Identification of B‐cell epitopes of <i>Borrelia burgdorferi</i> outer surface protein C by screening a phage‐displayed gene fragment library

Pulzová, Lucia; Flachbartova, Zuzana; Bencúrová, Elena; Potocnakova, Lenka; Comor, Lubos; Schreterova, Eva; Bhide, Mangesh

doi:10.1111/1348-0421.12438

Cited by 14 publications

(17 citation statements)

References 33 publications

(47 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The OspC epitopes of B. burgdorferi SKT-2 (AY597037.1) were mapped with the help of phage display in our previous study [9]. Epitopes in other genospecies (B. bavariensis SKT-7.1 GU906888.1 and B. afzelii strain SKT-4 AY518771.1; both strains isolated in Slovakia) were defined with multiple alignment of OspC sequences using SKT-2 as reference.…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

“…The antigen-specific antibodies were affinity purified, as described previously [9]. Briefly, pre-swollen CNBrSepharose 4B beads (Pharmacia Fine Chemicals) were mixed with purified his-tagged OspA or OspC proteins of SKT-2, SKT-4 and SKT-7.1 (previously produced in the author's laboratory [9,13]). Total IgGs (previously purified in the author's laboratory [9]) were incubated with beads for 3 hrs at room temperature, and antigen-specific antibodies eluted from the beads with 3.6M MgCl 2 .…”

Section: Methodsmentioning

confidence: 99%

“…Besides OspA, outer surface protein OspC is a promising candidate for the construction of an LD diagnostic marker due to its high immunologic potential and its early expression during LD infection [8]. Six OspC B-cell epitopes were identified recently using phage display [9].…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

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

Schreterova

Bhide

Potocnakova

et al. 2017

Ann Agric Environ Med.

Self Cite

View full text Add to dashboard Cite

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.

show abstract

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

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

Schreterova

Bhide

Potocnakova

et al. 2017

Ann Agric Environ Med.

Self Cite

View full text Add to dashboard Cite

show abstract

“…SKT-2 OspC via a 12-mer phage-displayed peptide library (45). The library was selected by panning with human LD patient antibodies that had been affinity purified using the same OspC.…”

Section: Figmentioning

confidence: 99%

Identification of Surface Epitopes Associated with Protection against Highly Immune-Evasive VlsE-Expressing Lyme Disease Spirochetes

et al. 2018

View full text Add to dashboard Cite

The tick-borne pathogen is responsible for approximately 300,000 Lyme disease (LD) cases per year in the United States. Recent increases in the number of LD cases, in addition to the spread of the tick vector and a lack of a vaccine, highlight an urgent need for designing and developing an efficacious LD vaccine. Identification of protective epitopes that could be used to develop a second-generation (subunit) vaccine is therefore imperative. Despite the antigenicity of several lipoproteins and integral outer membrane proteins (OMPs) on the surface, the spirochetes successfully evade antibodies primarily due to the VlsE-mediated antigenic variation. VlsE is thought to sterically block antibody access to protective epitopes of However, it is highly unlikely that VlsE shields the entire surface epitome. Thus, identification of subdominant epitope targets that induce protection when they are made dominant is necessary to generate an efficacious vaccine. Toward the identification, we repeatedly immunized immunocompetent mice with live-attenuated VlsE-deleted and then challenged the animals with the VlsE-expressing (host-adapted) wild type. Passive immunization and Western blotting data suggested that the protection of 50% of repeatedly immunized animals against the highly immune-evasive was antibody mediated. Comparison of serum antibody repertoires identified in protected and nonprotected animals permitted the identification of several putative epitopes significantly associated with the protection. Most linear putative epitopes were conserved between the main pathogenic genospecies and found within known subdominant regions of OMPs. Currently, we are performing immunization studies to test whether the identified protection-associated epitopes are protective for mice.

show abstract

Epitope mapping of Borrelia burgdorferi OspC protein in homodimeric fold

Norek

Janda

2017

Protein Science

View full text Add to dashboard Cite

In current work, we used recombinant OspC protein derived from B. afzelii strain BRZ31 in the native homodimeric fold for mice immunization and following selection process to produce three mouse monoclonal antibodies able to bind to variable parts of up to five different OspC proteins. Applying the combination of mass spectrometry assisted epitope mapping and affinity based theoretical prediction we have localized regions responsible for antigen-antibody interactions and approximate epitopes' amino acid composition. Two mAbs (3F4 and 2A9) binds to linear epitopes located in previously described immunogenic regions in the exposed part of OspC protein. The third mAb (2D1) recognises highly conserved discontinuous epitope close to the ligand binding domain 1.

show abstract

Identification of B‐cell epitopes of Borrelia burgdorferi outer surface protein C by screening a phage‐displayed gene fragment library

Cited by 14 publications

References 33 publications

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

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

Identification of Surface Epitopes Associated with Protection against Highly Immune-Evasive VlsE-Expressing Lyme Disease Spirochetes

Epitope mapping of Borrelia burgdorferi OspC protein in homodimeric fold

Contact Info

Product

Resources

About