pro Met Gly Val Gly Leu Ser Pro Phe Leu Leu Ala Gln Phe Thr Ser 915 920 925 Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His Cys Leu Ala Phe 930 935 940 Ser Val Val Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe 945 950 955 960 Thr Ser Ile Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro 965 970 975 Asn Lys Thr Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val 980 985 990 Ile Gly Ser Trp Gly Thr Leu Pro Gln Glu His Ile Val Leu Lys Ile 995 1000 1005 Lys Gln Cys Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys 1010 1015 1020 Val Cys Gln Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr 1025 1030 1035 1040 Gln Cys Gly Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser 1045 1050 1055 Lys Gln Ala Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys 1060 1065 1070 Gln Tyr Leu Asn Leu Tyr Pro Val Ala Arg Gln Arg Ser Gly Leu Cys 1075 1080 1085 Gln Val Phe Ala Ala Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly 1090 1095 1100 His Arg Arg Met Arg Gly Thr Phe Val Ala Pro Leu Pro Ile His Thr 1105 1110 1115 1120 Ala Ala Leu Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys 1125 1130 1135 Leu Ile Gly Thr Val Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser 1140 1145 1150 Phe Pro Trp Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr 1155 1160 1165 Ser Phe Val Tyr Val Pro Ser Ala Leu Asn Pro Ala Ala Asp Pro Ser 1170 1175 1180 Arg Gly Arg Leu Gly Leu Tyr Arg Pro Leu Leu His Leu Pro Phe Arg 1185 1190 1195 1200 Pro Thr Thr Gly Arg Thr Ser Leu Tyr Ala Val Ser Pro Ser Val Pro 1205 1210 1215 Ser His Leu Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala 1220 1225 1230 Trp Arg Pro Pro Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val 1235 1240 1245 Glu Leu Leu Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp 1250 1255 1260 Leu Leu Asp Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro 1265 1270 1275 1280 Glu His Cys Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys 1285 1290 1295 Trp Gly Glu Leu Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu 1300 1305 1310 Asp Pro Ala Ser Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met 1315 1320 1325 Gly Leu Lys Ile Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr 1330 1335 1340 Phe Gly Arg Glu Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp 1345 1350 1355 1360 Ile Arg Thr Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser 1365 1370 1375 Thr Leu Pro Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg 1380 1385 1390 Arg Arg Thr Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Arg Glu Ser 1395 1400 1405 Gln Cys 1410 <210> 25 <211> 1183 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence HBV EpiC SPC Δpol612-838 <400> 25 Met Gly Gly Trp Ser Ser Lys Pro Arg Gln Gly Met Gly Thr Asn Leu 1 5 10 15 Ser Val Pro Asn Pro Leu Gly Phe Phe Pro Asp His Gln Leu Asp Pro 20 25 30 Ala Phe Gly Ala Asn Ser Asn Asn Pro Asp Trp Asp Phe Asn Pro Asn 35 40 45 Lys Asp His Trp Pro Glu Ala Asn Gln Val Gly Ala Gly Ala Phe Gly 50 55 60 Pro Gly Phe Thr Pro Pro His Gly Gly Leu Leu Gly Trp Ser Pro Gln 65 70 75 80 Ala Gln Gly Ile Leu Thr Thr Val Pro Ala Ala Pro Pro Pro Ala Ser 85 90 95 Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr Pro Ile Ser Pro Pro Leu 100 105 110 Arg Asp Ser His Pro Gln Ala Gln Trp Asn Ser Thr Thr Phe His Gln 115 120 125 Ala Leu Leu Asp Pro Arg Val Arg Gly Leu Tyr Phe Pro Ala Gly Gly 130 135 140 Ser Ser Ser Gly Thr Val Asn Pro Val Pro Thr Thr Ala Ser Pro Ile 145 150 155 160 Ser Ser Ile Phe Ser Arg Thr Gly Asp Pro Ala Pro Asn Glu Asn Thr 165 170 175 Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu Gln Ala Gly Phe Phe 180 185 190 Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser Leu Asp Ser Trp Trp 195 200 205 Thr Ser Leu Asn Phe Leu Gly Gly Ala Pro Thr Cys Pro Gly Gln Asn 210 215 220 Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr Ser Cys Pro Pro Ile 225 230 235 240 Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe Ile Ile Phe Leu 245 250 255 Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr 260 265 270 Gln Gly Met Leu Pro Val Cys Pro Leu Leu Pro Gly Thr Ser Thr Thr 275 280 285 Ser Thr Gly Pro Cys Lys Thr Cys Thr Ile Pro Ala Gln Gly Thr Ser 290 295 300 Met Phe Pro Ser Cys Cys Cys Thr Lys Pro Ser Asp Gly Asn Cys Thr 305 310 315 320 Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Ala Arg Phe Leu Trp Glu 325 330 335 Trp Ala Ser Val Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val 340 345 350 Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp 355 360 365 Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Asn Ile Leu Ser Pro Phe 370 375 380 Leu Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp Val Tyr Ile Pro Leu 385 390 395 400 Ser Tyr Gln His Phe Arg Lys Leu Leu Leu Leu Asp Asp Glu Ala Gly 405 410 415 Pro Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp Glu Gly Leu Asn Arg 420 425 430 Arg Val Ala Glu Asp Leu Asn Leu Gly Asn Leu Asn Val Ser Ile Pro 435 440 445 Trp Thr His Lys Val Gly Asn Phe Thr Gly Leu Tyr Ser Ser Thr Val 450 455 460 Pro Val Phe Asn Pro Glu Trp Gln Thr Pro Ser Phe Pro His Ile His 465 470 475 480 Leu Gln Glu Asp Ile Ile Asn Arg Cys Gln Gln Tyr Val Gly Pro Leu 485 490 495 Thr Val Asn Glu Lys Arg Arg Leu Lys Leu Ile Met Pro Ala Arg Phe 500 505 510 Tyr Pro Asn Leu Thr Lys Tyr Leu Pro Leu Asp Lys Gly Ile Lys Pro 515 520 525 Tyr Tyr Pro Glu His Ala Val Asn His Tyr Phe Lys Thr Arg His Tyr 530 535 540 Leu His Thr Leu Trp Lys Ala Gly Ile Leu Tyr Lys Arg Glu Thr Thr 545 550 555 560 Arg Ser Ala Ser Phe Cys Gly Ser Pro Tyr Ser Trp Glu Gln Glu Leu 565 570 575 Gln His Gly Arg Leu Val Phe Gln Thr Ser Thr Arg His Gly Asp Glu 580 585 590 Ser Phe Cys Ser Gln Ser Ser Gly Ile Leu Ser Arg Ser Pro Val Gly 595 600 605 Pro Cys Val Arg Ser Gln Leu Lys Gln Ser Arg Leu Gly Leu Gln Pro 610 615 620 Gln Gln Gly Ser Leu Ala Arg Gly Lys Ser Gly Arg Ser Gly Ser Ile 625 630 635 640 Arg Ala Arg Val His Pro Thr Thr Arg Arg Ser Phe Gly Val Glu Pro 645 650 655 Ser Gly Ser Gly His Ile Asp Asn Ser Ala Ser Ser Thr Ser Ser Cys 660 665 670 Leu His Gln Ser Ala Val Arg Lys Thr Ala Tyr Ser His Leu Ser Thr 675 680 685 Ser Lys Arg Gln Ser Ser Ser Gly His Ala Val Glu Leu His Asn Ile 690 695 700 Pro Pro Ser Ser Ala Arg Ser Gln Ser Glu Gly Pro Ile Phe Ser Cys 705 710 715 720 Trp Trp Leu Gln Phe Arg Asn Ser Lys Pro Cys Ser Asp Tyr Cys Leu 725 730 735 Thr His Ile Val Asn Leu Leu Glu Asp Trp Gly Pro Cys Thr Glu His 740 745 750 Gly Glu His Asn Ile Arg Ile Pro Arg Thr Pro Ala Arg Val Thr Gly 755 760 765 Gly Val Phe Leu Val Asp Lys Asn Pro His Asn Thr Thr Glu Ser Arg 770 775 780 Leu Val Val Asp Phe Ser Gln Phe Ser Arg Gly Ser Thr His Val Ser 785 790 795 800 Trp Pro Lys Phe Ala Val Pro Asn Leu Gln Ser Leu Thr Asn Leu Leu 805 810 815 Arg Ser Asn Leu Ser Trp Leu Ser Leu Asp Val Ser Ala Ala Phe Tyr 820 825 830 His Ile Pro Leu His Pro Ala Ala Met Pro His Leu Leu Val Gly Ser 835 840 845 Ser Gly Leu Pro Arg Tyr Val Ala Arg Leu Ser Ser Thr Ser Arg Asn 850 855 860 Ile Asn Tyr Gln His Gly Thr Met Gln Asp Leu His Asp Ser Cys Ser 865 870 875 880 Arg Asn Leu Tyr Val Ser Leu Leu Leu Leu Tyr Lys Thr Phe Gly Arg 885 890 895 Lys Leu His Leu Tyr Ser His Pro Ile Ile Leu Gly Phe Arg Lys Ile 900 905 910 Pro Met Gly Val Gly Leu Ser Pro Phe Leu Leu Ala Gln Phe Thr Ser 915 920 925 Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His Cys Leu Ala Phe 930 935 940 Ser Val Val Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Phe 945 950 955 960 Thr Ser Ile Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro 965 970 975 Asn Lys Thr Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val 980 985 990 Ile Gly Ser Trp Gly Thr Leu Pro Gln Glu His Ile Val Leu Lys Ile 995 1000 1005 Lys Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu 1010 1015 1020 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Ile Arg Asp Leu Leu Asp 1025 1030 1035 1040 Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys 1045 1050 1055 Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 1060 1065 1070 Leu Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala 1075 1080 1085 Ser Arg Glu Leu Val Val Ser Tyr Val Asn Val Asn Met Gly Leu Lys 1090 1095 1100 Ile Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg 1105 1110 1115 1120 Glu Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr 1125 1130 1135 Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro 1140 1145 1150 Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr 1155 1160 1165 Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Arg Glu Ser Gln Cys 1170 1175 1180 <210> 26 <211> 1169 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence HBV EpiD SPC Δpol601 - 827 <400> 26 Met Gly Gln Asn Leu Ser Thr Ser Asn Pro Leu Gly Phe Phe Pro Asp 1 5 10 15 His Gln Leu Asp Pro Ala Phe Arg Ala Asn Thr Ala Asn Pro Asp Trp 20 25 30 Asp Phe Asn Pro Asn Lys Asp Thr Trp Pro Asp Ala Asn Lys Val Gly 35 40 45 Ala Gly Ala Phe Gly Leu Gly Phe Thr Pro Pro His Gly Gly Leu Leu 50 55 60 Gly Trp Ser Pro Gln Ala Gln Gly Ile Leu Gln Thr Leu Pro Ala Asn 65 70 75 80 Pro Pro Pro Ala Ser Thr Asn Arg Gln Ser Gly Arg Gln Pro Thr Pro 85 90 95 Leu Ser Pro Pro Leu Arg Asn Thr His Pro Gln Ala Gln Trp Asn Ser 100 105 110 Thr Thr Phe His Gln Thr Leu Gln Asp Pro Arg Val Arg Gly Leu Tyr 115 120 125 Phe Pro Ala Gly Gly Ser Ser Ser Gly Thr Val Asn Pro Val Pro Thr 130 135 140 Thr Val Ser His Ile Ser Ser Ile Phe Ser Arg Ile Gly Asp Pro Ala 145 150 155 160 Leu Asn Glu Asn Ile Thr Ser Gly Phe Leu Gly Pro Leu Leu Val Leu 165 170 175 Gln Ala Gly Phe Phe Leu Leu Thr Arg Ile Leu Thr Ile Pro Gln Ser 180 185 190 Leu Asp Ser Trp Trp Thr Ser Leu Asn Phe Leu Gly Gly Thr Thr Val 195 200 205 Cys Leu Gly Gln Asn Ser Gln Ser Pro Thr Ser Asn His Ser Pro Thr 210 215 220 Ser Cys Pro Pro Thr Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg 225 230 235 240 Phe Ile Ile Phe Leu Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu 245 250 255 Val Leu Leu Asp Tyr Gln Gly Met Leu Pro Val Cys Pro Leu Ile Pro 260 265 270 Gly Ser Ser Thr Thr Ser Thr Gly Pro Cys Arg Thr Cys Thr Thr Pro 275 280 285 Ala Gln Gly Thr Ser Met Tyr Pro Ser Cys Cys Cys Thr Lys Pro Ser 290 295 300 Asp Gly Asn Cys Thr Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Gly 305 310 315 320 Lys Phe Leu Trp Glu Trp Ala Ser Ala Arg Phe Ser Trp Leu Ser Leu 325 330 335 Leu Val Pro Phe Val Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp 340 345 350 Leu Ser Val Ile Trp Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Ser 355 360 365 Ile Leu Ser Pro Phe Leu Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp 370 375 380 Val Tyr Ile Pro Leu Ser Tyr Gln His Phe Arg Arg Leu Leu Leu Leu 385 390 395 400 Asp Asp Glu Ala Gly Pro Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp 405 410 415 Glu Gly Leu Asn Arg Arg Val Ala Glu Asp Leu Asn Leu Gly Asn Leu 420 425 430 Asn Val Ser Ile Pro Trp Thr His Lys Val Gly Asn Phe Thr Gly Leu 435 440 445 Tyr Ser Ser Thr Val Pro Val Phe Asn Pro His Trp Lys Thr Pro Ser 450 455 460 Phe Pro Asn Ile His Leu His Gln Asp Ile Ile Lys Lys Cys Glu Gln 465 470 475 480 Phe Val Gly Pro Leu Thr Val Asn Glu Lys Arg Arg Leu Gln Leu Ile 485 490 495 Met Pro Ala Arg Phe Tyr Pro Asn Val Thr Lys Tyr Leu Pro Leu Asp 500 505 510 Lys Gly Ile Lys Pro Tyr Tyr Pro Glu His Leu Val Asn His Tyr Phe 515 520 525 Gln Thr Arg His Tyr Leu His Thr Leu Trp Lys Ala Gly Ile Leu Tyr 530 535 540 Lys Arg Glu Thr Thr His Ser Ala Ser Phe Cys Gly Ser Pro Tyr Ser 545 550 555 560 Trp Glu Gln Glu Leu Gln His Gly Ala Glu Ser Phe His Gln Gln Ser 565 570 575 Ser Gly Ile Leu Ser Arg Pro Pro Val Gly Ser Ser Leu Gln Ser Lys 580 585 590 His Arg Lys Ser Arg Leu Gly Leu Gln Ser Gln Gln Gly His Leu Ala 595 600 605 Arg Arg Gln Gln Gly Arg Ser Trp Ser Ile Arg Ala Gly Ile His Pro 610 615 620 Thr Ala Arg Arg Pro Phe Gly Val Glu Pro Ser Gly Ser Gly His Thr 625 630 635 640 Thr Asn Leu Ala Ser Lys Ser Ala Ser Cys Leu Tyr Gln Ser Pro Val 645 650 655 Arg Lys Ala Ala Tyr Pro Ala Val Ser Thr Phe Glu Lys His Ser Ser 660 665 670 Ser Gly His Ala Val Glu Leu His Asn Leu Pro Pro Asn Ser Ala Arg 675 680 685 Ser Gln Ser Glu Arg Pro Val Phe Pro Cys Trp Trp Leu Gln Phe Arg 690 695 700 Asn Ser Lys Pro Cys Ser Asp Tyr Cys Leu Ser His Ile Val Asn Leu 705 710 715 720 Leu Glu Asp Trp Gly Pro Cys Ala Glu His Gly Glu His His Ile Arg 725 730 735 Ile Pro Arg Thr Pro Ala Arg Val Thr Gly Gly Val Phe Leu Val Asp 740 745 750 Lys Asn Pro His Asn Thr Ala Glu Ser Arg Leu Val Val Asp Phe Ser 755 760 765 Gln Phe Ser Arg Gly Asn Tyr Arg Val Ser Trp Pro Lys Phe Ala Val 770 775 780 Pro Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp 785 790 795 800 Leu Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro 805 810 815 Ala Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr 820 825 830 Val Ala Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn His Gln His Gly 835 840 845 Thr Met Gln Asn Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser 850 855 860 Leu Leu Leu Leu Tyr Gln Thr Phe Gly Arg Lys Leu His Leu Tyr Ser 865 870 875 880 His Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu 885 890 895 Ser Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val 900 905 910 Arg Arg Ala Phe Pro His Cys Leu Ala Phe Ser Val Val Leu Gly Ala 915 920 925 Lys Ser Val Gln His Leu Glu Ser Leu Phe Thr Ala Val Thr Asn Phe 930 935 940 Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr Lys Arg Trp 945 950 955 960 Gly Tyr Ser Leu His Phe Met Gly Tyr Val Ile Gly Cys Tyr Gly Ser 965 970 975 Leu Pro Gln Asp His Ile Ile Gln Lys Ile Lys Asp Ile Asp Pro Tyr 980 985 990 Lys Glu Phe Gly Ala Thr Val Glu Leu Leu Ser Phe Leu Pro Ser Asp 995 1000 1005 Phe Phe Pro Ser Val Arg Asp Leu Leu Asp Thr Ala Ser Ala Leu Tyr 1010 1015 1020 Arg Glu Ala Leu Glu Ser Pro Glu His Cys Ser Pro His His Thr Ala 1025 1030 1035 1040 Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu Leu Met Thr Leu Ala Thr 1045 1050 1055 Trp Val Gly Gly Asn Leu Glu Asp Pro Ile Ser Arg Asp Leu Val Val 1060 1065 1070 Ser Tyr Val Asn Thr Asn Met Gly Leu Lys Phe Arg Gln Leu Leu Trp 1075 1080 1085 Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu Thr Val Ile Glu Tyr 1090 1095 1100 Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro Pro Ala Tyr Arg Pro 1105 1110 1115 1120 Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu Thr Thr Val Val Arg 1125 1130 1135 Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro Ser Pro Arg Arg Arg 1140 1145 1150 Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser Lys Ser Arg Glu Ser Gln 1155 1160 1165 Cys <210> 27 <211> 67 <212> PRT <213> artificial sequence <220> <223> synthetic sequence 1epi_C1 <400> 27 Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu 1 5 10 15 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp 20 25 30 Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys 35 40 45 Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 50 55 60 Leo Met Asn 65 <210> 28 <211> 79 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 1epi_C2 <400> 28 Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg Glu 1 5 10 15 Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr Pro 20 25 30 Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro Glu 35 40 45 Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr Pro 50 55 60 Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser 65 70 75 <210> 29 <211> 158 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 1epi_S1 <400> 29 Cys Pro Gly Tyr Arg Trp Met Cys Leu Arg Arg Phe Ile Ile Phe Leu 1 5 10 15 Phe Ile Leu Leu Leu Cys Leu Ile Phe Leu Leu Val Leu Leu Asp Tyr 20 25 30 Gln Gly Met Leu Pro Val Cys Pro Leu Ile Pro Gly Ser Ser Thr Thr 35 40 45 Ser Thr Gly Pro Cys Lys Thr Cys Thr Thr Pro Ala Gln Gly Thr Ser 50 55 60 Met Phe Pro Ser Cys Cys Cys Thr Lys Pro Ser Asp Gly Asn Cys Thr 65 70 75 80 Cys Ile Pro Ile Pro Ser Ser Trp Ala Phe Ala Lys Tyr Leu Trp Glu 85 90 95 Trp Ala Ser Val Arg Phe Ser Trp Leu Ser Leu Leu Val Pro Phe Val 100 105 110 Gln Trp Phe Val Gly Leu Ser Pro Thr Val Trp Leu Ser Val Ile Trp 115 120 125 Met Met Trp Tyr Trp Gly Pro Ser Leu Tyr Asn Ile Leu Ser Pro Phe 130 135 140 Leu Pro Leu Leu Pro Ile Phe Phe Cys Leu Trp Val Tyr Ile 145 150 155 <210> 30 <211> 68 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 1epi_P1 <400> 30 Met Pro Leu Ser Tyr Gln His Phe Arg Lys Leu Leu Leu Leu Asp Asp 1 5 10 15 Glu Ala Gly Pro Leu Glu Glu Glu Leu Pro Arg Leu Ala Asp Glu Gly 20 25 30 Leu Asn Arg Arg Val Ala Glu Asp Leu Asn Leu Gly Asn Leu Asn Val 35 40 45 Ser Ile Pro Trp Thr His Lys Val Gly Asn Phe Thr Gly Leu Tyr Ser 50 55 60 Thr Val Pro Serum 65 <210> 31 <211> 104 <212> PRT <213> artificial sequence <220> <223> Synthetic sequence 1epi_P2a <400> 31 Pro Arg Thr Pro Ala Arg Val Thr Gly Gly Val Phe Leu Val Asp Lys 1 5 10 15 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 20 25 30 Phe Ser Arg Gly Asn Thr Arg Val Ser Trp Pro Lys Phe Ala Val Pro 35 40 45 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 50 55 60 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala 65 70 75 80 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val 85 90 95 Ala Arg Leu Ser Ser Asn Ser Arg 100 <210> 32 <211> 69 <212> PRT <213> Artificial sequence <220> <223> Synthetic sequence 1epiP2b <400> 32 Gly Arg Lys Leu His Leu Tyr Ser His Pro Ile Ile Leu Gly Phe Arg 1 5 10 15 Lys Ile Pro Met Gly Val Gly Leu Ser Pro Phe Leu Leu Ala Gln Phe 20 25 30 Thr Ser Ala Ile Cys Ser Val Val Arg Arg Ala Phe Pro His Cys Leu 35 40 45 Ala Phe Ser Tyr Met Asp Asp Val Val Leu Gly Ala Lys Ser Val Gln 50 55 60 His Leu Glu Ser Leu 65 <210> 33 <211> 226 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence 1epi_P2c <400> 33 Cys Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys 1 5 10 15 Gln Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys 20 25 30 Gly Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln 35 40 45 Ala Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr 50 55 60 Leu Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val 65 70 75 80 Phe Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Gln 85 90 95 Arg Met Arg Gly Thr Phe Val Ala Pro Leu Pro Ile His Thr Ala Glu 100 105 110 Leu Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile 115 120 125 Gly Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro 130 135 140 Trp Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe 145 150 155 160 Val Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly 165 170 175 Arg Leu Gly Leu Tyr Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr 180 185 190 Thr Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His 195 200 205 Leu Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg 210 215 220 Pro Pro 225 <210> 34 <211> 176 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 1epi_C <400> 34 Met Asp Ile Asp Pro Tyr Lys Glu Phe Gly Ala Ser Val Glu Leu Leu 1 5 10 15 Ser Phe Leu Pro Ser Asp Phe Phe Pro Ser Val Arg Asp Leu Leu Asp 20 25 30 Thr Ala Ser Ala Leu Tyr Arg Glu Ala Leu Glu Ser Pro Glu His Cys 35 40 45 Ser Pro His His Thr Ala Leu Arg Gln Ala Ile Leu Cys Trp Gly Glu 50 55 60 Leu Met Asn Leu Ala Thr Trp Val Gly Ser Asn Leu Glu Asp Pro Ala 65 70 75 80 Ser Arg Asp Leu Val Val Ser Tyr Val Asn Thr Asn Met Gly Leu Lys 85 90 95 Ile Arg Gln Leu Leu Trp Phe His Ile Ser Cys Leu Thr Phe Gly Arg 100 105 110 Glu Thr Val Leu Glu Tyr Leu Val Ser Phe Gly Val Trp Ile Arg Thr 115 120 125 Pro Pro Ala Tyr Arg Pro Pro Asn Ala Pro Ile Leu Ser Thr Leu Pro 130 135 140 Glu Thr Thr Val Val Arg Arg Arg Gly Arg Ser Pro Arg Arg Arg Thr 145 150 155 160 Pro Ser Pro Arg Arg Arg Arg Ser Gln Ser Pro Arg Arg Arg Arg Ser 165 170 175 <210> 35 <211> 481 <212> PRT <213> artificial sequence <220> <223> Synthesis sequence 1epi_P2 <400> 35 Pro Arg Thr Pro Ala Arg Val Thr Gly Gly Val Phe Leu Val Asp Lys 1 5 10 15 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 20 25 30 Phe Ser Arg Gly Asn Thr Arg Val Ser Trp Pro Lys Phe Ala Val Pro 35 40 45 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 50 55 60 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala 65 70 75 80 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val 85 90 95 Ala Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn His Gln His Gly Thr 100 105 110 Met Gln Asn Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 115 120 125 Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 130 135 140 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 145 150 155 160 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 165 170 175 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 180 185 190 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Tyr Thr Ala Val 195 200 205 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 210 215 220 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 225 230 235 240 Trp Gly Thr Leu Pro Gln Glu His Ile Val Gln Lys Ile Lys Met Cys 245 250 255 Phe Arg Lys Leu Pro Val Asn Arg Pro Ile Asp Trp Lys Val Cys Gln 260 265 270 Arg Ile Val Gly Leu Leu Gly Phe Ala Ala Pro Phe Thr Gln Cys Gly 275 280 285 Tyr Pro Ala Leu Met Pro Leu Tyr Ala Cys Ile Gln Ser Lys Gln Ala 290 295 300 Phe Thr Phe Ser Pro Thr Tyr Lys Ala Phe Leu Cys Lys Gln Tyr Leu 305 310 315 320 Asn Leu Tyr Pro Val Ala Arg Gln Arg Pro Gly Leu Cys Gln Val Phe 325 330 335 Ala Asp Ala Thr Pro Thr Gly Trp Gly Leu Ala Ile Gly His Gln Arg 340 345 350 Met Arg Gly Thr Phe Val Ala Pro Leu Pro Ile His Thr Ala Glu Leu 355 360 365 Leu Ala Ala Cys Phe Ala Arg Ser Arg Ser Gly Ala Lys Leu Ile Gly 370 375 380 Thr Asp Asn Ser Val Val Leu Ser Arg Lys Tyr Thr Ser Phe Pro Trp 385 390 395 400 Leu Leu Gly Cys Ala Ala Asn Trp Ile Leu Arg Gly Thr Ser Phe Val 405 410 415 Tyr Val Pro Ser Ala Leu Asn Pro Ala Asp Asp Pro Ser Arg Gly Arg 420 425 430 Leu Gly Leu Tyr Arg Pro Leu Leu Arg Leu Pro Phe Arg Pro Thr Thr 435 440 445 Gly Arg Thr Ser Leu Tyr Ala Asp Ser Pro Ser Val Pro Ser His Leu 450 455 460 Pro Asp Arg Val His Phe Ala Ser Pro Leu His Val Ala Trp Arg Pro 465 470 475 480 Pro <210> 36 <211> 254 <212> PRT <213> Artificial Sequence <220> <223> Synthetic Sequence 1epi_P2 minus P2c <400> 36 Pro Arg Thr Pro Ala Arg Val Thr Gly Gly Val Phe Leu Val Asp Lys 1 5 10 15 Asn Pro His Asn Thr Thr Glu Ser Arg Leu Val Val Asp Phe Ser Gln 20 25 30 Phe Ser Arg Gly Asn Thr Arg Val Ser Trp Pro Lys Phe Ala Val Pro 35 40 45 Asn Leu Gln Ser Leu Thr Asn Leu Leu Ser Ser Asn Leu Ser Trp Leu 50 55 60 Ser Leu Asp Val Ser Ala Ala Phe Tyr His Leu Pro Leu His Pro Ala 65 70 75 80 Ala Met Pro His Leu Leu Val Gly Ser Ser Gly Leu Ser Arg Tyr Val 85 90 95 Ala Arg Leu Ser Ser Asn Ser Arg Ile Phe Asn His Gln His Gly Thr 100 105 110 Met Gln Asn Leu His Asp Ser Cys Ser Arg Asn Leu Tyr Val Ser Leu 115 120 125 Leu Leu Leu Tyr Lys Thr Phe Gly Arg Lys Leu His Leu Tyr Ser His 130 135 140 Pro Ile Ile Leu Gly Phe Arg Lys Ile Pro Met Gly Val Gly Leu Ser 145 150 155 160 Pro Phe Leu Leu Ala Gln Phe Thr Ser Ala Ile Cys Ser Val Val Arg 165 170 175 Arg Ala Phe Pro His Cys Leu Ala Phe Ser Tyr Met Asp Asp Val Val 180 185 190 Leu Gly Ala Lys Ser Val Gln His Leu Glu Ser Leu Tyr Thr Ala Val 195 200 205 Thr Asn Phe Leu Leu Ser Leu Gly Ile His Leu Asn Pro Asn Lys Thr 210 215 220 Lys Arg Trp Gly Tyr Ser Leu Asn Phe Met Gly Tyr Val Ile Gly Ser 225 230 235 240 Trp Gly Thr Leu Pro Gln Glu His Ile Val Gln Lys Ile Lys 245 250

Claims

1. A polypeptide consisting of the amino acid sequence shown in SEQ ID NO:

15.

2. A cytomegalovirus (CMV) vector comprising a polynucleotide encoding the polypeptide of SEQ ID NO:

15.

3. The CMV vector of claim 2, wherein the CMV vector is a human CMV (HCMV) vector or a rhesus CMV (RhCMV) vector.

4. The CMV vector of claim 3, wherein the CMV vector lacks the UL82 gene.

5. The CMV vector of claim 3, wherein the CMV vector lacks the UL128-UL130 gene region and lacks the UL146-UL147 gene region.

6. The CMV vector of claim 3, wherein the CMV vector has an intact UL128-UL130 gene region and an intact UL146-UL147 gene region.

7. A composition comprising two or more CMV vectors according to claim 3.

8. A hepatitis B virus (HBV) vaccine comprising the polypeptide according to claim 1 and a pharmaceutically acceptable carrier.

9. A hepatitis B virus (HBV) vaccine comprising the CMV vector according to any one of claims 2 to 6 and a pharmaceutically acceptable carrier.

10. Use of the polypeptide according to claim 1 in the preparation of a medicament for treating HBV.

11. Use of a carrier according to any one of claims 2 to 6 in the manufacture of a medicament for treating HBV.

12. Use of the vaccine according to claim 9 in the manufacture of a medicament for the treatment of HBV.

13. Use of the polypeptide according to claim 1 in the preparation of a medicament for protecting a subject from HBV infection.

14. Use of a carrier according to any one of claims 2 to 6 in the manufacture of a medicament for protecting a subject from HBV infection.

15. Use of a vaccine according to claim 9 in the manufacture of a medicament for protecting a subject from HBV infection.

16. Use of the polypeptide according to claim 1 in the preparation of a medicament for generating or inducing an immune response to HBV in a subject.

17. Use of a vector according to any one of claims 2 to 6 in the manufacture of a medicament for generating or inducing an immune response to HBV in a subject.

18. Use of a vaccine according to claim 8 or 9 in the manufacture of a medicament for generating or inducing an immune response to HBV in a subject.

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