Person:
Benito Peña, María Elena

Profile Picture
First Name
María Elena
Last Name
Benito Peña
URI
https://hdl.handle.net/20.500.14352/76070
Affiliation
Universidad Complutense de Madrid
Faculty / Institute
Ciencias Químicas
Department
Química Analítica
Area
Química Analítica
Identifiers
UCM identifierORCIDScopus Author IDDialnet IDGoogle Scholar ID

Filters

2020 - 20213
Reset filters

Settings

Author: Barderas, Rodrigo × Has files: true ×

Search Results

Now showing 1 - 3 of 3
  • Thumbnail Image
    Item
    Recombinant antibodies and their use for food immunoanalysis
    (Analytical & Bioanalytical Chemistry, 2021) Peltomaa, Riikka Johanna; Barderas, Rodrigo; Benito Peña, María Elena; Moreno Bondi, María Cruz
    Antibodies are widely employed as biorecognition elements for the detection of a plethora of compounds including food and environmental contaminants, biomarkers, or illicit drugs. They are also applied in therapeutics for the treatment of several disorders. Recent recommendations from the EU on animal protection and the replacement of animal-derived antibodies by non-animal-derived ones have raised a great controversy in the scientific community. The application of recombinant antibodies is expected to achieve a high growth rate in the years to come thanks to their versatility and beneficial characteristics in comparison to monoclonal and polyclonal antibodies, such as stability in harsh conditions, small size, relatively low production costs, and batch-to-batch reproducibility. This review describes the characteristics, advantages, and disadvantages of recombinant antibodies including antigen-binding fragments (Fab), single-chain fragment variable (scFv), and single-domain antibodies (VHH) and their application in food analysis with especial emphasis on the analysis of biotoxins, antibiotics, pesticides, and foodborne pathogens. Although the wide application of recombinant antibodies has been hampered by a number of challenges, this review demonstrates their potential for the sensitive, selective, and rapid detection of food contaminants.
  • Thumbnail Image
    Item
    Bioluminescent detection of zearalenone using recombinant peptidomimetic Gaussia luciferase fusion protein
    (2020) Peltomaa, Riikka Johanna; Fikacek, Sabrina; Benito Peña, María Elena; Barderas, Rodrigo; Head, Trajen; Deo, Sapna; Daunert, Sylvia; Moreno Bondi, María Cruz
    The development of a bioluminescent immunosensor is reported for the determination of zearalenone (ZEA) based on a peptide mimetic identified by phage display. The peptide mimetic GW, with a peptide sequence GWWGPYGEIELL, was used to create recombinant fusion proteins with the bioluminescent Gaussia luciferase (GLuc) that were directly used as tracers for toxin detection in a competitive immunoassay without the need for secondary antibodies or further labeling. The bioluminescent sensor, based on protein G–coupled magnetic beads for antibody immobilization, enabled determination of ZEA with a detection limit of 4.2 ng/mL (corresponding to 420 μg/kg in food samples) and an IC50 value of 11.0 ng/mL. The sensor performance was evaluated in spiked maize and wheat samples, with recoveries ranging from 87 to 106% (RSD < 20%, n = 3). Finally, the developed method was applied to the analysis of a naturally contaminated reference matrix material and good agreement with the reported concentrations was obtained.
  • Thumbnail Image
    Item
    Identification of high-affinity phage-displayed VH fragments by use of a quartz crystal microbalance with dissipation monitoring
    (Sensors and Actuators: B. Chemical, 2021) Gómez-Arribas, Lidia ; Juste-Dolz, Augusto; Peltomaa, Riikka Johanna; Giménez-Romero, David; Morais, Sergi; Barderas, Rodrigo; Cuadrado, Carmen; Maquieira, Ángel; Benito Peña, María Elena; Moreno Bondi, María Cruz
    Phage display has become a powerful tool for antibody discovery in a wide variety of fields. This technology allows specific binders for a given antigen to be selected from combinatorial libraries. A key step in the process is characterizing and evaluating antibody clones thus selected to reliably identify the best antigen binders. Novel characterization methods can provide essential insight into the binding mechanism and supplement the information obtained with conventional techniques. In this work, we used a quartz crystal microbalance with dissipation monitoring (QCM-D) to determine the kinetic and thermodynamic binding parameters for phage-displayed VH antibody fragments. Phytohemagglutinin (PHA), a legume lectin of analytical interest, was used as a complex model antigen to select specific VH fragments from a phage-displayed library. Eight VH fragments with a unique amino acid sequence were identified as PHA binders by using the well-established enzyme-linked immunosorbent assay (ELISA). QCM-D measurements, structural analysis and principal component analysis (PCA) were used to evaluate the antibody fragments and identify clone clusters with similar binding characteristics and molecular interaction mechanisms. This unprecedented study has enabled the identification of high-affinity phage-displayed VH antibody fragments for PHA, which could be useful for PHA analysis (apparent association constant ranged from 10e8 to 10e10 1/M). In fact, the proposed methodology provides a useful tool for evaluating and characterizing antibody fragments with capabilities beyond those of conventional techniques.