A flexible peptide linker enhances the immunoreactivity of two copies HBsAg preS1 (21–47) fusion protein

Hu, Weiguo; Li, Feng; Yang, Xingxiu; Li, Zhen; Xia, Hengchuan; Li, Guangdi; Wang, Yuan; Zhang, Zuchuan

doi:10.1016/j.jbiotec.2003.09.009

Cited by 37 publications

(32 citation statements)

References 19 publications

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“…The peptide linker involving EAAAK has very favourable property to effectively separate bi-functional domains. The peptide linker involving GGGGS allows interaction between domains, hence improving and restoring folding (Arai, Ueda, Kitayama, Kamiya, & Nagamune, 2001;Bai & Shen, 2006;Chen, Zaro, & Shen, 2013;Hajighahramani et al, 2017;Hu et al, 2004;Huston et al, 1988). Truncated Ov-ASP-1 protein (residues 10-153) was used as an adjuvant at N-terminal of MEVs, to enhance the immune response.…”

Section: Selection Of Epitope For Molecular Interaction Study With Hlmentioning

confidence: 99%

Design of novel multi-epitope vaccines against severe acute respiratory syndrome validated through multistage molecular interaction and dynamics

Srivastava

Kamthania

Pandey

et al. 2019

Journal of Biomolecular Structure and Dynamics

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2019)Design of novel multi-epitope vaccines against severe acute respiratory syndrome validated through multistage molecular interaction and dynamics, Journal of Biomolecular Structure and Dynamics, 37:16, 4345-4360, ABSTRACT Severe acute respiratory syndrome (SARS) is endemic in South China and is continuing to spread worldwide since the 2003 outbreak, affecting human population of 37 countries till present. SARS is caused by the severe acute respiratory syndrome Coronavirus (SARS-CoV). In the present study, we have designed two multi-epitope vaccines (MEVs) composed of cytotoxic T lymphocyte (CTL), helper T lymphocyte (HTL) and B cell epitopes overlap, bearing the potential to elicit cellular as well as humoral immune response. We have used truncated (residues 10-153) Onchocerca volvulus activation-associated secreted protein-1 as molecular adjuvants at N-terminal of both the MEVs. Selected overlapping epitopes of both the MEVs were further validated for stable molecular interactions with their respective human leukocyte antigen class I and II allele binders. Moreover, CTL epitopes were further studied for their molecular interaction with transporter associated with antigen processing. Furthermore, after tertiary structure modelling, both the MEVs were validated for their stable molecular interaction with Toll-like receptors 2 and 4. Codon-optimized cDNA of both the MEVs was analysed for their potential high level of expression in the mammalian cell line (Human) needed for their further in vivo testing. Overall, the present study proposes in silico validated design of two MEVs against SARS composed of specific epitopes with the potential to cause a high level of SARS-CoV specific cellular as well as humoral immune response.

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Section: Selection Of Epitope For Molecular Interaction Study With Hlmentioning

confidence: 99%

Design of novel multi-epitope vaccines against severe acute respiratory syndrome validated through multistage molecular interaction and dynamics

Srivastava

Kamthania

Pandey

et al. 2019

Journal of Biomolecular Structure and Dynamics

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“…The α, β, and γ subunits were fused together by linker-peptide to form one protein molecule. The flexible linker peptide was used to link the variable regions of the heavy and the light chain of gene, engineering a recom-binant antibody single-chain Fv [19], and subsequently was widely used in the expression of various recombinant antibodies in immunological researches [20,21]. The designed linker peptide (Gly-Gly-Gly-Gly-Ser-Gly-Gly-Gly-Gly-Ser) had weak interactions with other amino acids within the peptide chain, and thus was considered to aid in the proper folding of the large protein molecule.…”

Section: Discussionmentioning

confidence: 99%

Heterologous expression and characterization of recombinant glycerol dehydratase from Klebsiella pneumoniae in Escherichia coli

Wang

Tian

et al. 2007

Biotechnology Journal

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Glycerol dehydratase (EC 4.2.1.30), as one of the key enzymes in converting glycerol to the valuable intermediate 1,3-propanediol, is important for biochemical industry. The dhaB genes encoding coenzyme B(12)-dependent glycerol dehydratase in Klebsiella pneumoniae were cloned and expressed in Escherichia coli. An effective co-expression system of multiple subunits protein was constructed. Heterologous expression vectors were constructed using the splicing by overlap extension-PCR technique to co-express the three subunits of the glycerol dehydratase. After induction by isopropyl-beta-D-thiogalactopyranoside, SDS-PAGE analysis revealed that: (i) only the alpha subunit of glycerol dehydratase was expressed in direct expression system, (ii) the three subunits of glycerol dehydratase with predicted molecular massess of 64 (agr;), 22 (beta), and 16 kDa (gamma) were expressed simultaneously in co-expression system, and (iii) the fusion expression system expressed the fusion protein of 99 kDa. Enzyme assay showed that the activities of three heterologous expression products were 27.4, 2.3, and 0.2 U/mg. The highest enzyme activity was almost 17 times of that in K. pneumoniae. The recombinant enzyme was purified and biochemically characterized. The apparent Km values of the enzyme for coenzyme B(12) and 1, 2-propanediol were 8.5 nM and 1.2 mM, respectively. The enzyme showed maximum activity at pH 8.5 and 37 degrees C.

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“…Vaccines candidates respectively as adjuvants, by the rigid linker EAAAK favoring domain formation (Hu et al, 2004;Chen et al, 2013;Hajighahramani et al, 2017;Srivastava et al, 2018;Srivastava et al, 2019). COVID-19 patients usually have lung infection and lung infection is usually coupled with increased expression of hBD-2 and hBD-3.…”

Section: Methodsmentioning

confidence: 99%

“…The identified Ag-Patches were highly conserved in nature and were identified on the basis of significant number of overlapping epitope forming clusters (Figure 2, 3 & 4). Ag-Patches together, and to facilitate folding of the protein into its tertiary conformation (Hu et al, 2004;Wilson et al, 2013;Duits et al, 2003;Chen et al, 2013;Hajighahramani et al, 2017;Srivastava et al, 2019;Srivastava et al, 2018;Srivastava et al, 2020a;Srivastava et al, 2020b) ( Figure 1A, 1B, 2 (Table 3). from CTL-MPV-3); likewise, a total of IFN-γ inducing epitopes were screened from two HTL MPVs (83 from HTL-MPV-1 and 98 from HTL-MPV-2).…”

Section: Screening Of Potential Epitopes From Sars-cov-2 Proteomementioning

confidence: 99%

Computationally validated SARS-CoV-2 CTL and HTL Multi-Patch Vaccines designed by reverse epitomics approach, shows potential to cover large ethnically distributed human population worldwide

Srivastava

Verma

Kamthania³

et al. 2020

Preprint

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Background: The SARS-CoV-2 (Severe Acute Respiratory Syndrome Coronavirus 2) is a positive-sense single-stranded RNA coronavirus responsible for the ongoing 2019-2020 COVID-19 outbreak. The highly contagious COVID-19 disease has spread to 216 countries in less than six months. Though several vaccine candidates are being claimed, an effective vaccine is yet to come. In present study we have designed and theoretically validated novel Multi-Patch Vaccines against SARS-CoV-2. Methodology: A novel reverse epitomics approach, overlapping-epitope-clusters-to-patches method is utilized to identify multiple antigenic regions from the SARS-CoV-2 proteome. These antigenic regions are here termed as Ag-Patch or Ag-Patches, for Antigenic Patch or Patches. The identification of Ag-Patches is based on clusters of overlapping epitopes rising from a particular region of SARS-CoV-2 protein. Further, we have utilized the identified Ag-Patches to design Multi-Patch Vaccines (MPVs), proposing a novel methodology for vaccine design and development. The designed MPVs were analyzed for immunologically crucial parameters, physiochemical properties and cDNA constructs. Results: We identified 73 CTL (Cytotoxic T-Lymphocyte), 49 HTL (Helper T-Lymphocyte) novel Ag-Patches from the proteome of SARS-CoV-2. The identified Ag-Patches utilized to design MPVs cover 768 (518 CTL and 250 HTL) overlapping epitopes targeting different HLA alleles. Such large number of epitope coverage is not possible for multi-epitope vaccines. The large number of epitopes covered implies large number of HLA alleles targeted, and hence large ethnically distributed human population coverage. The MPVs:Toll-Like Receptor ectodomain complex shows stable nature with numerous hydrogen bond formation and acceptable root mean square deviation and fluctuation. Further, the cDNA analysis favors high expression of the MPVs constructs in human cell line. Conclusion: Highly immunogenic novel Ag-Patches are identified from the entire proteome of SARS CoV-2 by a novel reverse epitomics approach. We conclude that the novel Multi-Patch Vaccines could be a highly potential novel approach to combat SARS-CoV-2, with greater effectiveness, high specificity and large human population coverage worldwide.

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A flexible peptide linker enhances the immunoreactivity of two copies HBsAg preS1 (21–47) fusion protein

Cited by 37 publications

References 19 publications

Design of novel multi-epitope vaccines against severe acute respiratory syndrome validated through multistage molecular interaction and dynamics

Design of novel multi-epitope vaccines against severe acute respiratory syndrome validated through multistage molecular interaction and dynamics

Heterologous expression and characterization of recombinant glycerol dehydratase from Klebsiella pneumoniae in Escherichia coli

Computationally validated SARS-CoV-2 CTL and HTL Multi-Patch Vaccines designed by reverse epitomics approach, shows potential to cover large ethnically distributed human population worldwide

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