Boosting the sensitivity of in vitro b-lactam allergy diagnostic tests

[EN] The synthesis of structurally new haptens and the development of suitable antigens are essential for boosting the sensitivity of drug allergy diagnostic testing. Unprecedented structural antigens for benzylpenicillin and amoxicillin are characterised and evaluated in a cohort of 70 subjects with a turnkey solution based on consumer electronics.This work was funded by the H2020 program (project COBIOPHAD, grant agreement No. 688448), an initiative of the Photonics Public Private Partnership; Agencia Estatal de Investigacion (CTQ2016-75749-R, FEDER) and program UPV-La FE 2019 (PI05 VALBIOAL). E. P. M. holds a FPU doctoral fellowship from Ministerio de Educación, Cultura y Deporte.Peña-Mendizábal, E.; Morais, S.; Maquieira Catala, Á. (2020). Boosting the sensitivity of in vitro b-lactam allergy diagnostic tests. Chemical Communications. 56(80):11973-11976. https://doi.org/10.1039/D0CC04903DS11973119765680Van Boeckel, T. P., Gandra, S., Ashok, A., Caudron, Q., Grenfell, B. T., Levin, S. A., & Laxminarayan, R. (2014). Global antibiotic consumption 2000 to 2010: an analysis of national pharmaceutical sales data. The Lancet Infectious Diseases, 14(8), 742-750. doi:10.1016/s1473-3099(14)70780-7Khan, D. A., & Solensky, R. (2010). Drug allergy. Journal of Allergy and Clinical Immunology, 125(2), S126-S137.e1. doi:10.1016/j.jaci.2009.10.028Blanca, M., Romano, A., Torres, M. J., Férnandez, J., Mayorga, C., Rodriguez, J., … Atanasković-Marković, M. (2009). Update on the evaluation of hypersensitivity reactions to betalactams. Allergy, 64(2), 183-193. doi:10.1111/j.1398-9995.2008.01924.xWorld Health Organization, WHO Report on Surveillance of Antibiotic Consumption: 2016–2018 Early implementation, Geneva, 2018Blanca-Lopez, N., Jimenez-Rodriguez, T. W., Somoza, M. L., Gomez, E., Al-Ahmad, M., Perez-Sala, D., & Blanca, M. (2019). Allergic reactions to penicillins and cephalosporins: diagnosis, assessment of cross-reactivity and management. Expert Review of Clinical Immunology, 15(7), 707-721. doi:10.1080/1744666x.2019.1619548Chang, C., Mahmood, M. M., Teuber, S. S., & Gershwin, M. E. (2011). Overview of Penicillin Allergy. Clinical Reviews in Allergy & Immunology, 43(1-2), 84-97. doi:10.1007/s12016-011-8279-6Baldo, B. A., & Pham, N. H. (2002). Immunoglobulin E binding determinants on β-lactam drugs. Current Opinion in Allergy and Clinical Immunology, 2(4), 297-300. doi:10.1097/00130832-200208000-00002Weltzien, H. U., & Padovan, E. (1998). Molecular Features of Penicillin Allergy. Journal of Investigative Dermatology, 110(3), 203-206. doi:10.1046/j.1523-1747.1998.00122.xBALDO. (1999). Penicillins and cephalosporins as allergens - structural aspects of recognition and cross-reactions. Clinical & Experimental Allergy, 29(6), 744-749. doi:10.1046/j.1365-2222.1999.00575.xZhao, Z., Baldo, B. A., & Rimmer, J. (2002). β-Lactam allergenic determinants: fine structural recognition of a cross-reacting determinant on benzylpenicillin and cephalothin. Clinical & Experimental Allergy, 32(11), 1644-1650. doi:10.1046/j.1365-2222.2002.01492.xZhao, Z., Baldo, B. A., Baumgart, K. W., & Mallon, D. F. J. (2001). Fine structural recognition specificities of IgE antibodies distinguishing amoxicilloyl and amoxicillanyl determinants in allergic subjects. Journal of Molecular Recognition, 14(5), 300-307. doi:10.1002/jmr.541Peng, J., Kong, D., Liu, L., Song, S., Kuang, H., & Xu, C. (2015). Determination of quinoxaline antibiotics in fish feed by enzyme-linked immunosorbent assay using a monoclonal antibody. Analytical Methods, 7(12), 5204-5209. doi:10.1039/c5ay00953gPeng, D., Ye, S., Wang, Y., Chen, D., Tao, Y., Huang, L., … Yuan, Z. (2011). Development and Validation of an Indirect Competitive Enzyme-Linked Immunosorbent Assay for the Screening of Tylosin and Tilmicosin in Muscle, Liver, Milk, Honey and Eggs. Journal of Agricultural and Food Chemistry, 60(1), 44-51. doi:10.1021/jf2037449Brun, E. M., Garcés-García, M., Puchades, R., & Maquieira, Á. (2004). Enzyme-linked immunosorbent assay for the organophosphorus insecticide fenthion. Influence of hapten structure. Journal of Immunological Methods, 295(1-2), 21-35. doi:10.1016/j.jim.2004.08.014Kim, Y. J., Cho, Y. A., Lee, H.-S., Lee, Y. T., Gee, S. J., & Hammock, B. D. (2003). Synthesis of haptens for immunoassay of organophosphorus pesticides and effect of heterology in hapten spacer arm length on immunoassay sensitivity. Analytica Chimica Acta, 475(1-2), 85-96. doi:10.1016/s0003-2670(02)01037-1Montañez, M. I., Mayorga, C., Torres, M. J., Ariza, A., Blanca, M., & Perez-Inestrosa, E. (2011). Synthetic Approach to Gain Insight into Antigenic Determinants of Cephalosporins: In Vitro Studies of Chemical Structure−IgE Molecular Recognition Relationships. Chemical Research in Toxicology, 24(5), 706-717. doi:10.1021/tx100446gPastor-Navarro, N., Morais, S., Maquieira, Á., & Puchades, R. (2007). Synthesis of haptens and development of a sensitive immunoassay for tetracycline residues. Analytica Chimica Acta, 594(2), 211-218. doi:10.1016/j.aca.2007.05.045Chen, Z.-J., Fu, H.-J., Luo, L., Sun, Y.-M., Yang, J.-Y., Zeng, D.-P., … Xu, Z.-L. (2017). Development of competitive indirect ELISAs with a flexible working range for the simple quantification of melatonin in medicinal foods. Analytical Methods, 9(10), 1617-1626. doi:10.1039/c6ay03380fPastor-Navarro, N., Maquieira, Á., & Puchades, R. (2009). Review on immunoanalytical determination of tetracycline and sulfonamide residues in edible products. Analytical and Bioanalytical Chemistry, 395(4), 907-920. doi:10.1007/s00216-009-2901-ySingh, K. V., Kaur, J., Varshney, G. C., Raje, M., & Suri, C. R. (2003). Synthesis and Characterization of Hapten−Protein Conjugates for Antibody Production against Small Molecules. Bioconjugate Chemistry, 15(1), 168-173. doi:10.1021/bc034158vMas, S., Badran, A. A., Juárez, M.-J., Fernández de Rojas, D. H., Morais, S., & Maquieira, Á. (2020). Highly sensitive optoelectrical biosensor for multiplex allergy diagnosis. Biosensors and Bioelectronics, 166, 112438. doi:10.1016/j.bios.2020.112438Vultaggio, A., Matucci, A., Virgili, G., Rossi, O., Filì, L., Parronchi, P., … Maggi, E. (2009). Influence of total serum IgE levels on thein vitrodetection of β-lactams-specific IgE antibodies. Clinical & Experimental Allergy, 39(6), 838-844. doi:10.1111/j.1365-2222.2009.03219.

Multiparametric Highly Sensitive Chemiluminescence Immunoassayfor Quantification of b-Lactam-Specific Immunoglobulin E

This document is the Accepted Manuscript version of a Published Work that appeared in final form in Analytical Chemistry, copyright © American Chemical Society after peer review and technical editing by the publisher. To access the final edited and published work see https://doi.org/10.1021/acs.analchem.0c03020[EN] beta-lactams (BLCs) are the most widely used antibiotics and consequently the most common cause of drug allergy in the world. The diagnosis of drug allergy is complex and represents a serious challenge that includes a wide variety of methods. In vitro tests are based on the immunological determination of allergen-specific IgE, but the tests in the market lack the required sensitivity and specificity. In addition, the large sample volume, long incubation times, and single-plex configuration have brought their use into question to complement the clinical information. Here, we report a chemiluminescence immunoassay (CLIA) for multiparametric quantification of specific IgE to penicillin G, penicillin V, amoxicillin, and piperacillin, using histone H1 as a carrier. The developed CLIA allowed the determination of BLC-specific IgE below 0.1 IU/mL, thus allowing identification of allergic patients with better sensitivity, using only 25 mu L of a sample (serum). The immunoassay was successfully applied in a cohort of 140 human serum samples, showing good sensitivity (64.6%) as well as specificity (100%), which significantly improve the predictive character of existing BLC-allergy in vitro tests.P.Q.C. acknowledges financial support from Generalitat Valenciana through the research staff-training program (GVA ACIF/2018/173). This research has been funded by the H2020 Program (project COBIOPHAD, grant agreement No. 688448), which is an initiative of the Photonics PPP (www.photonics21.org).Quintero-Campos, P.; Juárez-Rodríguez, MJ.; Morais, S.; Maquieira Catala, Á. (2020). Multiparametric Highly Sensitive Chemiluminescence Immunoassayfor Quantification of b-Lactam-Specific Immunoglobulin E. Analytical Chemistry. 92(21):14608-14615. https://doi.org/10.1021/acs.analchem.0c03020S14608146159221World Health Organization. WHO Report on Surveillance of Antibiotic Consumption; 2018, 27–28.Doña, I., Torres, M. J., Montañez, M. I., & Fernández, T. D. (2017). In Vitro Diagnostic Testing for Antibiotic Allergy. Allergy, Asthma & Immunology Research, 9(4), 288. doi:10.4168/aair.2017.9.4.288Ariza, A., Garzon, D., Abánades, D. R., de los Ríos, V., Vistoli, G., Torres, M. J., … Pérez-Sala, D. (2012). Protein haptenation by amoxicillin: High resolution mass spectrometry analysis and identification of target proteins in serum. Journal of Proteomics, 77, 504-520. doi:10.1016/j.jprot.2012.09.030Romano, A., & Warrington, R. (2014). Antibiotic Allergy. Immunology and Allergy Clinics of North America, 34(3), 489-506. doi:10.1016/j.iac.2014.03.003Ansotegui, I. J., Melioli, G., Canonica, G. W., Caraballo, L., Villa, E., Ebisawa, M., … Zuberbier, T. (2020). IgE allergy diagnostics and other relevant tests in allergy, a World Allergy Organization position paper. World Allergy Organization Journal, 13(2), 100080. doi:10.1016/j.waojou.2019.100080Han, M., Shin, S., Park, H., Park, K. U., Park, M. H., & Song, E. Y. (2013). Comparison of Three Multiple Allergen Simultaneous Tests: RIDA Allergy Screen, MAST Optigen, and Polycheck Allergy. BioMed Research International, 2013, 1-6. doi:10.1155/2013/340513Park, D. J., Lee, J., Kim, S.-Y., Kwon, H. J., Lee, H. K., & Kim, Y. (2019). Evaluation of AdvanSure AlloScreen Max Panel With 92 Different Allergens for Detecting Allergen-Specific IgE. American Journal of Clinical Pathology, 151(6), 628-637. doi:10.1093/ajcp/aqz023Bojcukova, J., Vlas, T., Forstenlechner, P., & Panzner, P. (2019). Comparison of two multiplex arrays in the diagnostics of allergy. Clinical and Translational Allergy, 9(1). doi:10.1186/s13601-019-0270-yWurpts, G., Aberer, W., Dickel, H., Brehler, R., Jakob, T., Kreft, B., … Brockow, K. (2020). Guideline on diagnostic procedures for suspected hypersensitivity to beta-lactam antibiotics. Allergologie select, 4(01), 11-43. doi:10.5414/alx02104eHan, X., Cao, M., Wu, M., Wang, Y.-J., Yu, C., Zhang, C., … Huang, W. (2019). A paper-based chemiluminescence immunoassay device for rapid and high-throughput detection of allergen-specific IgE. The Analyst, 144(8), 2584-2593. doi:10.1039/c8an02020ePark, K. H., Lee, J., Sim, D. W., & Lee, S. C. (2018). Comparison of Singleplex Specific IgE Detection Immunoassays: ImmunoCAP Phadia 250 and Immulite 2000 3gAllergy. Annals of Laboratory Medicine, 38(1), 23-31. doi:10.3343/alm.2018.38.1.23Kosnik, M., Silar, M., Zidarn, M., & Korosec, P. (2017). Comparison of the diagnostic value of two assays for specific immunoglobulin E in suspected beta lactam allergy. Medicinski pregled, 70(7-8), 209-215. doi:10.2298/mpns1708209kBlumenthal, K. G., Peter, J. G., Trubiano, J. A., & Phillips, E. J. (2019). Antibiotic allergy. The Lancet, 393(10167), 183-198. doi:10.1016/s0140-6736(18)32218-9Wong, T., Atkinson, A., t’ Jong, G., Rieder, M. J., Chan, E. S., & Abrams, E. M. (2020). Beta-lactam allergy in the paediatric population. Paediatrics & Child Health, 25(1), 62-62. doi:10.1093/pch/pxz179Stone, C. A., Trubiano, J., Coleman, D. T., Rukasin, C. R. F., & Phillips, E. J. (2019). The challenge of de‐labeling penicillin allergy. Allergy, 75(2), 273-288. doi:10.1111/all.13848Zhang, S., Zhang, Z., Shi, W., Eremin, S. A., & Shen, J. (2006). Development of a Chemiluminescent ELISA for Determining Chloramphenicol in Chicken Muscle. Journal of Agricultural and Food Chemistry, 54(16), 5718-5722. doi:10.1021/jf060275jLiu, Y.-C., Jiang, W., Chen, Y.-J., Xiao, Y., Shi, J.-L., Qiao, Y.-B., … Wang, Q. (2013). A novel chemiluminescent ELISA for detecting furaltadone metabolite, 3-amino-5-morpholinomethyl-2-oxazolidone (AMOZ) in fish, egg, honey and shrimp samples. Journal of Immunological Methods, 395(1-2), 29-36. doi:10.1016/j.jim.2013.06.006Zhao, Z., Batley, M., D’Ambrosio, C., & Baldo, B. A. (2000). In vitro reactivity of penicilloyl and penicillanyl albumin and polylysine conjugates with IgE-antibody. Journal of Immunological Methods, 242(1-2), 43-51. doi:10.1016/s0022-1759(00)00213-1Medical, S.; Diagnostics, S. IMMULITE 3gAllergy Specific IgE Universal Kit. 2015, 1–35.Phadiatop, I. ImmunoCAP Determining the Presence of IgE Anti- Bodies with ImmunoCAP Phadiatop in Appropriate Evaluation of Atopic Patients. 2013, 1–2.Garvey, L. H., Ebo, D. G., Mertes, P., Dewachter, P., Garcez, T., Kopac, P., … Scherer, K. (2019). An EAACI position paper on the investigation of perioperative immediate hypersensitivity reactions. Allergy, 74(10), 1872-1884. doi:10.1111/all.1382

The mutational landscape of myeloid leukaemia in down syndrome

Children with Down syndrome (DS) are particularly prone to haematopoietic disorders. Paediatric myeloid malignancies in DS occur at an unusually high frequency and generally follow a well-defined stepwise clinical evolution. First, the acquisition of mutations in the GATA1 transcription factor gives rise to a transient myeloproliferative disorder (TMD) in DS newborns. While this condition spontaneously resolves in most cases, some clones can acquire additional mutations, which trigger myeloid leukaemia of Down syndrome (ML-DS). These secondary mutations are predominantly found in chromatin and epigenetic regulators-such as cohesin, CTCF or EZH2-and in signalling mediators of the JAK/STAT and RAS pathways. Most of them are also found in non-DS myeloid malignancies, albeit at extremely different frequencies. Intriguingly, mutations in proteins involved in the three-dimensional organization of the genome are found in nearly 50% of cases. How the resulting mutant proteins cooperate with trisomy 21 and mutant GATA1 to promote ML-DS is not fully understood. In this review, we summarize and discuss current knowledge about the sequential acquisition of genomic alterations in ML-DS

Cinética química: Ley diferencial de velocidad

El objetivo principal de este artículo es el estudio de la velocidad de reacción, y los factores que la modifican. Este objetivo general se desglosa en los siguientes objetivos particulares: Conocer la velocidad de una reacción en función de la concentración de los reactivos Establecer como la velocidad de reacción se relaciona con la estequiometría de dicho proceso Introducir conceptos como, constante de velocidad y orden de reacción Determinar la Ley de velocidad experimentalmenteMorais Ezquerro, SB.; Atienza Boronat, MJ. (2019). Cinética química: Ley diferencial de velocidad. http://hdl.handle.net/10251/122934DE

Enzyme Inhibition Microassays on Blu-Ray Disks for Drug Discovery

[EN] An enzyme inhibition-based assay for drug discovery developed by microarraing on Blu-ray disks is presented. As a proof-of-concept, the system screens a selected molecule library of potential chemical inhibitors against the glycoenzyme peroxidase, identifying the promising lead compounds with high selectivity using standard Blu-ray disks and drives. In order to face the first drug discovery stages, we establish the bases for a high-throughput screening assay and a methodology based on hypersurfaces suitable to manage a high number of data as well.This work was supported by the Spanish Ministry of Economy and Competitiveness and the European Region Development Fund under award reference: CTQ2016-75749-R.Sancho-Fornes, G.; Peris Chanzá, EJ.; Giménez-Romero, D.; Morais, S.; Maquieira Catala, A. (2019). Enzyme Inhibition Microassays on Blu-Ray Disks for Drug Discovery. ACS Omega. 4(3):5595-5600. https://doi.org/10.1021/acsomega.8b03537S559556004

Termoquímica

Con este objeto de aprendizaje se explican conceptos básicos sobre los cambios de energía que acompañan a las reacciones químicas. Se definen los términos básicos de la Termoquímica, el concepto de calor y calor de reacción y trabajo. Se enuncia el primer principio de la termodinámica y se definen términos como el calor de reacción a volumen y a presión constante y entalpía de formación estándar.Morais Ezquerro, SB.; Atienza Boronat, MJ. (2018). Termoquímica. http://hdl.handle.net/10251/10435

Efecto de la temperatura sobre la velocidad de reacción: Ley de Arrhenius

Dado que la cinética es la parte de la química que mejor estudia la evolución de una reacción, conocer los factores que pueden modificar la velocidad es un objetivo primordial. El objetivo principal de este artículo es conocer cómo la temperatura afecta a la velocidad de reacción. Este objetivo general se desglosa en los siguientes objetivos particulares: ¿ Entender que la energía de activación es una barrera para que se produzca una reacción y que dicho impedimento se puede superar a medida que la temperatura aumenta ¿ Analizar por qué el número choques eficaces aumenta con la temperatura ¿ Calcular la constante de velocidad a otras temperaturas.Morais Ezquerro, SB.; Atienza Boronat, MJ. (2019). Efecto de la temperatura sobre la velocidad de reacción: Ley de Arrhenius. http://hdl.handle.net/10251/122836DE

Leyes integradas de velocidad

El objetivo principal de este artículo es el estudio de la velocidad de reacción. Este objetivo general se desglosa en los siguientes objetivos particulares: ¿ Determinar la velocidad a la que se produce una reacción química ¿ Establecer la relación entre constante de velocidad y constante de desaparición del reactivoMorais Ezquerro, SB.; Atienza Boronat, MJ. (2019). Leyes integradas de velocidad. http://hdl.handle.net/10251/122813DE

Espontaneidad de las reacciones químicas

En este objeto de aprendizaje se definen y explican conceptos como proceso reversible e irreversible, entropía y se enuncian la segunda y tercera ley de la termodinámica. El objeto está compuesto de ejemplos y actividades que el alumno debe de realizar para alcanzar los objetivos propuestos.Morais Ezquerro, SB.; Atienza Boronat, MJ. (2018). Espontaneidad de las reacciones químicas. http://hdl.handle.net/10251/104343DE