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MALDI-TOF MS of preparations of peptide Biotinyl-SGSGNTKLMGGT-(KP) (mol. mass 1542.3) obtained with the help of modifications (a), (b), (c), and (d) of the standard schedule of parallel peptide synthesis on pins (see Section 2 for details).
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Unified schedule for multiple parallel solid-phase synthesis of so-called "difficult" peptides on polypropylene pins was developed. Increase in the efficiency of 9-fluorenyl(methoxycarbonyl) N-terminal amino-protecting group removal was shown to have a greater influence on the accuracy of the "difficult" peptide synthesis than the use of more effic...
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... For that reason, a multi-epitope-based peptide vaccine can be designed with immunogenic proteins of a pathogen. In the present scenario, a large number of peptide vaccines are in progress, majority of which are for human infectious diseases and tumor (55)(56)(57)(58). Very few studies are reported in the field of in silico vaccine for poultry and animals. ...
Avian mycoplasma is a bacterial disease causing chronic respiratory disease (CRD) in poultry industries with high economic losses. The eradication of this disease still remains as a challenge. A multi-epitope prophylactic vaccine aiming the antigenic proteins of Mycoplasma gallisepticum can be a capable candidate to eradicate this infection. The present study is focused to design a multi-epitope vaccine candidate consisting of cytotoxic T-cell (CTL), helper T-cell (HTL), and B-cell epitopes of antigenic proteins, using immunoinformatics strategies. The multi-epitopic vaccine was designed, and its tertiary model was predcited, which was further refined and validated by computational tools. After initial validation, molecular docking was performed between multi-epitope vaccine construct and chicken TLR-2 and 5 receptors, which predicted effective binding. The in silico results specify the structural stability, precise specificity, and immunogenic response of the designed multi-epitope vaccine, and it could be an appropriate vaccine candidate for the M. gallisepticum infection.
... Similar to succinylation and malonylation, the glutarylation of protein lysine residues not only increases the length was glutarylated on the matrix with glutaric anhydride in DMF in the presence of Nethyl-N,N-di-iso-propylamine. Fmoc-deprotection was performed with the solution of 2% 4-methylpiperidine and 2% 1,8-diazabicyclo [4,5,0] undec-7-ene in DMF [23]. Peptides were cleaved from the matrix with a TFA-water (19:1, v/v) mixture and precipitated with ether. ...
The glutarylation of lysine residues in proteins attracts attention as a possible mechanism of metabolic regulation, perturbed in pathologies. The visualization of protein glutarylation by antibodies specific to ε-glutaryl-lysine residues may be particularly useful to reveal pathogenic mutations in the relevant enzymes. We purified such antibodies from the rabbit antiserum, obtained after sequential immunization with two artificially glutarylated proteins, using affinity chromatography on ε-glutaryl-lysine-containing sorbents. Employing these anti(ε-glutaryl-lysine)-antibodies for the immunoblotting analysis of rat tissues and mitochondria has demonstrated the sample-specific patterns of protein glutarylation. The study of the protein glutarylation in rat tissue homogenates revealed a time-dependent fragmentation of glutarylated proteins in these preparations. The process may complicate the investigation of potential changes in the acylation level of specific protein bands when studying time-dependent effects of the acylation regulators. In the rat brain, the protein glutarylation, succinylation and acetylation patterns obtained upon the immunoblotting of the same sample with the corresponding antibodies are shown to differ. Specific combinations of molecular masses of major protein bands in the different acylation patterns confirm the selectivity of the anti(ε-glutaryl-lysine)-antibodies obtained in this work. Hence, our affinity-purified anti(ε-glutaryllysine)-antibodies provide an effective tool to characterize protein glutarylation, revealing its specific pattern, compared to acetylation and succinylation, in complex protein mixtures.
... The presence of lysine allows this peptide to be acylated at its less hydrophobic C-terminus with a lipophilic adjuvant through the ε-amino group of lysine, provided it is blocked by a selectively removable protective group. The peptide was obtained by solid-phase synthesis on Rink amide resin according to the Fmoc protocol by the method of in situ activated esters [28,29]. After synthesis, the peptide was purified by reversed phase high-performance liquid chromatography (RP-HPLC) using UV and mass-spectrometric detection to identify the target peptide and impurities. ...
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The work is aimed at the synthesis and analysis from NS4A of hepatitis C virus (HCV) antigen peptide fragment that contains a conserved B-cell and T-helper epitopes. The 24-mer peptide VIVGRIILSGRPAVIPDREVLYRK-NH2, which contains the main immunogenic site 24–46 of HCV NS4A antigen (corresponding to the 1681–1703 amino acid residues of the HCV polypeptide), subtype 1b, has been prepared via solid-phase synthesis according to the Fmoc-protocol. Particles with diameters of 73 ± 10 nm (30%) and 236 ± 5 nm (70%) have been detected in the water solution of the highly purified peptide (0.5 mg/mL) by dynamic light scattering. The polydispersity index of 0.377 ± 0.012 implies the existence of heterogeneity because of the aggregation of the peptide molecules. The ζ-potential of the peptide aggregates has been determined as 7.0 ± 0.5 mV by means of electrophoretic light scattering. These data confirm the possibility for the development of a nanoscale liposome form of the peptide preparation. Immunoreactivity of the synthesized highly purified peptide has been studied with the use of blood sera of patients with chronic hepatitis C. Antipeptide immunoglobulins G have been detected in 41.7% of serum samples. Thus, this peptide has been shown to reproduce at least one B-epitope, to which antibodies are raised during natural HCV infection. The synthesized 24-mer peptide is a promising candidate for further research and for use as a potential immunogen for the design of a nanoscale therapeutic immunogenic liposomal peptide composition with synthetic lipids as an adjuvant.
... For development of B-Cell responses, Virus like Particles (VLP) or nanoparticles are linked to the selected peptide so that the presentation of ordered, multivalent epitopes can efficiently cross-link B cell receptors (BCRs), required for improved antibody response [34]. Antibodies can target the specific epitopes in conformation dependent or independent manners (Linear epitopes) [35]. Protein or peptide engineering is used to mimic the "structural epitope" or conformation dependent larger epitopes [36,37]. ...
RNA viruses have high rate of replication and mutation that help them adapt and change according to their environmental conditions. Many viral mutants are the cause of various severe and lethal diseases. Vaccines, on the other hand have the capacity to protect us from infectious diseases by eliciting antibody or cell-mediated immune responses that are pathogen-specific. While there are a few reviews pertaining to the use of artificial intelligence (AI) for SARS-COV-2 vaccine development, none focus on peptide vaccination for RNA viruses and the important role played by AI in it. Peptide vaccine which is slowly coming to be recognized as a safe and effective vaccination strategy has the capacity to overcome the mutant escape problem which is also being currently faced by SARS-COV-2 vaccines in circulation.Here we review the present scenario of peptide vaccines which are developed using mathematical and computational statistics methods to prevent the spread of disease caused by RNA viruses. We also focus on the importance and current stage of AI and mathematical evolutionary modeling using machine learning tools in the establishment of these new peptide vaccines for the control of viral disease.
... On the contrary, because of the small size peptide vaccine is weakly immunogenic and require a carrier molecule for adjuvanting and chemical stability [19]. Several peptide vaccines are under process of development for different viral diseases, such as hepatitis C virus (HCV) [23], human papilloma virus (HPV) [24], human immunodeficiency virus (HIV) [25], and influenza [26] etc. A multi-epitope vaccine contains a series of overlapping epitopes that can elicit effective cytotoxic T cells, T helper cells and B cells responses against viral pathogen [27]. ...
Middle East respiratory syndrome coronavirus (MERS-CoV) is an emerging health concern due to its high mortality rate of 35%. At present, no vaccine is available to protect against MERS-CoV infections. Therefore, an in silico search for potential antigenic epitopes in the non-redundant proteome of MERS-CoV was performed herein. First, a subtractive proteome-based approach was employed to look for the surface exposed and host non-homologous proteins. Following, immunoinformatics analysis was performed to predict antigenic B and T cell epitopes that were used in the design of a multi-epitopes peptide. Molecular docking study was carried out to predict vaccine construct affinity of binding to Toll-like receptor 3 (TLR3) and understand its binding conformation to extract ideas about its processing by the host immune system. We identified membrane protein, envelope small membrane protein, non-structural protein ORF3, non-structural protein ORF5, and spike glycoprotein as potential candidates for subunit vaccine designing. The designed multi-epitope peptide then linked to β-defensin adjuvant is showing high antigenicity. Further, the sequence of the designed vaccine construct is optimized for maximum expression in the Escherichia coli expression system. A rich pattern of hydrogen and hydrophobic interactions of the construct was observed with the TLR3 allowing stable binding of the construct at the docked site as predicted by the molecular dynamics simulation and MM-PBSA binding energies. We expect that the panel of subunit vaccine candidates and the designed vaccine construct could be highly effective in immunizing populations from infections caused by MERS-CoV and could possible applied on the current pandemic COVID-19.
... Therefore epitopes/peptide based vaccines can be an alternative strategy for an efficient vaccine production, to induce positive humoral and cell mediated immune responses (Sesardic, 1993;Li et al., 2014). There are many peptide based vaccines which are under development to combat several disease, such for anthrax (Oscherwitz et al., 2010), hepatitis C virus (HCV) (Kolesanova et al., 2013), malaria (Epstein et al., 2007)], foot and mouth disease (FMD) (Volpina et al., 1993), swine fever (Tarradas et al., 2011), influenza (Stanekova et al., 2011), human immunodeficiency virus (HIV) (Liu et al., 2007), human papilloma virus (HPV) (Solares et al., 2011). ...
... A large amount of research is being focused toward developing peptide vaccines against different disease. To-date, a large amount of peptide vaccines has either been developed or are under development against influenza [70], various types of cancer [71][72][73], hepatitis C virus (HCV) [74], human papilloma virus (HPV) [75], malaria [76], swine fever [77], human immunodeficiency virus (HIV) [78], foot and mouth disease [79], anthrax [80] etc. Several peptide vaccines are available to treat neurodegenerative disorders as well. ...
Bioactive peptides are actively involved in different biological functions and importantly contribute to human health, and the use of peptides as therapeutics has a long successful history in disease management. A number of peptides have wide-ranging therapeutic effects, such as antioxidant, antimicrobial, and antithrombotic effects. Neurodegenerative diseases are typically caused by abnormal aggregations of proteins or peptides, and the depositions of these aggregates in or on neurons, disrupt signaling and eventually kill neurons. During recent years, research on short peptides has advanced tremendously. This review offers a brief introduction to peptide based therapeutics and their application in disease management and provides an overview of peptide vaccines, and toxicity related issues. In addition, the importance of peptides in the management of different neurodegenerative diseases and their therapeutic applications is discussed. The present review provides an understanding of peptides and their applications for the management of different diseases, but with focus on neurodegenerative diseases. The role of peptides as anti-cancer, antimicrobial and antidiabetic agents has also been discussed.
... Peptide vaccines under various phases of trial and development, the vast majority of them related to cancers. [52][53][54][55][56][57][58][59][60][61][62][63][64]. ...
Merkel cell Polyomavirus is non-enveloped, dsDNA virus belonging to Polyomaviridae family linked to an
uncommon aggressive skin malignancy. The poor prognosis and limited understanding of disease pathogenesiswarrants innovative treatment. In this current study we aim to predict TB cell immunogenic epitopes from theVP1 protein of all merkel cell polyomavirus strain which will aid in effective epitope based vaccine design usingimmuoinformatics approaches. We retrieved 423 full-length VP1 protein sequences of merkel cell polyomavirusevirus species from the NCBI database. These sequences were analyzed to determine the conserved region and were used to predict the epitopes using the IEDB immunoinformatics algorithms. For B cell three epitope were predictedas peptide vaccine (QEKTVY, KTVYPK, and QEKTVYP). For T cell the predicted Class-I peptides (SLFSNLMPK, LQMWEAISV and LLVKGGVEV) were found to cover the maximum number of MHC I alleles. The highest scoringClass II MHC binding peptides were (IELYLNPRM, ISSLINVHY and INSLFSNLM). Further experiments will needto be undertaken to confirm the potential of these predicted epitopes in a future efficacious vaccine development.
... Peptide vaccination is a key role of combining a good desirable immune response and a minimal immunological side effect. There are many peptide vaccines under development, such as vaccine for human immunodeficiency virus (HIV), hepatitis C virus (HCV), malaria, foot and mouth disease, swine fever, influenza, anthrax, human papilloma virus (HPV), therapeutic anti-cancer vaccines, pancreatic cancer, melanoma, non-small cell lung cancer, advanced hepatocellular carcinoma, cutaneous T-cell lymphoma and B-Cell chronic lymphocytic leukaemia [54][55][56][57][58][59][60][61][62][63][64][65][66][67]. ...
Sudan Ebola virus is single stranded negative sense RNA genome belonging to Filovirus Filoviridae family that causes hemorrhagic fever. There is no treatment or vaccine for it, thus the aim of this study is to design a peptide vaccine using immuoinformatics approaches to analyse the glycoprotein of the all strain of SUDV, to determine the conserved region which is further studied to predict all possible epitopes that can be used as a peptide vaccine. A total of 21 Sudan Ebola virus glycoprotein retrieved from NCBI database were aligned to determine the conservancy and to predict the epitopes using IEDB analysis resource. Three epitopes predicted as a peptide vaccine for B cell (PPPPDGVR, ETFLQSPP, LQSPPIRE). For T cell four epitopes showed high affinity to MHC class I (FLYDRLAST, IIIAIIALL, MHNQNALVC and RTYTILNRK) and high coverage against Sudan and the whole world population. Also in MHC class II, Four epitopes that interact with most frequent MHC class II alleles (FAEGVIAFL, FLRATTELR, FLYDRLAST and FVWVIILFQ) with high coverage against Sudan and the whole world population. We recommend in vivo and in vitro study to prove the effectiveness of these predicted epitopes as a peptide vaccine.
... From crystallographic studies, it was found that the HLA-DM crystal is topologically similar to HLA-DR molecules, but lacks a peptide-binding cleft (84,140). Recently, it was found that HLA-DM catalyzes MHC class II peptide loading by targeting the P1-pocket (141)(142)(143). The aW43 amino acid of HLA-DR is rotated outside from the cleft and makes H-bonds with the aN125 amino acid of HLA-DM and forms the HLA-DM-DR complex. ...
Major histocompatibility complex (MHC) class II molecules are expressed by antigen-presenting cells and stimulate CD4(+) T cells, which initiate humoral immune responses. Over the past decade, interest has developed to therapeutically impact the peptides to be exposed to CD4(+) T cells. Structurally diverse small molecules have been discovered that act on the endogenous peptide exchanger HLA-DM by different mechanisms. Exogenously delivered peptides are highly susceptible to proteolytic cleavage in vivo; however, it is only when successfully incorporated into stable MHC II-peptide complexes that these peptides can induce an immune response. Many of the small molecules so far discovered have highlighted the molecular interactions mediating the formation of MHC II-peptide complexes. As potential drugs, these small molecules open new therapeutic approaches to modulate MHC II antigen presentation pathways and influence the quality and specificity of immune responses. This review briefly introduces how CD4(+) T cells recognize antigen when displayed by MHC class II molecules, as well as MHC class II-peptide-loading pathways, structural basis of peptide binding and stabilization of the peptide-MHC complexes. We discuss the concept of MHC-loading enhancers, how they could modulate immune responses and how these molecules have been identified. Finally, we suggest mechanisms whereby MHC-loading enhancers could act upon MHC class II molecules.
