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.

• Klíčová slova
• Borrelia burgdorferi, OspC, epitope mapping, immunoprecipitation, monoclonal antibody, outer surface protein C, protein alignment,
• MeSH
• antigeny bakteriální chemie   genetika   imunologie   MeSH
• Borrelia burgdorferi chemie   genetika   imunologie   MeSH
• mapování epitopu * MeSH
• multimerizace proteinu * MeSH
• myši inbrední BALB C MeSH
• myší monoklonální protilátky chemie   imunologie   MeSH
• myši MeSH
• proteiny vnější bakteriální membrány chemie   genetika   imunologie   MeSH
• sbalování proteinů MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigeny bakteriální MeSH
• myší monoklonální protilátky MeSH
• OspC protein MeSH Prohlížeč
• proteiny vnější bakteriální membrány MeSH

Lyme disease, Borrelia burgdorferi-caused infection, if not recognized and appropriately treated by antibiotics, may lead to chronic complications, thus stressing the need for protective vaccine development. The immune protection is mediated by phagocytic cells and by Borrelia-specific complement-activating antibodies, associated with the Th1 immune response. Surface antigen OspC is involved in Borrelia spreading through the host body. Previously we reported that recombinant histidine tagged (His-tag) OspC (rOspC) could be attached onto liposome surfaces by metallochelation. Here we report that levels of OspC-specific antibodies vary substantially depending upon whether rOspC possesses an N' or C' terminal His-tag. This is the case in mice immunized: (a) with rOspC proteoliposomes containing adjuvants MPLA or non-pyrogenic MDP analogue MT06; (b) with free rOspC and Montanide PET GEL A; (c) with free rOspC and alum; or (d) with adjuvant-free rOspC. Stronger responses are noted with all N'-terminal His-tag rOspC formulations. OspC-specific Th1-type antibodies predominate post-immunization with rOspC proteoliposomes formulated with MPLA or MT06 adjuvants. Further analyses confirmed that the structural features of soluble N' and C' terminal His-tag rOspC and respective rOspC proteoliposomes are similar including their thermal stabilities at physiological temperatures. On the other hand, a change in the position of the rOspC His-tag from N' to C' terminal appears to affect substantially the immunogenicity of rOspC arguably due to steric hindrance of OspC epitopes by the C' terminal His-tag itself and not due to differences in overall conformations induced by changes in the His-tag position in rOspC variants.

• MeSH
• adjuvancia imunologická * MeSH
• antigeny bakteriální aplikace a dávkování   chemie   imunologie   MeSH
• Borrelia burgdorferi imunologie   MeSH
• ELISA MeSH
• imunizace MeSH
• lymeská nemoc imunologie   MeSH
• modely u zvířat MeSH
• myši MeSH
• proteiny vnější bakteriální membrány aplikace a dávkování   chemie   imunologie   MeSH
• proteolipidy MeSH
• protilátky bakteriální imunologie   MeSH
• rekombinantní fúzní proteiny aplikace a dávkování   chemie   imunologie   izolace a purifikace   MeSH
• sekundární struktura proteinů MeSH
• specificita protilátek imunologie   MeSH
• stabilita proteinů MeSH
• tvorba protilátek imunologie   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• adjuvancia imunologická * MeSH
• antigeny bakteriální MeSH
• OspC protein MeSH Prohlížeč
• proteiny vnější bakteriální membrány MeSH
• proteolipidy MeSH
• proteoliposomes MeSH Prohlížeč
• protilátky bakteriální MeSH
• rekombinantní fúzní proteiny MeSH