Please login first
Florencio M. Ubeira  - - - 
Top co-authors See all
Humberto González-Díaz

215 shared publications

Department of Organic Chemistry II , University of the Basque Country UPV/EHU , 48940 Leioa , Spain

Grace Patlewicz

128 shared publications

National Center for Computational Toxicology, Office of Research and Development (ORD), U.S. Environmental Protection Agency (EPA), Research Triangle Park, North Carolina, USA

Esther Lete

124 shared publications

Department of Organic Chemistry II , University of the Basque Country UPV/EHU , 48940 Leioa , Spain

Paulino Gomez-Puertas

119 shared publications

Centro de Biologia Molecular “Severo Ochoa” (CSIC‐UAM) Campus Universidad Autonoma de Madrid MadridSpain

Cristian-Robert Munteanu

112 shared publications

Department of Computation, Faculty of Computer Science, A Coruña, Spain

116
Publications
0
Reads
0
Downloads
237
Citations
Publication Record
Distribution of Articles published per year 
(1993 - 2019)
Total number of journals
published in
 
30
 
Publications See all
Article 0 Reads 0 Citations Antibody responses to chimeric peptides derived from parasite antigens in mice and other animal species R.A. Orbegozo-Medina, V. Martínez-Sernández, I. Folgueira, M... Published: 01 February 2019
Molecular Immunology, doi: 10.1016/j.molimm.2018.11.019
DOI See at publisher website
Article 0 Reads 0 Citations In-plate recapturing of a dual-tagged recombinant Fasciola antigen (FhLAP) by a monoclonal antibody (US9) prevents non-s... Ricardo A. Orbegozo-Medina, Victoria Martínez-Sernández, Mar... Published: 01 February 2019
PLOS ONE, doi: 10.1371/journal.pone.0211035
DOI See at publisher website ABS Show/hide abstract
Recombinant proteins expressed in E. coli are frequently purified by immobilized metal affinity chromatography (IMAC). By means of this technique, tagged proteins containing a polyhistidine sequence can be obtained up to 95% pure in a single step, but some host proteins also bind with great affinity to metal ions and contaminate the sample. A way to overcome this problem is to include a second tag that is recognized by a preexistent monoclonal antibody (mAb) in the gene encoding the target protein, allowing further purification. With this strategy, the recombinant protein can be directly used as target in capture ELISA using plates sensitized with the corresponding mAb. As a proof of concept, in this study we engineered a Trichinella-derived tag (MTFSVPIS, recognized by mAb US9) into a His-tagged recombinant Fasciola antigen (rFhLAP) to make a new chimeric recombinant protein (rUS9-FhLAP), and tested its specificity in capture and indirect ELISAs with sera from sheep and cattle. FhLAP was selected since it was previously reported to be immunogenic in ruminants and is expressed in soluble form in E. coli, which anticipates a higher contamination by host proteins than proteins expressed in inclusion bodies. Our results showed that a large number of sera from non-infected ruminants (mainly cattle) reacted in indirect ELISA with rUS9-FhLAP after single-step purification by IMAC, but that this reactivity disappeared testing the same antigen in capture ELISA with mAb US9. These results demonstrate that the 6XHis and US9 tags can be combined when double purification of recombinant proteins is required.
Article 0 Reads 0 Citations In-plate recapturing of a dual-tagged recombinant Fasciola antigen (FhLAP) by a monoclonal antibody (US9) prevents non-s... Ricardo A. Orbegozo-Medina, Victoria Martínez-Sernández, Mar... Published: 01 February 2019
PLOS ONE,
PubMed View at PubMed ABS Show/hide abstract
Recombinant proteins expressed in E. coli are frequently purified by immobilized metal affinity chromatography (IMAC). By means of this technique, tagged proteins containing a polyhistidine sequence can be obtained up to 95% pure in a single step, but some host proteins also bind with great affinity to metal ions and contaminate the sample. A way to overcome this problem is to include a second tag that is recognized by a preexistent monoclonal antibody (mAb) in the gene encoding the target protein, allowing further purification. With this strategy, the recombinant protein can be directly used as target in capture ELISA using plates sensitized with the corresponding mAb. As a proof of concept, in this study we engineered a Trichinella-derived tag (MTFSVPIS, recognized by mAb US9) into a His-tagged recombinant Fasciola antigen (rFhLAP) to make a new chimeric recombinant protein (rUS9-FhLAP), and tested its specificity in capture and indirect ELISAs with sera from sheep and cattle. FhLAP was selected since it was previously reported to be immunogenic in ruminants and is expressed in soluble form in E. coli, which anticipates a higher contamination by host proteins than proteins expressed in inclusion bodies. Our results showed that a large number of sera from non-infected ruminants (mainly cattle) reacted in indirect ELISA with rUS9-FhLAP after single-step purification by IMAC, but that this reactivity disappeared testing the same antigen in capture ELISA with mAb US9. These results demonstrate that the 6XHis and US9 tags can be combined when double purification of recombinant proteins is required.
Article 4 Reads 1 Citation Comparison of recombinant cathepsins L1, L2, and L5 as ELISA targets for serodiagnosis of bovine and ovine fascioliasis Victoria Martínez-Sernández, María J. Perteguer, Ana Hernand... Published: 21 March 2018
Parasitology Research, doi: 10.1007/s00436-018-5809-7
DOI See at publisher website
Article 0 Reads 0 Citations Vaccination of sheep with Quil-A® adjuvant expands the antibody repertoire to the Fasciola MF6p/FhHDM-1 antigen and admi... Ricardo A. Orbegozo-Medina, Victoria Martínez-Sernández, Mar... Published: 08 March 2018
Vaccine, doi: 10.1016/j.vaccine.2018.02.115
DOI See at publisher website PubMed View at PubMed
Article 5 Reads 0 Citations Fasciola spp: Mapping of the MF6 epitope and antigenic analysis of the MF6p/HDM family of heme-binding proteins Victoria Martínez-Sernández, María J. Perteguer, Mercedes Me... Published: 21 November 2017
PLOS ONE, doi: 10.1371/journal.pone.0188520
DOI See at publisher website PubMed View at PubMed ABS Show/hide abstract
MF6p/FhHDM-1 is a small cationic heme-binding protein which is recognized by the monoclonal antibody (mAb) MF6, and abundantly present in parenchymal cells and secreted antigens of Fasciola hepatica. Orthologs of this protein (MF6p/HDMs) also exist in other causal agents of important foodborne trematodiasis, such as Clonorchis sinensis, Opisthorchis viverrini and Paragonimus westermani. Considering that MF6p/FhHDM-1 is relevant for heme homeostasis in Fasciola and was reported to have immunomodulatory properties, this protein is expected to be a useful target for vaccination. Thus, in this study we mapped the epitope recognized by mAb MF6 and evaluated its antigenicity in sheep. The sequence of the MF6p/FhHDM-1 ortholog from F. gigantica (MF6p/FgHDM-1) was also reported. By means of ELISA inhibitions with overlapping synthetic peptides, we determined that the epitope recognized by mAb MF6 is located within the C-terminal moiety of MF6p/FhHDM-1, which is the most conserved region of MF6p/HDMs. By immunoblotting analysis of parasite extracts and ELISA inhibitions with synthetic peptides we also determined that mAb MF6 reacted with the same intensity with F. hepatica and F. gigantica, and in decreasing order of intensity with C. sinensis, O.viverrini and P. westermani orthologs. On the contrary, mAb MF6 showed no reactivity against Dicrocoelium dendriticum and Schistosoma mansoni. The study of the recognition of peptides covering different regions of MF6p/FhHDM-1 by sera from immunized sheep revealed that the C-terminal moiety is the most antigenic, thus being of potential interest for vaccination. We also demonstrated that the production of antibodies to MF6p/FhHDM-1 in sheep infected by F. hepatica occurs relatively early and follows the same pattern as those produced against L-cathepsins.
Top