Person: Vivanco Martínez, Fernando
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First Name
Fernando
Last Name
Vivanco Martínez
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Area
Bioquímica y Biología Molecular
Identifiers
32 results
Search Results
Now showing 1 - 10 of 32
Item Novel liquid chromatography–mass spectrometry method for sensitive determination of the mustard allergen Sin a 1 in food(Food Chemistry, 2015) Maria Posada-Ayala; Álvarez Llamas, Gloria; Aroa S. Maroto; Xavier Maes; Esther Muñoz-Garcia; Villalba Díaz, María Teresa; Rodríguez García, Rosalía; Marina Perez-Gordo; Vivanco Martínez, Fernando; Pastor Vargas, Carlos; Javier Cuesta-HerranzMustard is a condiment added to a variety of foodstuffs and a frequent cause of food allergy. A new strategy for the detection of mustard allergen in food products is presented. The methodology is based on liquid chromatography analysis coupled to mass spectrometry. Mustard allergen Sin a 1 was purified from yellow mustard seeds. Sin a 1 was detected with a total of five peptides showing a linear response (lowest LOD was 5ng). Sin a 1 was detected in mustard sauces and salty biscuit (19±3mg/kg) where mustard content is not specified. Sin a 1, used as an internal standard, allowed quantification of this mustard allergen in foods. A novel LC/MS/MS SRM-based method has been developed to detect and quantify the presence of mustard. This method could help to detect mustard allergen Sin a 1 in processed foods and protect mustard-allergic consumers.Item Identification of thaumatin‐like protein and aspartyl protease as new major allergens in lettuce (Lactuca sativa).(Molecular nutrition and food research, 2013) Muñoz‐García, Esther; Luengo‐Sánchez, Olga; Haroun‐Díaz, Elisa; Sanz Maroto, Aroa; Palacín, Arancha; Díaz –Perales, Araceli; de las Heras Gozalo, Manuel; Labrador‐Horrillo, Moisés; Vivanco Martínez, Fernando; Cuesta‐Herranz, Javier; Pastor Vargas, CarlosScope: Today, about 2-8% of the population of Western countries exhibits some type of food allergy whose impact ranges from localized symptoms confined to the oral mucosa to severe anaphylactic reactions. Consumed worldwide, lettuce is a Compositae family vegetable that can elicit allergic reactions. To date, however, only one lipid transfer protein has been described in allergic reaction to lettuce. The aim of this study was to identify potential new allergens involved in lettuce allergy. Methods and results: Sera from 42 Spanish lettuce-allergic patients were obtained from patients recruited at the outpatient clinic. IgE-binding proteins were detected by SDS-PAGE and immunoblotting. Molecular characterization of IgE-binding bands was performed by MS. Thaumatin was purified using the Agilent 3100 OFFGEL system. The IgE-binding bands recognized in the sera of more than 50% of patients were identified as lipid transfer protein (9 kDa), a thaumatin-like protein (26 kDa), and an aspartyl protease (35 and 45 kDa). ELISA inhibition studies were performed to confirm the IgE reactivity of the purified allergen. Conclusion: Two new major lettuce allergens-a thaumatin-like protein and an aspartyl protease-have been identified and characterized. These allergens may be used to improve both diagnosis and treatment of lettuce-allergic patients.Item Occupational rhinitis due to inhalation of chicken meat protein.(J Investig Allergol Clin Immunol, 2014) Lobera Labairu, T; González Mahave, I; Del Pozo Gil, M D; Venturini Díaz, M; Blasco Sarramián, A; Pastor Vargas, Carlos; Vivanco Martínez, Fernando; Bartolomé Zavala, BorjaItem Detection of major food allergens in amniotic fluid: initial allergenic encounter during pregnancy(Pediatr Allergy Immunol, 2016) Pastor Vargas, Carlos; Aroa S. Maroto; Araceli Díaz‐Perales; Villalba Díaz, María Teresa; Vanesa Esteban; Marta Ruiz‐Ramos; Marta Rodriguez de Alba; Vivanco Martínez, Fernando; Javier Cuesta‐HerranzBackground: Ingestion of food allergens present in maternal milk during breastfeeding has been hypothesized as a gateway to sensitization to food; however, this process could develop during pregnancy, as the maternal-fetal interface develops a Th2- and Treg-mediated environment to protect the fetus. We hypothesized that in these surroundings, unborn children are exposed to food allergens contained in the mother's diet, possibly giving rise to first sensitization. Methods: The presence of allergens in utero was studied by analyzing amniotic fluid (AF) samples in two different stages of pregnancy: at 15-20 weeks and after delivery at term. An antibody microarray was developed to test for the most common food allergens. The array detects the presence of ten allergens from milk, fruit, egg, fish, nuts, and wheat. Results: AF from 20 pregnant women was collected: eight after delivery at term and 12 from women who underwent diagnostic amniocentesis between weeks 15 and 20 of gestation. The presence of allergens was detected in all samples. Samples from amniocentesis had a higher allergen concentration than samples after delivery at term. Conclusions: We demonstrated the presence of intact major food allergens in AF samples. This early contact could explain subsequent sensitization to foods never eaten before.Item Sensitive detection of major food allergens in breast milk: first gateway for allergenic contact during breastfeeding(Allergy, 2015) Pastor Vargas, Carlos; Maroto, Aroa; Díaz‐Perales, Araceli; Villalba Díaz, María Teresa; Casillas Diaz, Natalia; Vivanco Martínez, Fernando; Cuesta‐Herranz, JavierFood allergy is recognized as a major public health issue, especially in early childhood. It has been hypothesized that early sensitization to food allergens maybe due to their ingestion as components dissolved in the milk during the breastfeeding, explaining reaction to a food, which has never been taken before. Thus, the aim of this work has been to detect the presence of the food allergens in breast milk by microarray technology. We produced a homemade microarray with antibodies produced against major food allergens. The antibody microarray was incubated with breast milk from 14 women collected from Fundación Jiménez Díaz Hospital. In this way, we demonstrated the presence of major foods allergens in breast milk. The analysis of allergens presented in breast milk could be a useful tool in allergy prevention and could provide us a key data on the role of this feeding in tolerance induction or sensitization in children.Item Novel liquid chromatography–mass spectrometry method for sensitive determination of the mustard allergen Sin a 1 in food(Food Chemistry, 2015) Posada-Ayala, Maria; Álvarez Llamas, Gloria; Maroto, Aroa; Maes, Xavier; Muñoz-Garcia, Esther; Villalba Díaz, María Teresa; Rodríguez García, Rosalía; Perez-Gordo, Marina; Vivanco Martínez, Fernando; Pastor Vargas, Carlos; Cuesta-Herranz, JavierMustard is a condiment added to a variety of foodstuffs and a frequent cause of food allergy. A new strategy for the detection of mustard allergen in food products is presented. The methodology is based on liquid chromatography analysis coupled to mass spectrometry. Mustard allergen Sin a 1 was purified from yellow mustard seeds. Sin a 1 was detected with a total of five peptides showing a linear response (lowest LOD was 5ng). Sin a 1 was detected in mustard sauces and salty biscuit (19±3mg/kg) where mustard content is not specified. Sin a 1, used as an internal standard, allowed quantification of this mustard allergen in foods. A novel LC/MS/MS SRM-based method has been developed to detect and quantify the presence of mustard. This method could help to detect mustard allergen Sin a 1 in processed foods and protect mustard-allergic consumers.Item Watermelon Profilin: Characterization of a Major Allergen as a Model for Plant-Derived Food Profilins(International Archives of Allergy Immunology, 2010) Cases, Bárbara ; Pastor Vargas, Carlos; Gil Dones, Félix; Perez-Gordo, Marina; Maroto, Aroa; las Heras, Manuel de las; Vivanco Martínez, Fernando; Cuesta-Herranz, JavierBackground: Plant profilins have been reported as minor allergens. They are a well-known pan-allergen family responsible for cross-reactivity between plant-derived foods and pollens. Watermelon profilin has been reported to be a major allergen in watermelon (Citrullus lanatus).The aim of this study was to characterize recombinant watermelon profilin, confirming its reactivity for diagnostic purposes and the development of immunotherapy. Methods: Native profilin was purified from watermelon extract by affinity chromatography using poly-L-proline. Recombinant His-tagged profilin was produced in Pichia pastoris yeast using pPICZαA vector and purified by metal chelate affinity chromatography. ELISA and immunoblot were carried out with sera from 17 watermelon-allergic patients. Biological activity was tested by the basophil activation test. Results: Native profilin and recombinant profilin were purified and identified by mass spectrometry. Both show similar IgE reactivity in vitro and are biologically active. Conclusions: Similarities were found in the IgE-binding patterns and biological activity of recombinant profilin and native profilin. Recombinant profilin may be a powerful tool for specific diagnosis.Item Targeting antigens to an invariant epitope of the MHC Class II DR molecule potentiates the immune response to subunit vaccines(Virus Research, 2011) Pérez-Filgueira, Mariano; Barderas, María G.; Alonso, Covadonga; José M, Escribano; Gil Dones, Félix; Pastor Vargas, Carlos; Vivanco Martínez, FernandoRecombinant subunit and peptidic vaccines in general present a reduced immunogenicity in vaccinated individuals with respect to the whole pathogen from which they derived. The generation of strong immune responses to these vaccines requires the use of potent adjuvants, high antigen doses and repetitive vaccinations. In this report, we document the enhanced antibody response obtained against two recombinant subunit vaccines by means of targeting to antigen-presenting cells by a recombinant single chain antibody. This antibody, named APCH1, recognizes an epitope of MHC Class II DR molecule preserved in different animal species, including humans. We showed that vaccinal antigens translationally fused to APCH1 antibody and produced by recombinant baculoviruses in insect larvae (Trichoplusia ni), elicited an increased antibody response in comparison with the same antigens alone or fused to a carrier molecule. These results suggest that targeting of antigens to this invariant MHC Class II epitope has immunopotentiating effects that could circumvent the reduced potency of peptidic or subunit vaccines, opening the possibility of widespread application of APCH1 as a new adjuvant antibody of general use.Item Angler fish is not an alternative in allergy to multiple fish species(ANNALS OF ALLERGY ASTHMA & IMMUNOLOGY, 2014) Julián Azofra; Aroa Sanz Maroto; Vivanco Martínez, Fernando; Pastor Vargas, CarlosFish allergy is one of the most frequent food allergies. 1 Its prevalence is highest is those regions where consumption is greatest. Spain has one of the highest fish intakes per inhabitant and, as a result, fish allergy is a serious health problem.Item Modification of the Secretion Pattern of Proteases, Inflammatory Mediators, and Extracellular Matrix Proteins by Human Aortic Valve is Key in Severe Aortic Stenosis(Molecular and cellular proteomics, 2013) Álvarez Llamas, Gloria; Martín Rojas, Tatiana; Cuesta, Fernando De La; Calvo, Enrique; Gil Dones, Félix; Dardé, Veronica ; López Almodovar, Luis ; Padial, Luis ; Lopez, Juan Antonio; Vivanco Martínez, Fernando; Barderas, MariaOne of the major challenges in cardiovascular medicine is to identify candidate biomarker proteins. Secretome analysis is particularly relevant in this search as it focuses on a subset of proteins released by a cell or tissue under certain conditions. The sample can be considered as a plasma subproteome and it provides a more direct approximation to the in vivo situation. Degenerative aortic stenosis is the most common worldwide cause of valve replacement. Using a proteomic analysis of the secretome from aortic stenosis valves we could identify candidate markers related to this pathology, which may facilitate early diagnosis and treatment. For this purpose, we have designed a method to validate the origin of secreted proteins, demonstrating their synthesis and release by the tissue and ruling out blood origin. The nLC-MS/MS analysis showed the labeling of 61 proteins, 82% of which incorporated the label in only one group. Western blot and selective reaction monitoring differential analysis, revealed a notable role of the extracellular matrix. Variation in particular proteins such as PEDF, cystatin and clusterin emphasizes the link between aortic stenosis and atherosclerosis. In particular, certain proteins variation in secretome levels correlates well, not only with label incorporation trend (only labeled in aortic stenosis group) but, more importantly, with alterations found in plasma from an independent cohort of samples, pointing to specific candidate markers to follow up in diagnosis, prognosis, and therapeutic intervention.