Molecular modeling andin-silicoengineering ofCardamom mosaic viruscoat protein for the presentation of immunogenic epitopes ofLeptospiraLipL32

Abstract

Expression of Cardamom mosaic virus (CdMV) coat protein (CP) in E. coli forms virus-like particles. In this study, the structure of CdMV CP was predicted and used as a platform to display epitopes of the most abundant surface-associated protein, LipL32 of Leptospira at C, N, and both the termini of CdMV CP. In silico, we have mapped sequential and conformational B-cell epitopes from the crystal structure of LipL32 of Leptospira interrogans serovar Copenhageni str. Fiocruz L1-130 using IEDB Elipro, ABCpred, BCPRED, and VaxiJen servers. Our results show that the epitopes displayed at the N-terminus of CdMV CP are promising vaccine candidates as compared to those displayed at the C-terminus or at both the termini. LipL32 epitopes, EP2, EP3, EP4, and EP6 are found to be promising B-cell epitopes for vaccine development. Based on the type of amino acids, length, surface accessibility, and docking energy with CdMV CP model, the order of antigenicity of the LipL32 epitopes was found to be EP4 > EP3 > EP2 > EP6.

Analytical evaluation of an immunomagnetic separation PCR assay to detect pathogenic Leptospira in cattle urine samples obtained under field conditions

“…In order to produce polyclonal antibodies against LipL32 (an outer membrane lipoprotein 7 ), 3 peptide chains (EP3, EP4, and EP6), 9 each with a size of ~20 kDa, were chemically synthesized (this mixture hereafter referred to as P2). Sequences of these LipL32 epitopes have been described in silico as the most immunogenic and are found outside the leptospiral bacterial cell wall.…”

“…Sequences of these LipL32 epitopes have been described in silico as the most immunogenic and are found outside the leptospiral bacterial cell wall. 9 A glutaraldehyde cross-linking protocol with hemocyanin carrier protein (Imject Blue carrier protein; Thermo Fisher Scientific) was performed on the peptides, as described previously, 11 with some modifications. Briefly, 10 mg of hemocyanin was incubated with 1 mg of each peptide in the presence of 0.15% glutaraldehyde (MilliporeSigma) at room temperature for 2 h. The reaction was terminated by the addition of 1/10 (v/v) of 1 M glycine, and the sample was dialyzed against 0.1 M borate buffer (18.55 g of boric acid; 2,850 mL of H 2 O; pH 8.5) for 24 h at 4°C.…”

Molecular Modelling and Insilico Engineering of PapMV-CP Towards Display and Development of Capripox Viral Like Particles Based on Immunogenic P32 Envelop Protein is the Homologous of the Vaccinia-Viral H3L Gene: An Insilico Approach

“…HADDOCK web server clustered the structures and the top ten were analyzed as they have the most reliable structural interaction based on the score obtained by each cluster. The epitopes when individually docked at N and C-terminal region of the PMVCP (Kumar et al 2015). The docked Fig.…”

Future of Cancer Immunotherapy using Plant virus-based Nanoparticles

“…[47] used a mathematical model to explore the diffusion and cellular uptake of TMV in the form of a spheroid system rather than its native rod shape, in the presence and absence of modified surface chemistry. A combination of in silico and wet lab experiments can assist in the rational design of newly engineered VLPs that carry immunogenic peptides specific for tumors [48]. Alternatively, computational modeling can help design plant virus nanoparticles to be carriers for anticancer drugs [33,49].…”