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Epitope mapping of Borrelia burgdorferi OspC protein in homodimeric fold.

ABSTRACT: 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.

SUBMITTER: Norek A

PROVIDER: S-EPMC5368064 | biostudies-literature | 2017 Apr

REPOSITORIES: biostudies-literature

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Epitope mapping of Borrelia burgdorferi OspC protein in homodimeric fold.

Norek Adam A Janda Lubomír L

Protein science : a publication of the Protein Society 20170211 4

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' am ...[more]

PMID: 28142214

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Protein Microarray Analysis of Antibody Responses of Experimentally Infected Dogs to Borrelia burgdorferi Antigens OspC

Project description:Lyme borreliosis (LB) is a tick-borne infection caused by Borrelia burgdorferi. Dogs are at high risk of exposure to ticks and tick-borne pathogens, including B. burgdorferi. Immunodiagnostic assays are usually based on whole-cell preparations of B. burgdorferi as substrate and, consequently, interpretation of results is confounded by antibody cross-reactivity between borrelial antigens and other bacterial species, as well as the anti-LB vaccination status of the dog. For this study, we examined sera from 33 dogs that were experimentally infected with B. burgdorferi through tick bite. These sera were compared with sera from uninfected dogs in their reactivities to 72 different recombinant B. burgdorferi antigens and 24 OspC protein types on a protein microarray. Amongst antigens frequently recognized by infected dogs were several known to be immunogens for humans, such as Decorin-binding protein A (BBA25), BBA64, fibronectin-binding protein (BBK32), VlsE, Erp and Bdr, CRASP proteins, OspC proteins and some flagellar antigens. Of special interest were the novel antigens BBB14 and BB0844, both hypothetical lipoproteins about which very little is currently known, and that were frequently and strongly recognized by infected dog sera. The antibody response of B. burgdorferi-infected dogs presents both similarities and differences from human counterparts, and both can be important for improvement of canine LB diagnosis and vaccine development. Antibody profiling was performed on sera from dogs experimentally-infected with B. burgdorferi and unexposed controls against antigens of B. burgdorferi. Thirty-three serum samples from experimental infections, and 5 unexposed controls were probed on a protein microarray displaying 24 OspC proteins of B. burgdorferi .

2013-08-22 | E-GEOD-50075 | biostudies-arrayexpress

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