Novel Multi-Epitope Vaccine candidate derived from critically important proteins of Monkeypox isolates

Abstract

Background: On May 7, 2022, a case of monkeypox virus (MPXV) has been reported to the WHO. It causes a viral zoonotic disease with characteristics comparable to that of smallpox cases. Monkeypox could be a serious health concern as it is already spreading to multiple countries worldwide. To prevent the monkeypox infection and for its early detection, a potential vaccine and diagnostic candidate is urgently required. Method: In present study we have used different in silico methods to screen the entire genome with its approximately 191 genes for potential epitopes present in proliferation and virulent proteins of monkeypox virus. All protein sequences were retrieved from different genomic and proteome databases listed in Uniprot or NCBI. Results: In the present study we have screened potential epitopes from 11 different proteins of Monkeypox. All the included protein play an important role in pathogenesis and/or proliferation of Monkeypox virus. We have identified in total 984 CTL and 168 HTL epitopes with highest score in our epitope screening. The reported epitopes could be potential candidates for the design of an early detection diagnostic kit specific for the monkeypox virus. Out of these target peptides we have included a total of 39 CTL epitopes and 39 HTL epitopes in design of multi-epitope vaccine candidates. These shortlisted epitopes are highly conserved amongst different strains and origin of monkeypox viruses. The population coverage by joint administration of CTL and HTL MEVs is predicted to be high with the epitopes showing potential to bind upto 24 different CTL and HTL HLA allele molecules. The epitopes used in MEVs are examined to be highly immunogenic, non allergic but antigenic, and non toxic. All the CTL and HTL MEVs designed utilizing the epitopes have physiochemical properties favor its over expression in human cells. The optimized cDNA constructs of CTL and HTL MEVs also favor over expression of MEVs in human cells. Overall, the MEV construct proposed by us are fissile for expression in the lab and for further in vivo studies. Conclusion: Control and fight against emerging diseases such as MPXV requires pathogen diagnostic and novel vaccine approaches. We screened for several epitopes and designed a MEV providing a potential solution for both purposes. Our method allows rapid screening and provides a rational strategy for the development of vaccine candidate effective in fighting MPXV and other unexpected upcoming diseases.

Novel “GaEl Antigenic Patches” Identified by a “Reverse Epitomics” Approach to Design Multipatch Vaccines against NIPAH Infection, a Silent Threat to Global Human Health

“…A total of 30 GaEl Ag-Patches from 362 CTL and 27 GaEl Ag-Patches from 414 HTL overlapping epitopes were identified. To identify the GaEl Ag-Patches, a novel “reverse epitomics” approach involving the “overlapping-epitope-clusters-to-patches” method was utilized − (Tables and ). The herewith suggested GaEl Ag-Patches are expected to result in up to 776 overlapping epitopes upon intercellular proteolytic chop down by antigen presenting cells (APCs).…”

“…The constructs include hBD-2 with sequence: GIGDPVTCLKSGAICHPVFCPRRYKQIGTCGLPGTKCCKKP and 3D structure deposited in the Protein Data Bank (PDB code 1FD3) and hBD-3 with sequence: GIINTLQKYYCRVRGGRCAVLSCLPKEEQIGKCSTRGRKCCRRKK and 3D structure (PDB code 1KJ6). hBD 2 and 3 are utilized here as adjuvants to enhance the immunogenic response. − ,,− The multipatch vaccine design and the method thereof are included in the field patents 202011037939, 202011037939, PCT/IN2021/050841, and previous publications. − …”

“…These vaccines are composed of “GaEL antigenic patches” (GaEl Ag-Patches) identified from the proteome of NiV. The “GaEl Ag-Patches” were identified by the reverse epitomics approach as introduced in our earlier studies. − All nine proteins of the NiV proteome () were used in this study, viz., C protein (gi-1859635642), glycoprotein (gi-253559848), fusion protein (gi-13559813), nucleoprotein (gi-1679387250), matrix protein (gi-13559811), phosphoprotein (gi-1802790259), W protein (gi-374256971), V protein (gi-1802790260), and RNA polymerase (gi-15487370). The full-length amino acid sequences of NiV proteins were retrieved from the National Center for Biotechnology Information (NCBI).…”

“…The consensus amino acid sequence of overlapping epitopes was identified as “GaEl Ag-Patches”. This novel approach of search and identification of “GaEl antigenic patches” from a source protein is named as “reverse epitomics” as introduced and explained in our earlier studies. − This approach is applicable to proteins/antigens of any pathogen and not only to SARS-CoV-2 or NiV.…”

“…This is to acknowledge the inventors of “reverse epitomics”, and the prefix GaEl is derived from river Ganga and Elna of the inventors’ home countries. The novel “reverse epitomics” approach to identify “GaEl Ag-Patch” has been introduced in our earlier study on SARS-CoV-2. − In the present study, we have utilized the “reverse epitomics” approach to identify “GaEl Ag-Patch” from NiV proteins. The reverse epitomics approach applies the novel “overlapping-epitope-clusters-to-patches” method to identify the “GaEl Ag-Patches”.…”