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Development and comparison of the first electrochemical bioplatforms for determining anti-centromere B antibodies (CENPB-Abs) developed in magnetic microbead-assisted or integrated formats using His-tag chemistry.

An electrochemical biosensing platform for serum autoantibodies (AAbs) detection is reported in this work, exploiting for the first time six Alzheimer's disease (AD)-specific phage-derived and frameshift aberrant HaloTag peptides as receptors, immobilized on magnetic microbeads (MBs) surface and captured on disposable electrodes to perform amperometric detection. Operational analytical characteristics and clinical diagnostic ability of the bioplatform were probed in optimized key experimental conditions by analysing serum AAbs of AD patients and healthy subjects. The value of 100 % obtained for AUC, sensitivity, and selectivity from the all peptides combined ROC curve, indicate full AD-diagnostic capability of the methodology, which was further implemented, as proof of concept, in a POC multiplexing platform to detect the signature in a single test over clinically actionable times (1 h 15 min), opening great promise for the type of diagnosis and AD patients’ monitoring follow-up currently pursued. The financial support of RTI2018-096135-B−I00 (Spanish Ministerio de Ciencia, Innovación y Universidades) and PID2019-103899RB−I00 (Spanish Ministerio de Ciencia e Innovación), Research Projects, PI17CIII/00045 and PI20CIII/00019 grants from the AES-ISCIII program and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349) are gratefully acknowledged. A. Montero-Calle and P. San Segundo-Acosta acknowledge the support of the FPU predoctoral contracts by the Spanish Ministerio de Educación, Cultura y Deporte. A. Valverde acknowledges a predoctoral contract from Complutense University of Madrid. B. Arévalo acknowledges a predoctoral contract from the Spanish Ministerio de Economía y Competitividad (PRE2019-087596). G.S-F. is a recipient of a predoctoral contract (grant number 1193818N) supported by The Flanders Research Foundation (FWO). Sí

This work reports the first amperometric biosensor for the simultaneous determination of the single or total content of the most relevant human immunoglobulin isotypes (hIgs) of anti-dsDNA antibodies, dsDNA-hIgG, dsDNA-hIgM, dsDNA-hIgA and dsDNA-three hIgs, which are considered relevant biomarkers in prevalent autoimmune diseases such as systemic lupus erythematosus (SLE) as well as of interest in neurodegenerative diseases such as Alzheimer's disease (AD). The bioplatform involves the use of neutravidin-functionalized magnetic microparticles (NA-MBs) modified with a laboratory-prepared biotinylated human double-stranded DNA (b-dsDNA) for the efficient capture of specific autoantibodies that are enzymatically labeled with horseradish peroxidase (HRP) enzyme using specific secondary antibodies for each isotype or a mixture of secondary antibodies for the total content of the three isotypes. Transduction was performed by amperometry (-0.20 V vs. the Ag pseudo-reference electrode) using the H2O2/hydroquinone (HQ) system after trapping the resulting magnetic bioconjugates on each of the four working electrodes of a disposable quadruple transduction platform (SP4CEs). The bioplatform demonstrated attractive operational characteristics for clinical application and was employed to determine the individual or total hIgs classes in serum from healthy individuals and from patients diagnosed with SLE and AD. The target concentrations in AD patients are provided for the first time in this work. In addition, the results for SLE patients and control individuals agree with those obtained by applying ELISA tests as well as with the clinical ranges reported by other authors, using individual detection methodologies restricted to centralized settings or clinical laboratories. The financial support of PID2019-103899RB-I00 and PID2021-122457OB-I00 (Spanish Ministerio de Ciencia e Innovación) Research Projects, PI17CIII/00045 and PI20CIII/00019 Grants from the AESISCIII Program co-founded by FEDER funds and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349) are gratefully acknowledged. B.A. acknowledges predoctoral contracts from the Spanish Ministerio de Ciencia, Innovación y Universidades (PRE2019-087596). M.G-A. acknowledges the postdoctoral contract Margarita Salas for the requalification of the Spanish University System. A.M-C. was supported by a FPU predoctoral contract supported by the Spanish Ministerio de Educación, Cultura y Deporte. Sí

A dual immunosensor is reported for the simultaneous determination of two important immunity-related cytokines: BAFF (B cell activation factor) and APRIL (a proliferation-induced signal). Sandwich-type immunoassays with specific antibodies (cAbs) and a strategy for signal amplification based on labelling the detection antibodies (dAbs) with binary MoS2/MWCNTs nanostructures and using horseradish peroxidase (HRP) were implemented. Amperometric detection was carried out at screen-printed dual carbon electrodes (SPdCEs) through the hydroquinone HQ/H2O2 system. The developed dual immunosensor provided limit of detection (LOD) of 0.08 and 0.06 ng mL-1 for BAFF and APRIL, respectively, and proved to be useful for the determination of both cytokines in cancer cell lysates and serum samples from patients diagnosed with autoimmune diseases and cancer. The obtained results agreed with those found using ELISA methodologies. Open Access funding provided thanks to the CRUE-CSIC agreement with Springer Nature. Financial support was provided by Spanish Ministerio de Ciencia, Innovacion y Universidades (RTI2018-096,135-B-I00), Spanish Ministerio de Ciencia e Innovación (PID2019-103899RB-I00) Research Project, and the TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349). Financial support of the PI17CIII/00045 and PI20CIII/00019 grants from the AES-ISCIII program was also provided to R. B. A predoctoral contract from the Spanish Ministerio de Ciencia, Innovacion y Universidades (PRE2019-087,596) was provided to B. Arévalo. A FPU predoctoral contract supported by the Spanish Ministerio de Educación, Cultura y Deporte was provided to A. M.-C. G. S.-F. is recipient of a predoctoral contract (grant number 1193818 N) supported by The Flanders Research Foundation (FWO). Sí

This work reports the first electrochemical bioplatform for the determination of soy traces in food. The bioplatform involves sandwich-type immunoassays using specific antibodies for β-conglycinin and glycinin, which are the main allergenic soy proteins, and carboxylic acid-modified magnetic microbeads. Amperometric detection at −0.20 V (vs. an Ag pseudo-reference electrode) was performed using single or dual screen-printed carbon electrodes and the H2O2/hydroquinone (HQ) system. The measured variation in the cathodic current was directly proportional to the concentration of target allergenic proteins. The developed bioplatforms exhibit a good selectivity and sensitivity providing limits of detection (LOD) values of 0.03 and 0.02 ng mL−1 for β-conglycinin and glycinin, respectively. The determination of both proteins can be carried out in only 1.5 h. The electrochemical bioplatforms allow their accurate determinations (with results statistically comparable to those provided by ELISA methodologies) in raw cookie dough and baked cookies enriched with soy flour. The results obtained confirm, in a pioneering way with electrochemical biosensors, the possibility of discriminating samples incurred with as little as 0.0005 ppm of a food allergen in model cookie extracts., The financial support of PID2019-103899RB-I00 to S.C., (Spanish Ministerio de Ciencia e Innovación) Research Project, and TRANSNANOAVANSENS-CM Program from the Comunidad de Madrid (Grant S2018/NMT-4349) to S.C. are gratefully acknowledged. S.B. acknowledges a Juan de la Cierva Incorporación contract (Ref.: IJCI-2017-31345) from the Spanish Ministerio de Economía, Industria y Competitividad.

A dual immunosensor is reported for the simultaneous determination of two important immunity-related cytokines: BAFF (B cell activation factor) and APRIL (a proliferation-induced signal). Sandwich-type immunoassays with specific antibodies (cAbs) and a strategy for signal amplification based on labelling the detection antibodies (dAbs) with binary MoS

This work reports the first electroanalytical bioplatform to date for the determination of antibodies against aquaporin-4 (AQP4-Abs), whose serum level is considered as relevant biomarker for certain autoimmune diseases. The bioplatform relies on the use of magnetic microparticles modified with the biotinylated protein for the capture of specific antibodies. The captured IgGs are enzymatically labelled with a secondary antibody conjugated to the horseradish peroxidase (HRP) enzyme. Amperometric transduction is performed using the H

This work reports the first amperometric immunosensor for the simultaneous determination of four fertility-related hormones in saliva: progesterone (P4), luteinizing hormone (LH), estradiol (E2), and prolactin (PRL). The immune platform involves direct competitive (P4 and E2), and sandwich (LH and PRL) assays implemented onto functionalized magnetic microbeads (MBs). The amperometric transduction was performed upon placing the MBs-immunoconjugates onto each of the four working electrodes of the SPCE array (SP4CEs) and applying a detection potential of -0.20 V (vs. Ag pseudo-reference electrode) using the H2O2/hydroquinone (HQ) system. The achieved analytical and operational characteristics of the developed multiplexed immunoplatform showed a sensitivity that allows the determination of these hormones in saliva, and an adequate selectivity to analyse complex clinical samples. The bioplatform was employed for the determination of the set of four hormones in human saliva samples collected from individuals with different hormonal profiles. The results obtained using a conventional potentiostat were compared with those provided employing a novel low-cost custom-designed and field-portable quadruple potentiostat. Similar results were found which also agreed with those obtained by applying ELISA methods for the determination of single hormones.

An electrochemical immunosensor for the fast, sensitive, and selective determination of salivary prolactin (PRL) is reported in this work. The developed methodology uses a sandwich-type immunoassay implemented on magnetic microparticles (MBs) functionalized with neutravidin (NA-MBs) where the capture antibody was immobilized. The resulting immunocomplexes were placed onto a screen-printed carbon electrode (SPCE) and the target hormone quantification was made by amperometry at − 0.20 V (vs. Ag pseudo-reference electrode) using the HQ/H2O2 system as electron transfer mediator and enzyme substrate. The developed immunosensor allows the determination of PRL in a linear range between 1.1 and 100 ng mL−1 (R2 = 0.996) with a limit of detection (LOD) value of 0.32 ng mL−1 PRL, and shows an excellent selectivity. The whole assay protocol takes 46 min. The immunosensor was able to determine PRL in saliva with results in agreement with those provided by ELISA method. In addition, the developed immunosensor was applied to the determination of PRL in breast milk.

Matrix metalloproteinase 9 (MMP-9) is a zinc-dependent endopeptidase that promotes angiogenesis, tumor growth, metastasis and cell invasion through the degradation of extracellular matrix. This work reports a magnetic microbeads (MBs)-based sandwich immunoassay for the amperometric determination of MMP-9 at screen-printed carbon electrodes (SPCEs). The suitable capture antibody (cAb) is immobilized onto carboxylic MBs to selectively capture the antigen which is sandwiched with a biotinylated detector antibody (biotin-dAb) further conjugated with a commercial streptavidin-horseradish peroxidase (Strep-HRP) polymer. This immunoplatform provides great analytical characteristics in terms of selectivity and sensitivity, achieving a LOD value of 2.4 pg mL−1 for standards in buffered solutions. Although this value is similar to those reported for some other approaches described so far, the method described here is simpler involving a single 30 min incubation step which makes it ideal for automation or implementation in POC devices. Moreover, the method was assayed for the accurate determination of endogenous MMP-9 in both cancer cell lysates and serum samples of patients diagnosed with different subtypes of breast cancer (BC) after a simple dilution. The results obtained show that the disposable and affordable immunoplatform developed is able not only to discriminate BC patients from healthy individuals but also to do it for the worst outcome triple negative (TNBC) subtype.

4′-Substituted derivatives of 2,2′:6′,2′′-terpyridine with N-containing heteroaromatic substituents, such as pyridyl groups, might be able to coordinate metal centres through the extra N-donor atom, in addition to the chelating terpyridine N atoms. The incorporation of these peripheral N-donor sites would also allow for the diversification of the types of noncovalent interactions present, such as hydrogen bonding and π-π stacking. The title compound, C24H16N4, consists of a 2,2′:6′,2′′-terpyridine nucleus (tpy), with a pendant isoquinoline group (isq) bound at the central pyridine (py) ring. The tpy nucleus deviates slightly from planarity, with interplanar angles between the lateral and central py rings in the range 2.24(7)-7.90(7)°, while the isq group is rotated significantly [by 46.57(6)°] out of this planar scheme, associated with a short Htpy⋯Hisq contact of 2.32Å. There are no strong noncovalent interactions in the structure, the main ones being of the π-π and C - H⋯π types, giving rise to columnar arrays along [001], further linked by C - H⋯N hydrogen bonds into a three-dimensional supramolecular structure. An Atoms In Molecules (AIM) analysis of the noncovalent interactions provided illuminating results, and while confirming the bonding character for all those interactions unquestionable from a geometrical point of view, it also provided answers for some cases where geometric parameters are not informative, in particular, the short Htpy⋯Hisq contact of 2.32Å to which AIM ascribed an attractive character. Fil: Granifo, Juan. Universidad de La Frontera; Chile Fil: Arévalo, Beatriz. Universidad de La Frontera; Chile Fil: Gaviño, Rubén. Universidad Nacional Autónoma de México; México Fil: Suarez, Sebastian. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Ciudad Universitaria. Instituto de Química, Física de los Materiales, Medioambiente y Energía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales. Instituto de Química, Física de los Materiales, Medioambiente y Energía; Argentina. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; Argentina Fil: Baggio, Ricardo Fortunato. Comisión Nacional de Energía Atómica. Centro Atómico Constituyentes; Argentina

This work reports the first amperometric biosensor involving the use of neutravidin-functionalized magnetic microbeads (NA-MBs) modified with a biotinylated-anti-dsDNA (b-dsDNA) as efficient magnetic microcarriers to selectively capture anti-dsDNA autoantibodies (IgG, IgA and IgM AAbs) present in the sera of patients with rheumatoid arthritis (RA). Subsequently, the attached anti-dsDNA AAbs are detected with a mixture of conventional HRP-labeled secondary antibodies (HRP-anti-human IgG/IgM/IgA mixture). The biorecognition event is monitored by amperometric transduction using the hydroquinone (HQ)/H2O2 system upon capturing the modified MBs on the surface of screen-printed carbon electrodes (SPCEs). The developed bioplatform exhibits a linear calibration plot ranging from 1 to 200 IU mL−1 with a LOD of 0.3 IU mL−1 for anti-dsDNA AAbs standards. In addition, the biosensor allows performing the determination of the anti-dsDNA AAbs levels directly in 100-times diluted serum samples from patients diagnosed with RA and in just 75 min. The obtained results are in agreement with those provided by an ELISA kit and allow discrimination between positive and negative samples.

En este artículo se muestran algunos resultados de investigación que hacen evidentes las dificultades de los estudiantes para poner en práctica el conocimiento adquirido, es decir, el poco desarrollo de competencias científicas en el aula y particularmente la del uso del conocimiento científico. Poniendo de manifiesto la importancia de desarrollar estrategias como la resolución de problemas, donde el estudiante sea un agente activo en la construcción de conocimiento.

This work describes a new electroanalytical device for the simultaneous and reliable determination of two fertility relevant hormones (luteinizing hormone, LH, and progesterone, P4) in saliva. The device is constructed using a custom designed and field-portable potentiostat where dual disposable immunosensing platform are connected. The immunosensors are based on sandwich-type and competitive immunoassays implemented onto magnetic microbeads (MBs) functionalized with Neutravidin and Protein G for the determination of LH and P4, respectively. Amperometric detection performed at −0.20 V vs the Ag pseudo-reference electrode using the H2O2/hydroquinone (HQ) system was employed as the transduction technique after placing the MBs with immunocomplexes for each target hormone on the appropriate working electrode of screen-printed dual carbon electrodes (SPdCEs). The method exhibits high sensitivity and selectivity for the target hormones providing detection limits of 1.7 pg mL−1 and 0.10 mIU mL−1 for P4 and LH, respectively, with a 1 h test time. The applicability of the method was confirmed by determining both hormones in saliva samples from different volunteers providing results comparable to those obtained using amperometric transduction with a commercial potentiostat and with ELISA methodologies involving the same immunoreagents for each target hormone.