Design, construction and evaluation of multi-epitope antigens for diagnosis of Lyme disease

Abstract

Introduction and objective. Lyme disease (LD) is the most common vector-borne disease in the temperate zone of the Northern Hemisphere. Diagnosis of LD is mainly based on clinical symptoms supported with serology (detection of antiBorrelia antibodies) and is often misdiagnosed in areas of endemicity. Materials and method. In this study, the chimeric proteins (A/C-2, A/C-4 and A/C-7.1) consisting of B-cell epitopes of outer surface proteins OspA and OspC from Borrelia genospecies prevalent in Eastern Slovakia, were designed, over-expressed in E. coli, and used to detect specific anti-Borrelia antibodies in serologically characterized sera from patients with Lyme-like symptoms to evaluate their diagnostic potential. Results. Results showed that chimeras vary in their immuno-reactivity when tested with human sera. Compared with the results obtained from a two-tier test, the application of recombinant multi-epitope chimeric proteins as diagnosis antigens, produced fair agreement in the case of A/C-2 (0.20<κ<0.40) and good agreement (0.60<κ<0.80) when A/C-7.1 was used as capture antigen. Chimera A/C-4 were excluded from further study due to loss of reactivity with OspA-specific antibodies. Conclusions. The combination of specific B-cell epitopes from OspA and OspC proteins may improve the diagnostic accuracy of serologic assays, but further studies are required to address this hypothesis.

Identification of evolutionary conserved DNA sequence and corresponding S21 ribosomal protein region for diagnostic purposes of all Borrelia spirochetes

“…Successful application of chimeric proteins, assembled using molecular biology techniques, in immunological assays for the diagnosis of viral, bacterial and parasitic diseases has already been demonstrated (Alcaro et al, 2003;Holec-Gąsior et al, 2012a, Holec-Gąsior et al, 2012bDrapała et al, 2014;Ferra et al, 2015). To date, only a few studies have shown the reactivity of this kind of proteins with specific IgG antibodies from human sera of individuals with Lyme borreliosis (Gomes-Solecki et al, 2000;Schreterova et al, 2017). However, chimeric proteins are a new generation of recombinant products which have the potential to replace the native antigen (e.i.…”

Recombinant multiepitope proteins expressed in Escherichia coli cells and their potential for immunodiagnosis

“… Recombinant protein E. coli BL21(DE3) cells Indirect ELISA 146 brucellosis-positive serum samples/20 serum samples from healthy individuals/82 serum samples from non-brucellosis diseases Sensitivity: 88.89% Specificity: 85.54% [ 44 ]/China Human Lyme disease/ Borrelia burgdorferi A/C-2, A/C-4, and A/C-7.1 E . coli (XL1-Blue) cells ELISA 169 Lyme-positive serum samples/5 serum samples from healthy individuals A/C-2 Sensitivity: 80.17% Specificity: 52.83% A/C-4 results not shown A/C-7.1 Sensitivity: 91.37% Specificity: 73.58% [ 45 ]/Slovakia Human and goat brucellosis/ Brucella spp. rOmp E. coli BL21(DE3) cells Indirect ELISA 40 human and 31 goat brucellosis-positive serum samples/20 healthy human and 20 healthy goat serum samples rOmp was recognized by human and goat positive samples [ 46 ]/China Goat brucellosis/ Brucella spp.…”

“…Schreterova et al (2017) [ 45 ] conducted a study aimed at developing new RMPs for diagnosing Lyme disease, caused by Borrelia burgdorferi . After using phage display and multiple alignments for epitope selection, the new RMPs, named A/C-2, A/C-,4, and A/C-7.1, were expressed in E .…”

“…An indirect ELISA assay was performed, with sensitivity and specificity values of 88.89% and 85.54%, respectively. Schreterova et al (2017) [45] conducted a study aimed at developing new RMPs for diagnosing Lyme disease, caused by Borrelia burgdorferi. After using phage display and multiple alignments for epitope selection, the new RMPs, named A/C-2, A/C-,4, and A/C-7.1, were expressed in E. coli (XL1-Blue) cells.…”

Production, characterization, and epitope mapping of monoclonal antibodies of ribosomal protein S3 (rpS3)

“…Epitope characterization can also help elucidate the mechanism of antibody binding (Davidson et al 2015 ) and can enforce intellectual property (patent) protection for the novel antibodies (Deng et al 2018 ; Ledford 2018 ). Experimental epitope mapping data can be incorporated into robust algorithms to facilitate the in silico prediction of B-cell epitopes based on sequence and/or structural data (Schreterova et al 2017 ; Leclair and Lim 2020 ). Epitopes are generally divided into two classes: linear and conformational.…”