Isolation and purification of recombinant outer surface protein C (rOspC) of Borrelia burgdorferi Sensu Lato

Křupka, Michal; Bĕláková, Jana; Šebestová, Martina; Tuhácková, Jana; Raška, Milan; Vrzal, V.; Weigl, E.

doi:10.5507/bp.2005.036

Cited by 6 publications

(4 citation statements)

References 6 publications

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“…Furthermore, experimental animals and humans have different pattern of epitope recognition too [ 5 , 42 – 46 ]. Nevertheless, given the extensive use of His-tags and their potential use for efficacious affinity purification and oriented anchoring of antigenic proteins such as rOspC onto surface such as liposomes for vaccination purposes [ 14 , 18 , 19 , 47 ] then there is a clear need to demonstrate the potentially pivotal significance to protein immunogenicity should the His-tag be located at N' or C' termini.…”

Section: Discussionmentioning

confidence: 99%

The Position of His-Tag in Recombinant OspC and Application of Various Adjuvants Affects the Intensity and Quality of Specific Antibody Response after Immunization of Experimental Mice

et al. 2016

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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.

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Section: Discussionmentioning

confidence: 99%

The Position of His-Tag in Recombinant OspC and Application of Various Adjuvants Affects the Intensity and Quality of Specific Antibody Response after Immunization of Experimental Mice

et al. 2016

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“…It indicates an advantage of the non-lipidated forms of these antigens even at the cost of lower immunogenicity of these truncated form and the need for addition of adjuvants. Furthermore, lipidated forms of outer surface proteins are problematic for expression in E. coli Krupka et al 2005) in comparison with non-lipidated forms (Krupka et al 2008). …”

Section: Ospa-based Vaccinementioning

confidence: 99%

Prevention of Lyme Disease: Promising Research or Sisyphean Task?

Křupka

Zachová

Weigl

et al. 2011

Arch. Immunol. Ther. Exp.

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Borrelia burgdorferi sensu lato (Spirochaetes) is a group of at least 12 closely related species, some of which are responsible for chronic zoonotic infection that may cause Lyme disease. The only experimentally confirmed vector transmitting Borrelia to mammals is the Ixodes ticks. Borrelia is a highly adapted pathogen that can survive in the host organism in spite of the intense immune responses. Some patients have chronic long-lasting complications despite antibiotic therapy, probably due to adverse effects of the immune responses. A preventive vaccine against this bacterium has not been available due to the relatively broad spectrum and antigenic variability of Borrelia-surface lipoproteins and the different epitope recognition by experimental animals and humans. Although a human vaccine was marketed in the USA, it has been already pulled off the market. In addition, this vaccine was effective only in the USA, where the only pathogenic species is B. burgdorferi sensu stricto. Recent data indicate that a broadly effective vaccine will to be composed of a mixture of several antigens or multiple epitopes.

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“…Recombinant proteins purified by the method of affinity chromatography 18,19 rOspA of B. afzelii, B. burgdorferii origin (rOspA/BA, rOspA/BB) and rOspC of B. garinii origin (rOspC/BG) were tested for immunogenity on the mouse model. 20 Forty inbred BALB/c mice (AnLabLtd.)…”

Section: Introductionmentioning

confidence: 99%

Testing of the Biocan® B inj. ad us. vet. vaccine and development of the new recombinant vaccine against canine borreliosis

Tuhácková¹,

Bĕláková²,

Křupka³

et al. 2005

BIOMED PAP

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Verification of the efficacy of Biocan B inj. ad us. vet. (Bioveta, a.s.) was done by challenge testing. Ticks collected in the nature were used as natural vectors of the infection. Six beagles and two control ones were used in the test. Formation of outer surface protein A specific antibodies (OspA antibodies) and borrelia specific immonoglobulins (IgG) was measured by Western blot and EIA in the sera samples. The tissue samples were used for detection of borreliae by cultivation method and dark field microscopy (DFM). Formation of IgG antibodies and OspA antibodies after vaccination was observed. The maximum titer level of antibodies was reached between 21. and 49. day after vaccination and then slowly decreased. Presence of borreliae was detected only in skin biopsies of non-vaccinated dogs. The post mortem tissue samples showed presence of borreliae in all of the samples of the non-vaccinated dogs. The tissues of the vaccinated dogs were not infected with borreliae, except for two samples of dog with low titer levels of OspA antibodies. The development of the new vaccine is based on preparation of recombinant outer surface proteins (e.g. rOspA and rOspC) of B. afzelii, B. burgdorferi and B. garinii origin. Chosen recombinant proteins were successfully expressed in E. coli. The obtained purified proteins are currently being tested on laboratory BALB/c mice. Formation of specific antibodies against some recombinant proteins has been confirmed. These proteins are suitable candidates for preparation of a vaccine prototype and they will be subsequently used in challenge tests.

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Isolation and purification of recombinant outer surface protein C (rOspC) of Borrelia burgdorferi Sensu Lato

Cited by 6 publications

References 6 publications

The Position of His-Tag in Recombinant OspC and Application of Various Adjuvants Affects the Intensity and Quality of Specific Antibody Response after Immunization of Experimental Mice

The Position of His-Tag in Recombinant OspC and Application of Various Adjuvants Affects the Intensity and Quality of Specific Antibody Response after Immunization of Experimental Mice

Prevention of Lyme Disease: Promising Research or Sisyphean Task?

Testing of the Biocan® B inj. ad us. vet. vaccine and development of the new recombinant vaccine against canine borreliosis

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