Darién García

Centro de Ingeniería Genética y Biotecnología

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Karem Cobas

Centro de Ingeniería Genética y Biotecnología

Olivia Niebla

Centro de Ingeniería Genética y Biotecnología

Evelín Caballero

Centro de Ingeniería Genética y Biotecnología

Maité Delgado

National University of San Marcos

Gretel Sardiñas

Centro de Ingeniería Genética y Biotecnología

Daniel Yero

Universitat Autònoma de Barcelona

Yasser Perera

Centro de Ingeniería Genética y Biotecnología

Sonia González

Centro de Ingeniería Genética y Biotecnología

Rolando Pajón

Moderna Therapeutics (United States)

Lázaro Betancourt

Lund University

Cooperative Institutions

Centro de Ingeniería Genética y Biotecnología

National Center for Genetic Engineering and Biotechnology

National University of Quilmes

Instituto de Salud Carlos III

ORCID

Biogipuzkoa Health Research Institute

Instituto de Medicina Tropical “Pedro Kourí”

Finlay Institute

Biomedical Research Networking Center on Neurodegenerative Diseases

Centro de Investigación Biomédica en Red

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Epitope mapping of anti-human transferrin monoclonal antibodies: potential uses for transferrin–transferrin receptor interaction studies

Journal of Molecular Recognition (2008)

Yasser Perera Darién García Osmany Guirola Vivian Huerta Y Beovides García

Abstract Human transferrin (hTf) is an 80 kDa glycoprotein involved in iron transport from the absorption sites to the sites of storage and utilization. Additionally, transferrin also plays a relevant role as a bacteriostatic agent preventing uncontrolled bacterial growth in the host. In this work we describe a well‐characterized Mabs panel in terms of precise epitope localization and estimate affinity for the two major hTf isoforms. We found at least four antigenic regions in the hTf molecule, narrowed down the interacting antigen residues within three of such regions, and located them on a molecular model of hTf. Two of the antigenic regions partially overlap with previously described transferrin‐binding sites for both human receptor (antigenic region I: containing amino acid residues from Asp‐69 to Asn‐76 at the N‐lobe) and bacterial receptors from two pathogenic species (antigenic region III: amino acid residues from Leu‐665 to Ser‐672 at the C‐lobe). Hence, such monoclonal antibodies (Mabs) could be used as an additional tool for conformational studies and/or the characterization of the interaction between hTf and both types of receptor molecules. Copyright © 2008 John Wiley & Sons, Ltd.

Transferrin receptor

Epitope mapping

10.1002/jmr.878

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Citations (2)

Two mucosal–parenteral schedules to coadminister a multiantigenic formulation against HIV-1 in Balb/c mice

International Immunopharmacology (2012)

Enrique V. Iglesias Daymir García Gabriel Márquez Yayrí C. Prieto Jorge Sánchez

Priming (agriculture)

HIV Antigens

10.1016/j.intimp.2011.12.026

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Citations (7)

Predicted proteins of Neisseria meningitidis as potential vaccine candidates: from in silico analyses to experimental corroboration

Biotecnología aplicada (2012)

Darién García Daniel Yero Olivia Niebla Karem Cobas Yasser Perera

Neisseria meningitidis serogroup B infections are a serious health threat to the world that cannot be prevented by vaccination. Here, we report an analysis of the MC58 Neisseria meningitidis genome aimed at the identification of new potential vaccine candidates. 'Hypothetical' and 'conserved hypothetical' annotated genes, together with those with putative functions related to the cell envelope, were subjected to extensive sequence similarity searches, as well as motif, cellular location, and domain analyses complemented with manual curation. As a result, a set of 35 unchar- acterized ORFs, predicted to encode for surface exposed or virulence related proteins, was identified. The candidates were subdivided in three categories: 1) predicted outer membrane proteins (OMPs) unique of the Neisseria genus; 2) conserved OMPs from various genus and 3) proteins homologous to known OMPs or to proteins previously found to be immunogenic in animal models. Two of the final candidates, nmb1126 and nmb0181, were cloned and expressed in Escherichia coli. The resulting products were purified by Metal Chelating Chromatography and used to immunize mice. The recombinant proteins were capable of inducing antibodies against the native antigen in preparations of a panel of three strains and displayed bactericidal activity against the homologous strains.

ORFS

Neisseria

Reverse vaccinology

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Bicistronic expression plasmid for the rapid production of recombinant fused proteins in Escherichia coli

Biotechnology and Applied Biochemistry (2006)

Daniel Yero Rolando Pajón Olivia Niebla Gretel Sardiñas Isbel Vivar

In the post‐genomic era, every aspect of the production of proteins must be accelerated. In this way, several vectors are currently exploited for rapid production of recombinant proteins in Escherichia coli . N‐terminal fusions to the first 47 amino acids of the LpdA (dihydrolipoamide dehydrogenase A) protein of Neisseria meningitidis have been shown to increase the expression of recombinant proteins. Consequently, we have constructed a modified N‐terminal LpdA fusion vector, introducing the blue/white colony selection by exploiting a bicistronic gene organization. In the new vector, the sequence encoding the first 47 amino acids of meningococcal LpdA and the α‐peptide sequence of β‐galactosidase were connected via a ribosome‐binding site, and two MCSs (multiple cloning sites) were located surrounding the latter, allowing efficient cloning by colour selection of recombinants. The vector was also improved with the addition of a C‐terminal polyhistidine tag, and an EKS (enterokinase recognition sequence) immediately after the LpdA fusion sequence. The new plasmid was employed in the expression and purification of six different bacterial polypeptides. One of these recombinant proteins, P6 protein from Haemophilus influenzae , was used as a model and its N‐terminal fusion sequence was totally removed from the recombinant version after incubation with the enterokinase protease, while the polyhistidine tail successfully allowed the purification of the unfused protein from the protease reaction. Two completely new neisserial vaccine candidates, NMB0088 and NMB1126 proteins, were cloned, expressed and purified using this system. To our knowledge, this constitutes the first report of the cloning and expression of these proteins in E. coli .

10.1042/ba20050170

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Citations (10)

Identification of new meningococcal serogroup B surface antigens through a systematic analysis of neisserial genomes

Vaccine (2009)

Rolando Pajón Daniel Yero Olivia Niebla Yanet Climent Gretel Sardiñas

Neisseria

Meningococcal vaccine

10.1016/j.vaccine.2009.09.128

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Citations (21)

20919. ICTUS ISQUÉMICO POR EMBOLISMO TUMORAL COMO DEBUT DE UN ADENOCARCINOMA DE PULMÓN

Neurology Perspectives (2024)

Ignacio Gómez C. Jauregui Larrañaga G. Núñez Manjarres A. Val-Carreres Castellote Sónia González

10.1016/s2667-0496(24)00443-5

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Citations (0)

Evaluación económica del tratamiento con Heberprot P® para las úlceras del pie diabético en el Centro de Atención al Diabético La Habana, 2012–2014

PharmacoEconomics Spanish Research Articles (2017)

Manuel Collazo Herrera Irma Sosa Lorenzo Lisbeth Fernández González Darién García B Miranda

10.1007/s40277-017-0071-4

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Caracterización de proteínas presentes en proteoliposoma de N. meningitidis. Estudios de consistencia y reproducibilidad entre lotes de VA-MENGOC-BC® mediante técnicas proteómicas

Biotecnología aplicada (2007)

Lila Castellanos Lázaro Betancourt Gretel Sardiñas Yasser Perera Olivia Niebla

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Optimization of a Therapeutic Vaccine Candidate by Studying Routes, Immunization Schedules and Antigen Doses in HBsAg-positive Transgenic Mice

Euroasian Journal of Hepato-Gastroenterology (2013)

Heidy Trujillo A Blanco Darién García Freya Freyre J. Aguiar

Hepatitis B core antigens (HBcAg) and hepatitis B surface antigens (HBsAg) are the main structural antigens of hepatitis B virus (HBV). Both antigens are potent immunogens for experimental animals as well as in acutely infected patients. A novel formulation based on the combination of HBsAg and HBcAg has been developed as a therapeutic vaccine candidate, aimed at inducing an immune response capable of controlling the infection. An immunization schedule was conducted to evaluate the immunogenicity of this formulation after simultaneous immunization by the intranasal and parenteral routes using different schedules and doses. Humoral and cellular immune responses generated in blood and spleen were evaluated by engyme-linked immunosorbent assay (ELISA) and enzyme-liked immunospot (ELISPOT) assays respectively. A first experiment evaluated two groups of mice simultaneously immunized by intranasal (IN) and subcutaneous (SC) routes, one including alum by SC route and, in the other, the formulation was injected without adjuvant. As a result, alum adjuvant did not increase the immunogenicity under the studied conditions. In fact, the group without alum induced the most potent immune response. The immune response was enhanced by combining IN and SC immunization compared to the SC route alone. In a second experiment, mice were immunized by different mucosal routes at the same time, and compared to the simultaneously (IN/SC) immunized groups. It was demonstrated that there is no improvement on the resulting immune response by using multiple routes of immunizations simultaneously; however, the increase of the antigen dose induced a superior immune response. Interestingly, the increase of antigen dose only by SC route did not favor the resulting immunogenicity. In conclusion, the use of HBsAg transgenic mice has proven useful to optimize the formulation, avoiding the unnecessary use of alum as adjuvant as well as provided information of the role of different mucosal immunization routes and antigen dose on the resulting immune response. How to cite this article: Trujillo H, Blanco A, García D, Freyre F, Aguiar J, Lobaina Y, Aguilar JC. Optimization of a Therapeutic Vaccine Candidate by Studying Routes, Immunization Schedules and Antigen Doses in HBsAg-positive Transgenic Mice. Euroasian J Hepato-Gastroenterol 2014;4(2):70-78.

ELISPOT

HBcAg

Alum

10.5005/jp-journals-10018-1105

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Citations (3)

Th2-Th1 shift with the multiantigenic formulation TERAVAC-HIV-1 in Balb/c mice

Immunology Letters (2012)

Darién García Ingrid B. Rodriguez Yaimín Santisteban Gabriel Márquez Yanet Terrero

BALB/c

10.1016/j.imlet.2012.11.007

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Citations (4)