DESC:Kyasanur Forest Disease Virus envelope protein peptide based vaccine and method thereof

Field of Invention:
The present invention relates to a field of Kyasanur Forest Disease Virus envelope protein peptide based vaccine and method thereof.
Background of the invention:
The following background discussion includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art.

Traditional viral vaccines derived from whole virus or virus-derived are successfully employed globally. Unfortunately, a large number of infectious diseases resulting from flaviviruses infection do not have efficacious vaccines. A key hindrance in advocating whole-pathogen or pathogen-derived subunit strategy in flavivirus is due to unwanted ADE in the vaccinated host that may fail to protect or even cause harm. Hence, the essential ingredients for developing effective vaccines against flaviviruses are targeted immunogen with precision for developing mAb-like neutralizing antibodies while limiting ADE.
Kyasanur Forest Disease (KFD) is caused by Kyasanur Forest Disease Virus (KFDV) first time reported in 1957 people living near the Kyasanur forest in the Shimoga district of Karnataka state, India. Later on, it is extended to other parts of India, such a Kerala, Tamil Nadu, Goa, and Gujarat. The fatality rate for the disease is 3-10%. KFDV is a member of the tick-borne human pathogenic flavivirus. Despite an estimated annual incidence of 400-500 cases in India, KFD is historically understudied. There are no proven antivirals, or FDA approved vaccines against KFDV. These viruses have a positive-sense, single-standard RNA genome packaged in a viral envelope. The viral envelope is highly immunogenic and is a target for neutralizing antibody response. Hence, polyclonal antibodies developed on natural infection of a flavivirus (KFDV) could cross-react to another flavivirus such as tick-borne encephalitis virus, TBEV. Paradoxically, this cross-reactivity of antibodies may enhance the new virus infection and could exacerbate the disease condition. This “antibody-mediated-enhancement (ADE)” effect is a key challenge is vaccine designing. Hence the gold-standard vaccine candidate must induce a protective neutralizing antibody response devoid of any cross-reactive antibodies in the host.
Currently, the formalin-inactivated KFDV vaccine is being used. However, its efficacy is low, and need to be given in multiple buster doses. The known KFDV vaccine is a formalin Inactivated virus. Due to its strict biosafety (BSL-4) and a very few labs get permission to work with this virus. Due to this, there are no effective vaccines against KFDV.

Hence, there is a need of an immunogenic peptides and discontinuous epitopes as vaccine candidates against KFDV.

Object of the invention:
Primary object of the present invention is to overcome the drawback associated with the prior art.
Another object of the present invention is to provide vaccine candidate against KFDV.

Another object of the present invention is to provide an immunogenic peptides and discontinuous epitopes as vaccine candidates against KFDV.

Another object of the present invention is to provide dominant immunizing envelope protein peptides as vaccine candidates against KFDV.

Another object of the present invention is to provide vaccine comprising immunodominant peptides.

Summary of the Invention:
The Invention provides a Kyasanur Forest Disease Virus envelope protein peptide based vaccine comprising:
a) 15 T-cell comprising MHC-I and MHC-II, dominant epitopes;
b) 11 linear and seven discontinuous B-cell epitopes

In an embodiment, the KFDV MHC-I dominant epitopes comprises:
KFDV MHC-I
Epitopes HLA alleles Start -End
TRVSLVLEL HLA-B*39:01 19-27
LTAEGKPSV HLA-A*69:01 33-41
KTREYCLHAKL HLA-A*30:01,HLA-B*40:01,HLA-B*40:02 55-65
CPAMGPATL HLA-B*07:02,HLA-B*35:01,HLA-B*07:02 74-82
KKKATGYVY HLA-B*15:03 124-132
HSNRKTASF HLA-B*15:03,HLA-B*15:17,HLA-B*58:01 157-165
LPWRHGGAQ HLA-B*07:02 225-233
QEWNHADRL HLA-B*40:01 233-241
METTGGGFV HLA-B*40:01 364-372
YVGELSHQW HLA-B*58:01 384-392
SVGGMLSSV HLA-A*02:03 425-433
ALHTAFGAAF HLA-A*02:03,HLA-B*15:03 436-445
LVLTMTLGV HLA-A*02:01,HLA-A*02:06 486-494

In an embodiment, the KFDV MHC-II dominant epitopes comprises:
KFDV MHC-II
Epitopes HLA alleles Start -End
VRAVAHGEPNVNVAS HLA-DQA1*05:05/DQB1*03:13, HLA-DQA1*05:04/DQB1*03:13, HLA-DQA1*05:03/DQB1*03:13, HLA-DQA1*05:01/DQB1*03:13, HLA-DQA1*02:01/DQB1*03:13, HLA-DQA1*05:06/DQB1*03:13, HLA-DQA1*05:07/DQB1*03:13, HLA-DQA1*05:08/DQB1*03:13, HLA-DQA1*05:09/DQB1*03:13, HLA-DQA1*05:10/DQB1*03:13, HLA-DQA1*05:11/DQB1*03:13 342-356
VGGMLSSVGKALHTA HLA-DRB1*14:74, HLA-DRB1*09:06 426-440

In an embodiment, the KFDV B-cell linear epitopes comprises:
Epitopes KFDV
Epitope-1 5HLQNR9
Epitope-2 48IHQENPAKT56
Epitope-3 68SKVAARCPAMGPATLPEEHQASTVCRRDQSDRGWGN103
Epitope-4 148GDYLAANESHSNR160
Epitope-5 191GVD193
Epitope-6 204KTAEHLPKA212
Epitope-7 218DWFEDLSLPWRHGGAQEWNHAD239
Epitope-8 311KFAWKRPPTDSG322
Epitope-9 348GEPN351
Epitope-10 391HQWFQKGST398
Epitope-11 420AWDFGSVG427

Detailed Description of the drawing:
To further clarify advantages and features of the present invention, a more particular description of the invention will be rendered by reference to specific embodiments thereof, which is illustrated in the appended drawings. It is appreciated that these drawings depict only typical embodiments of the invention and are therefore not to be considered limiting of its scope. The invention will be described and explained with additional specificity and detail with the accompanying drawings in which:
Figure 1: illustrates KFDV Envelope protein modeled dimer and trimer.
Figure 2: illustrates KFDV envelope protein conformational epitopes
Detailed description of the Invention:
For the purpose of promoting an understanding of the principles of the invention, reference will now be made to the embodiment illustrated in the drawings and specific language will be used to describe the same. It will nevertheless be understood that no limitation of the scope of the invention is thereby intended, such alterations and further modifications in the illustrated system, and such further applications of the principles of the invention as illustrated therein being contemplated as would normally occur to one skilled in the art to which the invention relates.
It will be understood by those skilled in the art that the foregoing general description and the following detailed description are exemplary and explanatory of the invention and are not intended to be restrictive thereof.
Reference throughout this specification to “an embodiment, “another embodiment, “an or similar language means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment of the present invention. Thus, appearances of the phrase “in an embodiment, “in another embodiment, “in one and similar language throughout this specification may, but do not necessarily, all refer to the same embodiment.
The present invention relates to the field of Kyasanur Forest Disease Virus envelope protein peptide based vaccine and method thereof.
The invention provides immunizing a peptide mixture as a vaccine against tick-borne flavivirus infection, namely Kyasanur Forest Disease Virus (KFDV. The formulation has a mixture of 10 to 30 amino acids (aa)-long peptides derived from the amino acid sequence of a viral envelope protein of KFDV virus. The peptide sequences are derived from the results of computational biology and immuno-informatics analysis. Based on this knowledge, these peptides could activate specific T-cells through the host MHC/HLA genotype that would recognize these peptides to be processed by professional antigen-presenting cells (APCs). Further, these peptides will preferentially generate neutralizing antibodies in the host. While developing the peptides, specific customization was performed for reducing or eliminating antibody-mediated enhancement effect, an unwanted immune response associated with any Flavivirus infection.
In an embodiment, the vaccine comprising peptide mixture is used for developing recombinant monoclonal antibodies.
In an embodiment, the vaccine acts as an immunogen that induces a protective immune response against KFDV infection.

In an embodiment, immune-dominant epitopes present across the said viruses, were identified by comparing various parameters, that will produce neutralizing antibodies and not cross-reactive antibodies.

In an embodiment, the invention provides array of epitope-based immunogens capable of inducing immune response towards critical neutralizing antibodies. Simultaneously, avoiding the elicitation of cross-reactive antibodies that play no role in the protection and may even be harmful. This peptide-epitope set would avoid eliciting a broad though often poorly characterized immunity traditionally associated with a whole virus or virus-derived immunogens rather a focused and precise way to stimulate a specific immune response. The Invention also provides immune-stimulation to post-exposure cases as well. Administration of these epitope-peptides could limit the spread of viral pathogens and facilitate rapid elimination due to a mAb-like neutralizing effect. Thus, in an embodiment, the Invention provides therapeutics against exposure to KFDV.
In an embodiment, the invention provides 15 T-cell (MHC-I and MHC-II) dominant epitopes and 11 linear and seven discontinuous B-cell epitopes as a probable candidate was used for vaccine designing. This will elicit the antibody production against KFDV, which is also conserved for ALKV and TBEV. Thus, a mixture of these peptides would act as vaccine candidates for all three viruses.

In an embodiment, the Invention provides vaccine comprising immunodominant peptides. Peptides are inert and non-infectious substances and could be synthesized without the involvement of animal tissue culture. The entire process could be performed in regular BSL-2 conditions and there is no need to work with any infectious agent for the formulation.

In an embodiment, the Invention identifies immunogenic peptides and synthesizes the peptides in vitro. Alternatively, the peptides can also be procured from companies. The pure peptides are used for animal (mice and rabbit) studies for testing the produced antibodies' efficiency by neutralization assay with KFDV chimeric virus/VLPs. Produced neutralizing antibodies could be used as therapeutics, and responsible peptides can be used as a vaccine candidate.

KFDV T-cell epitope prediction
For the prediction of MHC-I restricted T cell epitopes for KFDV, the analysis was done with NetMHC 4.0 server (http://www.cbs.dtu.dk/services/NetMHC/). Using NetMHC 4.0 server, the Inventors predicted CD8+ T-cell epitopes against 36 HLA-A, 34 HLA-B, 10 HLA-C, and 1 HLA-E (which included 12 HLAs super types A1, A2, A3, A24, A26, B7, B8, B27, B39, B44, B58, and B62) alleles. The peptide length was set to nine amino acids. The potent binding epitopes was selected based on the threshold-strong binding % rank cut-off value as 0.5, and the threshold weak binder % rank cut-off as 2.0. The binding affinity was set at = 50nM.

Similarly, KFDV envelope protein T-cell MHC-II epitopes were predicted using NetMHCIIpan 4.0 Server (http://www.cbs.dtu.dk/services/NetMHCIIpan/). Using NetMHCIIpan 4.0 server, the Invention predicts CD4 T-cell epitopes against 607 DRB1, 32DRB3, 6DRB4, 15 DRB5, 2048 DP, and 2912 DQ alleles. The peptide length was set to 15 amino acids. The potent binding epitopes was selected based on the threshold-strong binding % rank cut-off value as 5, and the threshold weak binder % rank cut-off as 2.0. The binding affinity was set at = 50nM.

The obtained KFDV MHC-I and MHC-II epitopes were clustered using the IEDB tool Epitope cluster analysis server (http://tools.iedb.org/cluster/). Epitopes were clustered using a minimum sequence identity threshold of 70%, and conserved peptides were clustered using cluster-break for a precise representative sequence. The resulted epitopes were further sorted by antigenicity, immunogenicity, allergenicity, toxicity, and conservancy scores.
The antigenicity of clustered epitopes was calculated using VaxiJen v2.0 server (http://www.ddg-pharmfac.net/vaxijen/VaxiJen/VaxiJen.html). The clustered MHC-I CD8 epitopes immunogenicity predicted using IEDB class-I immunogenicity prediction tool (http://tools.iedb.org/immunogenicity/). The Invention follows the recommendations from IEDB for threshold settings (PMID: 28352270). The conservancy of MHC-I, and MHC-II epitopes was calculated in 56 KFDV serotypes using the IEDB epitope conservancy analysis tool (http://tools.iedb.org/conservancy/). The conservancy analysis type was selected as Epitope linear sequence conservancy, and the sequence identity threshold is = 90%.
The toxicity of MHC-I and MHC-II predicted peptides was calculated using ToxinPred (http://www.imtech.res.in/raghava/toxinpred/) webserver. Allergenicity of predicted MHC-I and MHC-II peptides were predicted AllergenFP 1.0 (http://www.ddg-pharmfac.net/AllergenFP/) server. IEDB population coverage tool (http://tools.iedb.org/population/) was used for the analysis of the population coverage of the predicted CD4 and CD8 cell epitopes and their respective MHC HLA-binding alleles.

Table 1: KFDV MHC-I and MHC-II dominant epitopes.
KFDV MHC-Iz
Epitopes HLA alleles Start -End
TRVSLVLEL HLA-B*39:01 19-27
LTAEGKPSV HLA-A*69:01 33-41
KTREYCLHAKL HLA-A*30:01,HLA-B*40:01,HLA-B*40:02 55-65
CPAMGPATL HLA-B*07:02,HLA-B*35:01,HLA-B*07:02 74-82
KKKATGYVY HLA-B*15:03 124-132
HSNRKTASF HLA-B*15:03,HLA-B*15:17,HLA-B*58:01 157-165
LPWRHGGAQ HLA-B*07:02 225-233
QEWNHADRL HLA-B*40:01 233-241
METTGGGFV HLA-B*40:01 364-372
YVGELSHQW HLA-B*58:01 384-392
SVGGMLSSV HLA-A*02:03 425-433
ALHTAFGAAF HLA-A*02:03,HLA-B*15:03 436-445
LVLTMTLGV HLA-A*02:01,HLA-A*02:06 486-494
KFDV (MHC-II)
VRAVAHGEPNVNVAS HLA-DQA1*05:05/DQB1*03:13, HLA-DQA1*05:04/DQB1*03:13, HLA-DQA1*05:03/DQB1*03:13, HLA-DQA1*05:01/DQB1*03:13, HLA-DQA1*02:01/DQB1*03:13, HLA-DQA1*05:06/DQB1*03:13, HLA-DQA1*05:07/DQB1*03:13, HLA-DQA1*05:08/DQB1*03:13, HLA-DQA1*05:09/DQB1*03:13, HLA-DQA1*05:10/DQB1*03:13, HLA-DQA1*05:11/DQB1*03:13 342-356
VGGMLSSVGKALHTA HLA-DRB1*14:74, HLA-DRB1*09:06 426-440

Table 2: Population coverage of epitopes and their respective HLAs.
KFDV
Population Coveragea Averageb pc90c
East Asia 77.34% 1.55 0.44
Japan 74.42% 1.46 0.39
Northeast Asia 76.73% 1.72 0.43
China 75.82% 1.64 0.41
South Asia 76.49% 1.33 0.43
Sri Lanka 18.46% 0.24 0.12
Pakistan 32.18% 0.35 0.15
India 75.39% 1.31 0.41
Southeast Asia 77.04% 1.99 0.44
Singapore 59.24% 1.43 0.25
Southwest Asia 85.69% 1.64 0.7
Saudi Arabia 59.09% 1.07 0.24
Europe 91.86% 2.21 1.07
Italy 95.36% 2.31 1.22
Czech Republic 92.06% 2.19 1.08
Germany 90.3% 2.2 1.01
England 90.17% 2.28 1.01
Russia 87.34% 1.99 0.79
United Kingdom 26.9% 0.27 0.14
East Africa 76.36% 1.66 0.42
Uganda 76.3% 1.72 0.42
West Africa 88.24% 1.97 0.85
Central Africa 67.89% 1.3 0.31
North Africa 88.53% 1.92 0.87
South Africa 42.03% 0.92 0.17
West Indies 82.74% 1.79 0.58
North America 90.78% 2.18 1.03
United States 93.64% 2.33 1.14
Central America 78.52% 0.88 0.47
South America 73.89% 1.23 0.38
Oceania 72.55% 1.38 0.36

a Projected population coverage.
b Average number of epitope hits/HLA combinations recognized by the population.
c Minimum number of epitope hits/HLA combinations recognized by 90% of the population.

KFDV envelope protein structure modeling
Identifying proper B-cell epitopes is central to any vaccine designing as priming of B cells, and subsequent development of protective humoral immune response is centered around this. Among the two forms, the discontinuous B cell epitopes are the majority form, which comprises discrete amino acid residues arranged together as per the protein folding and 3D structure. While dimeric, trimeric, and antibody-bound dimeric crystal and cryo-EM structures are available TBEV envelope protein in Protein Data Bank (PDB), the same is lacking for KFDV. Hence homology modeling-based protein structure prediction was made for KFDV (UniProtKB ID: D7RF80) Envelope protein. The 3D-structure was modeled around the closest (TBEV: 82.4% homology) structure using I-TASSER (https://zhanglab.ccmb.med.umich.edu/cgi-bin/itasser_submit.cgi). Models were generated using the threading approach, and the best model was selected based on C-score (Table-3). The resultant 3D-monomeric structure was refined using ModRefiner (https://zhanglab.ccmb.med.umich.edu/ModRefiner/), and the quality of structure was validated using PROCHECK. The Ramachandran plot parameter were presented in the Table-3. The biological dimer and trimer were generated by symmetry operation in Pymol.

Table 3: The KFDV structure modeling parameters.
KFDV (Dimer) KFDV (Trimer)
C-Score 2 -1.73
TM Score 0.99±0.04 0.50+-0.15
RMSD 3.1±2.2Å 11.4+-4.5
Most favored regions 85.9% 84.3%
Additional allowed region 11% 12.7%
Generously allowed region 1.7% 2.4%
Disallowed 1.4% 0.6%

The modeled KFDV envelope protein dimer structures resembled other flaviviruses envelope protein structures. The envelope protein contains ~500 aa, where the N-terminal 400 residues form the soluble ectodomain while the rest ~100 C-terminal residues formed the flexible stem and transmembrane domain. The soluble ectodomain has three subdomains; briefly, domain-I is a central ß-barrel domain formed by three stretches of amino acids 1-51, 132-195, and 280-295, and is linking between the domain-II and domain-III (Fig-1). Fig. 1 illustrates the upper panel which is a dimeric monomer (left) and dimer (right). The domains are represented in D1 Bricred color, D2-yellow, D3-Cyan, and D4-Deep blue. The lower pane is a trimeric monomer (left) and trimer (right). The trimer subunit-A is Magenta color, Subunit-B is blue, and Subunit-C is golden color. A flexible hinge connects the domain-I and domain-II. The domain-II is an extended dimerization domain formed by two stretches of amino acids 52-131 and 196-279. While domain-III is formed in a single stretch of residues 296-397 folded like standard immunoglobulin domain (Fig-1). The flexible domain I-II linker confer conformational changes associated with virus maturation and fusion. The C-terminal domain or domain-IV is formed by aa 398-492 in four alpha-helices; the first two a-helices are amphipathic and have a flat orientation on the viral envelope. In comparison, the third and fourth a-helices are antiparallel and are inserted into the viral membrane envelope. Two envelope protein subunits form the flavivirus mature/metastable dimers in an antiparallel fashion. Additionally, two M-proteins units bind with the transmembrane domain resulting in the whole complex to a dimer of dimers (two envelope and two M proteins).
KFDV envelope protein B-cell linear and discontinuous epitope prediction
It has been reported that >90% of B-cell epitopes are discontinuous, i.e., the discontinuous epitope consists of segments distantly separated in the protein sequence that is closely represented by the protein folding. The remaining 10%, B-cell present the linear sequences as epitopes. The Inventors predicted B-cell linear epitopes using IEDB tolls BepiPred-2.0 (http://tools.iedb.org/bcell/), and discontinuous epitopes were predicted using IEDB DiscoTope 2.0 servers, which are embedded in the IEDB database. BepiPred-2.0 was developed based on the random forest algorithm trained on epitopes annotated from antigen-antibody protein structures.

Table 4: KFDV B-cell linear epitopes
Epitopes KFDV
Epitope-1 5HLQNR9
Epitope-2 48IHQENPAKT56
Epitope-3 68SKVAARCPAMGPATLPEEHQASTVCRRDQSDRGWGN103
Epitope-4 148GDYLAANESHSNR160
Epitope-5 191GVD193
Epitope-6 204KTAEHLPKA212
Epitope-7 218DWFEDLSLPWRHGGAQEWNHAD239
Epitope-8 311KFAWKRPPTDSG322
Epitope-9 348GEPN351
Epitope-10 391HQWFQKGST398
Epitope-11 420AWDFGSVG427

To compare the dimeric and trimeric epitopes, the Inventors used KFDV (dimer, trimer) for the prediction of conformational epitopes. The Inventors have taken on an average cut-off DiscoTope score is - 4.0. The dimeric and trimeric protein epitope residues within the cut-off range are rendered on 3D-structures (Fig-2). Fig-2 illustrates KFDV envelope protein conformational epitopes. Upper panel is KFDV dimer cartoon structure depicted in purple color monomer (left) dimer (right) epitope amino acid showed as spheres in the slate color. The lower panel is KFDV trimeric cartoon structure depicted in purple color for monomer (left) trimer (right) epitope amino acid spheres in ruby color. 3D-rendering was performed using Pymol.
The close contact residues are grouped as a one discontinuous epitope Table-5. Like that, the Inventors grouped seven epitopes on KFDV dimer and five epitopes on trimer conformation.

Table-5. KFDV, ALKV, and TBEV envelope protein conformational epitopes
Epitopes KFDV (Dimer) KFDV (Trimer)
Epitope-1 100G, 101W, 102G, 108F 100G, 101W, 102G, 108F
Epitope-2 230G, 231G 231G
Epitope-3 51E, 52N, 54A, 277E, 278G, 279S, 280K 51E, 52N, 277E, 278G, 279S, 280K
Epitope-4 154N 15T, 16Q, 148G, 149D, 150Y, 151L, 152A, 153A
Epitope-5 178D, 366T, 367T, 368G -
Epitope-6 302M, 303T, 335S, 336K, 337P -
Epitope-7 347H, 348G, 349E, 350P, 378P, 379G, 380D, 391Q, 393F, 395K, 396G, 397S, 398T, 399I, 400G 348G, 349E, 350P, 391Q
,CLAIMS:We Claim:
1. A Kyasanur Forest Disease Virus envelope protein peptide-based vaccine comprising:
c) 15 T-cell comprising MHC-I and MHC-II, dominant epitopes;
d) 11 linear and seven discontinuous B-cell epitopes

2. The vaccine as claimed in claim 1, is effective against ALKV and TBEV.
3. The vaccine as claimed in claim 1, wherein KFDV MHC-I dominant epitopes comprise:
KFDV MHC-I
Epitopes HLA alleles Start -End
TRVSLVLEL HLA-B*39:01 19-27
LTAEGKPSV HLA-A*69:01 33-41
KTREYCLHAKL HLA-A*30:01,HLA-B*40:01,HLA-B*40:02 55-65
CPAMGPATL HLA-B*07:02,HLA-B*35:01,HLA-B*07:02 74-82
KKKATGYVY HLA-B*15:03 124-132
HSNRKTASF HLA-B*15:03,HLA-B*15:17,HLA-B*58:01 157-165
LPWRHGGAQ HLA-B*07:02 225-233
QEWNHADRL HLA-B*40:01 233-241
METTGGGFV HLA-B*40:01 364-372
YVGELSHQW HLA-B*58:01 384-392
SVGGMLSSV HLA-A*02:03 425-433
ALHTAFGAAF HLA-A*02:03,HLA-B*15:03 436-445
LVLTMTLGV HLA-A*02:01,HLA-A*02:06 486-494

4. The vaccine as claimed in claim 1, wherein KFDV MHC-II dominant epitopes comprise:
KFDV MHC-II
Epitopes HLA alleles Start -End
VRAVAHGEPNVNVAS HLA-DQA1*05:05/DQB1*03:13, HLA-DQA1*05:04/DQB1*03:13, HLA-DQA1*05:03/DQB1*03:13, HLA-DQA1*05:01/DQB1*03:13, HLA-DQA1*02:01/DQB1*03:13, HLA-DQA1*05:06/DQB1*03:13, HLA-DQA1*05:07/DQB1*03:13, HLA-DQA1*05:08/DQB1*03:13, HLA-DQA1*05:09/DQB1*03:13, HLA-DQA1*05:10/DQB1*03:13, HLA-DQA1*05:11/DQB1*03:13 342-356
VGGMLSSVGKALHTA HLA-DRB1*14:74, HLA-DRB1*09:06 426-440

5. The vaccine as claimed in claim 1, wherein the KFDV B-cell linear epitopes comprises:
Epitopes KFDV
Epitope-1 5HLQNR9
Epitope-2 48IHQENPAKT56
Epitope-3 68SKVAARCPAMGPATLPEEHQASTVCRRDQSDRGWGN103
Epitope-4 148GDYLAANESHSNR160
Epitope-5 191GVD193
Epitope-6 204KTAEHLPKA212
Epitope-7 218DWFEDLSLPWRHGGAQEWNHAD239
Epitope-8 311KFAWKRPPTDSG322
Epitope-9 348GEPN351
Epitope-10 391HQWFQKGST398
Epitope-11 420AWDFGSVG427