NEW MONOCLONAL ANTIBODIES FOR IN SITU DETECTION OF MYCOTOXINS IN FRUITS, CEREALS AND PROCESSED PRODUCTS USING IMMUNOCHEMICAL METHODS (Q3189118): Difference between revisions
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Revision as of 15:48, 11 October 2021
Project Q3189118 in Spain
| Language | Label | Description | Also known as |
|---|---|---|---|
| English | NEW MONOCLONAL ANTIBODIES FOR IN SITU DETECTION OF MYCOTOXINS IN FRUITS, CEREALS AND PROCESSED PRODUCTS USING IMMUNOCHEMICAL METHODS |
Project Q3189118 in Spain |
Statements
84,700.0 Euro
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169,400.0 Euro
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50.0 percent
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1 January 2019
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31 December 2021
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AGENCIA CONSEJO SUPERIOR DE INVESTIGACIONES CIENTIFICAS
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39°30'14.11"N, 0°26'31.56"W
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46190
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ENTRE LAS SUSTANCIAS POTENCIALMENTE PRESENTES EN ALIMENTOS Y PIENSOS, LAS MICOTOXINAS REPRESENTAN UN SERIO PROBLEMA PARA LA SALUD HUMANA Y ANIMAL, OCASIONANDO IMPORTANTES PERDIDAS ECONOMICAS AL SECTOR AGROALIMENTARIO. PARA ESTE GRUPO DE CONTAMINANTES, LOS METODOS BASADOS EN LA UTILIZACION DE ANTICUERPOS GOZAN DE UNA AMPLIA ACEPTACION A NIVEL MUNDIAL COMO TECNICAS DE SCREENING. NO OBSTANTE, HEMOS DETECTADO QUE, PARA ALGUNAS MICOTOXINAS COMO LA PATULINA Y EL ALTERNARIOL, NO EXISTEN METODOS INMUNOQUIMICOS COMERCIALMENTE DISPONIBLES, Y EN OTROS CASOS, COMO LA ZEARALENONA, LAS ESTRATEGIAS SEGUIDAS PARA LA OBTENCION DE INMUNORREACTIVOS SON MANIFIESTAMENTE MEJORABLES. ANTE ESTA SITUACION, HEMOS CONSIDERADO COMO UN OBJETIVO DE GRAN TRASCENDENCIA CIENTIFICO-TECNOLOGICA EL DESARROLLO DE METODOS INMUNOQUIMICOS PARA LA DETERMINACION SELECTIVA DE ESTAS TRES IMPORTANTES BIOTOXINAS EN AQUELLAS MATRICES ALIMENTARIAS DONDE SU PRESENCIA PUDIERA CONSTITUIR UN RIESGO._x000D_ COMO PASO PREVIO PARA DESARROLLAR SISTEMAS BIOANALITICOS ESPECIFICOS PARA CADA UNA DE LAS TRES MICOTOXINAS CONTEMPLADAS, SERA NECESARIO GENERAR LOS INMUNORREACTIVOS (HAPTENOS, CONJUGADOS Y ANTICUERPOS), ASI COMO CARACTERIZARLOS Y COMBINARLOS ADECUADAMENTE PARA LOGRAR LAS PRESTACIONES EXIGIBLES A CUALQUIER METODOLOGIA ANALITICA._x000D_ LOS OBJETIVOS PARTICULARES DE ESTE SUBPROYECTO SE PUEDEN RESUMIR DEL SIGUIENTE MODO:_x000D_ - PRODUCCION DE BIORRECEPTORES. SE GENERARA UNA COLECCION DE ANTICUERPOS POLICLONALES Y MONOCLONALES CAPACES DE RECONOCER CON ELEVADA AFINIDAD Y ESPECIFICAMENTE CADA UNA DE LAS TRES MICOTOXINAS (ALTERNARIOL, ZEARALENONA Y PATULINA)._x000D_ - CARACTERIZACION DE ANTICUERPOS Y BIOCONJUGADOS. SE DETERMINARA LA AFINIDAD Y ESPECIFICIDAD DE LOS BIORRECEPTORES EN DIFERENTES FORMATOS DE ENSAYO Y UTILIZANDO BIOCONJUGADOS HOMOLOGOS Y HETEROLOGOS._x000D_ - EVALUACION DE CONSTRUCTOS BASADOS EN NANOTUBOS PARA OBTENER ANTICUERPOS MONOCLONALES. SE ESTUDIARA LA IDONEIDAD PARA PREPARAR ANTICUERPOS MONOCLONALES DE CONSTRUCTOS BASADOS EN NANOTUBOS DE CARBONO, CON O SIN PROTEINA, SOBRE LOS QUE SE INMOVILIZARA DE MANERA COVALENTE O SE ADSORBERAN LOS HAPTENOS INMUNIZANTES. _x000D_ - DESARROLLO DE INMUNOENSAYOS TIPO ELISA. SE ESTUDIARAN DIFERENTES FORMATOS DE INMUNOENSAYO PARA LOS QUE SE DETERMINARA LA INFLUENCIA DE LOS PARAMETROS FISICOQUIMICOS MAS RELEVANTES Y SE OPTIMIZARA LAS CONDICIONES DE ENSAYO. _x000D_ - PRODUCCION DE COLUMNAS DE INMUNOAFINIDAD. SE ESTUDIARA LA INMOVILIZACION DE LOS ANTICUERPOS Y LA CAPACIDAD DE LAS COLUMNAS PARA LA CAPTURA DE LAS MICOTOXINAS._x000D_ - DESARROLLO DE ENSAYOS INMUNOCROMATOGRAFICOS. LOS PARAMETROS DE ESTUDIO MAS DESTACADOS DE ESTE METODO CONSISTEN EN LA INMOVILIZACION DE INMUNORREACTIVOS SOBRE UNA MEMBRANA, LA PREPARACION DE BIOCONJUGADOS SOBRE PARTICULAS DE ORO COLOIDAL, Y LA OPTIMIZACION DE LAS CONDICIONES DE ENSAYO._x000D_ - VALIDACION DE LOS METODOS ANALITICOS DESARROLLADOS. SE ESTABLECERA UN CONJUNTO DE PROTOCOLOS PARA LA APLICACION DE LOS INMUNOENSAYOS AL ANALISIS DE CADA MICOTOXINA EN DIVERSAS MATRICES DE ALIMENTOS O PIENSOS. (Spanish)
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AMONG THE CONTAMINANTS POTENTIALLY PRESENT IN FOOD AND FEED, MYCOTOXINS REPRESENT A MAJOR PROBLEM FOR HUMAN AND ANIMAL HEALTH, CAUSING IMPORTANT ECONOMIC COSTS TO THE AGRO-FOOD INDUSTRY. FOR THIS GROUP OF CONTAMINANTS, ANTIBODY-BASED METHODS ARE WIDELY IMPLEMENTED IN LABORATORIES WORLDWIDE AS SCREENING TECHNIQUES. HOWEVER, WE HAVE DETECTED THAT FOR SOME MYCOTOXINS, SUCH AS PATULIN AND ALTERNARIOL, THERE ARE NO COMMERCIALLY AVAILABLE IMMUNOCHEMICAL METHODS, AND IN OTHER CASES, SUCH AS ZEARALENONE, THE STRATEGIES THAT HAVE BEEN FOLLOWED TO OBTAIN IMMUNOREAGENTS CAN BE IMPROVED. IN VIEW OF THIS SITUATION, WE HAVE CONSIDERED THAT THE DEVELOPMENT OF IMMUNOCHEMICAL METHODS FOR THE SELECTIVE DETERMINATION OF THESE THREE RELEVANT BIOTOXINS, IN THOSE FOOD AND FEED MATRICES WHERE THEY COULD CONSTITUTE A RISK, IS AN OBJECTIVE OF GREAT SCIENTIFIC AND TECHNOLOGICAL IMPORTANCE._x000D_ THE GROUP THAT PARTICIPATES IN THIS SUBPROJECT HAS A LARGE RESEARCH EXPERIENCE IN OBTAINING MOLECULAR BIORECEPTORS AND IN DEVELOPING DIFFERENT IMMUNOCHEMICAL METHODS, PARTICULARLY APPLIED TO THE ANALYSIS OF SMALL ORGANIC MOLECULES IN FOOD, AS REFLECTED IN THE PATENTS DEPOSITED, LICENSED AND UNDER EXPLOITATION, AS WELL AS IN THE NUMEROUS PUBLICATIONS IN THE MOST PRESTIGIOUS JOURNALS OF AREAS SUCH AS THE ANALYTICAL CHEMISTRY AND THE FOOD CHEMISTRY._x000D_ AS A PRELIMINARY STEP TO DEVELOP SPECIFIC BIOANALYTICAL SYSTEMS FOR EACH OF THE THREE MYCOTOXINS SELECTED FOR THIS PROJECT, IT WILL BE NECESSARY TO GENERATE THE IMMUNOREAGENTS (HAPTENS, CONJUGATES, AND ANTIBODIES), AS WELL AS TO CHARACTERIZE AND COMBINE THEM ADEQUATELY IN ORDER TO ACHIEVE THE ADEQUATE PERFORMANCE THAT IS REQUIRED TO ANY ANALYTICAL METHODOLOGY._x000D_ THE PARTICULAR OBJECTIVES OF THIS SUBPROJECT CAN BE SUMMARIZED AS FOLLOWS:_x000D_ - PRODUCTION OF BIORECEPTORS. A COLLECTION OF HIGH-AFFINITY AND SPECIFIC POLYCLONAL AND MONOCLONAL ANTIBODIES CAPABLE OF RECOGNIZING EACH OF THE THREE MYCOTOXINS (ALTERNARIOL, ZEARALENONE AND PATULIN) WILL BE GENERATED._x000D_ - CHARACTERIZATION OF ANTIBODIES AND BIOCONJUGATES. THE AFFINITY AND SPECIFICITY OF THE OBTAINED BIORECEPTORS WILL BE DETERMINED IN DIFFERENT ASSAY FORMATS AND USING HOMOLOGOUS AND HETEROLOGOUS BIOCONJUGATES._x000D_ - EVALUATION OF NANOTUBE-BASED CONSTRUCTS TO OBTAIN MONOCLONAL ANTIBODIES. THE SUITABILITY OF CONSTRUCTS, WITH OR WITHOUT PROTEIN, BASED ON CARBON NANOTUBES TO PREPARE MONOCLONAL ANTIBODIES WILL BE STUDIED WHEN THE IMMUNIZING HAPTEN HAS BEEN COVALENTLY IMMOBILIZED OR SIMPLY ADSORBED ON THE PARTICLES._x000D_ - DEVELOPMENT OF ELISA-TYPE IMMUNOASSAYS. DIFFERENT IMMUNOASSAY FORMATS WILL BE STUDIED FOR WHICH THE INFLUENCE OF THE MOST RELEVANT PHYSICOCHEMICAL PARAMETERS WILL BE DETERMINED, AND THE ASSAY CONDITIONS WILL BE OPTIMIZED._x000D_ - PRODUCTION OF IMMUNOAFFINITY COLUMNS. THE IMMOBILIZATION PROCESS OF ANTIBODIES AND THE ABILITY OF THE OBTAINED COLUMNS TO CAPTURE TOXINS WILL BE STUDIED._x000D_ - DEVELOPMENT OF IMMUNOCHROMATOGRAPHIC ASSAYS. THE MOST IMPORTANT PARAMETERS THAT WILL HAVE TO BE STUDIED CONSIST IN THE IMMOBILIZATION OF IMMUNOREAGENTS ON THE MEMBRANE, THE PREPARATION OF BIOCONJUGATES USING COLLOIDAL GOLD PARTICLES, AND THE OPTIMIZATION OF THE ASSAY CONDITIONS._x000D_ - VALIDATION OF THE DEVELOPED ANALYTICAL METHODS. A SET OF PROTOCOLS WILL BE ESTABLISHED FOR THE APPLICATION OF IMMUNOASSAYS TO THE ANALYSIS OF EACH MYCOTOXIN IN VARIOUS FOOD OR FEED MATRICES. (English)
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Paterna
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Identifiers
RTI2018-096121-B-C21
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