Relevant bibliographies by topics / ELISA en sandwich

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  2. Dissertations / Theses
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Academic literature on the topic 'ELISA en sandwich'

Author: Grafiati
Published: 11 September 2021
Last updated: 14 February 2022

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Journal articles on the topic "ELISA en sandwich"

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Katsurada, Akemi, Yoshiaki Hagiwara, Kazuya Miyashita, Ryousuke Satou, Kayoko Miyata, Naro Ohashi, L. Gabriel Navar, and Hiroyuki Kobori. "Novel sandwich ELISA for human angiotensinogen." American Journal of Physiology-Renal Physiology 293, no. 3 (September 2007): F956—F960. http://dx.doi.org/10.1152/ajprenal.00090.2007.

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We recently reported that urinary excretion rates of angiotensinogen (UAGT) provide a specific index of intrarenal renin-angiotensin (ANG) system (RAS) status in ANG II-dependent hypertensive rats. When this is shown to be applicable to human subjects, a diagnostic test to identify those hypertensive patients most likely to respond to an RAS blockade could provide useful information to allow a mechanistic rationale for selection of an optimized approach to treatment of hypertensive patients. However, simple and accurate methods to measure human angiotensinogen (hAGT) are unavailable. For future studies of human subjects, we developed antibodies and a sensitive and specific quantification system for hAGT using a sandwich ELISA. We raised two antibodies against hAGT: a mouse monoclonal antibody and a rabbit polyclonal antibody. The standard curve of this ELISA exhibited a high linearity (0.31–20 ng/ml). The correlation coefficient was >0.99. Plasma angiotensinogen concentrations of healthy volunteers ranged from 28 to 71 μg/ml ( n = 10). The ratio of UAGT to urinary creatinine concentration ranged from 5.0 to 30 μg/g ( n = 7). Intra- and interassay coefficients of variation ranged from 4.4 to 5.5% and from 4.3 to 7.0%, respectively. This ELISA system had no cross-reactivity with major proteins in proteinuric urine samples, such as human albumin, immunoglobulin, or transferrin. Moreover, the cross-reactivity of the system with angiotensin peptides was also negligible. This hAGT ELISA will be a useful tool to investigate the relationship of UAGT and reactivity to antihypertensive drugs in hypertensive patients.
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Zhao, Weilin, Seinyone Pao, Fatima Malik, James Soh, Sonalis Fernandez, and William J. Chirico. "A Sandwich ELISA for Phosphoglycerate Kinase." Journal of Immunoassay and Immunochemistry 29, no. 3 (June 2, 2008): 220–33. http://dx.doi.org/10.1080/15321810802119588.

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Ibrahim R B Aly, Samir A M Zaahkouk, Alaa A M Samn, Ibrahim A E Zahran, and Sawsan A M EL-Shamy. "Effective diagnosis of schistosomiasis haematobium by Silver beads Sandwich ELISA." International Journal of Research in Pharmaceutical Sciences 11, SPL4 (December 21, 2020): 1398–404. http://dx.doi.org/10.26452/ijrps.v11ispl4.4312.

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Schistosomiasis is a major public health problem. Diagnosis by simple and rapid immunoassays is a priority. The magnetic bead immunoassay using magnetic nanoparticles conjugated with anti-schistosomal antibody was evaluated for di­agnosing human schistosomiasis infection. The present study was to evaluate the sandwich ELISA as a simple test for the detection of schistosomal antigen (CSA) in serum and urine samples of S. haematobium patients and compare it with ELISA. Investigation conducted on eighty six cases divided to three groups, 34 were positively for Schistosoma haematobium, 32 were positively for intestinal parasites ova and negative for S. haematobium ova in urine and 20 were negative urine and stool examination (Control). Immunomagnetical bead based Enzyme-linked immunosorbent assay (ELISA) using for detected for antigen in sera and urine infected by S. haematobium. Sandwich ELISA sensitivities was 79.4% (serum) and 73.5% (urine) and which increase by used nano-sandwich ELISA to 88.2% (serum) and 82.4% (urine), respectively. Sandwich ELISA specificities was 86.4% (serum) and 80.8% (urine) and which increase by used nano-sandwich ELISA to 93.3% (serum) and 88.5% (urine). We found that, nano-sandwich ELISA assay had highly sensitive and specifically and technical method was applicably, fast, cheaper, accurate and promising diagnostic method for schistosomiasis.
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Rhee, Jong Il, Jörg Hagedorn, Thomas Scheper, and Karl Schügerl. "Flow-injection sandwich ELISA for bioprocess monitoring." Journal of Automated Methods and Management in Chemistry 21, no. 4 (1999): 121–25. http://dx.doi.org/10.1155/s1463924699000140.

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A fully automated flow-injection immunoassay based on sandwich enzyme-linked immunosorbent assay (ELISA) is described for the model system: protein G-sepharose, rabbit IgG and horseradish peroxidase (HRP)-labelled protein A. After injecting rabbit IgG and HRP-labelled protein A into a cartridge containing protein G-sepharose sequentially, a mixture of hydrogen peroxide and the redox indicator, 2.2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS) is passed through the cartridge. The HRP-labelled protein A bound in the cartridge is directly proportional to the concentration of rabbit IgG. The colour variation of ABTS caused during the reaction between HRP and H202in the cartridge is detected photometrically. The whole assay procedure is controlled and evaluated by a computer. Rabbit IgG and HRP-labelled protein A are also detected by a fluorometer, which is introduced into the flow system. In the flow-injection sandwich ELISA, the slope of the calibration curve is 0.4491 in the range of 0 and 300 μg ml-1rabbit IgG, while it is 0.1274 in the heterogeneous immunoassay. So the flow-injection sandwich ELISA system is found to be more sensitive than a heterogeneous immunoassay for the monitoring of the model protein.
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Huang, Ruo-Pan, Ying Lin, Li-Pai Chen, Weimin Yang, and Ruochun Huang. "ELISA-based Protein Arrays: Multiplexed Sandwich Immunoassays." Current Proteomics 1, no. 3 (July 1, 2004): 199–210. http://dx.doi.org/10.2174/1570164043379226.

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Oka, T., M. Ito, T. Egashira, N. E. Miller, and H. Hattori. "Plasma PLTP concentration measured by sandwich ELISA." Atherosclerosis 151, no. 1 (July 2000): 220. http://dx.doi.org/10.1016/s0021-9150(00)81001-1.

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Rhee, Jong Il, Jorg Hagedorn, Thomas Scheper, and Karl Schugerl. "Flow-injection sandwich ELISA for bioprocess monitoring." Journal of Automated Methods & Management in Chemistry 21, no. 4 (July 1, 1999): 121–25. http://dx.doi.org/10.1080/146392499300613.

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Lim, Yoon, Jin-Hua Zhong, and Xin-Fu Zhou. "Development of mature BDNF-specific sandwich ELISA." Journal of Neurochemistry 134, no. 1 (April 21, 2015): 75–85. http://dx.doi.org/10.1111/jnc.13108.

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Panchal, Manoj, K. Muralidhar, and S. K. Gupta. "A Sensitive Sandwich ELISA for Buffalo Prolactin." Journal of Immunoassay and Immunochemistry 28, no. 2 (March 22, 2007): 147–54. http://dx.doi.org/10.1080/15321810701212062.

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VANREE, R., S. STAPEL, and R. AALBERSE. "Reverse-sandwich ELISA may underestimate antibody titers." Journal of Allergy and Clinical Immunology 84, no. 4 (October 1989): 562–63. http://dx.doi.org/10.1016/0091-6749(89)90371-0.

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Dissertations / Theses on the topic "ELISA en sandwich"

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Nerson, Daniel Alain Jacques. "Méthode ELISA "sandwich" pour la détection rapide d'antigènes solubles bactériens." Lyon 1, 1985. http://www.theses.fr/1985LYO1W032.

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Reyes, Valdez Devi. "DETERMINACIÓN DE NIVELES SÉRICOS DE I-CAM Y V-CAM EN MUESTRAS DE PACIENTES Y VALIDACIÓN DEL PROCESO MEDIANTE ELISA SÁNDWICH." Tesis de Licenciatura, Universidad Autónoma del Estado de México, 2019. http://hdl.handle.net/20.500.11799/104993.

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Antecedentes: ICAM-1 y VCAM-1 pertenecen a un grupo de proteínas de tipo inmunoglobulina y se expresan sobre las células del endotelio vascular. La expresión de ICAMs es inducible e incrementa durante la activación celular, así el endotelio inflamado se caracteriza por un incremento en la expresión de ICAM-1 y de VCAM.1. El ensayo inmunoabsorbente ligado a enzimas (ELISA) utiliza una enzima como marcador para medir la formación de complejos antígeno-anticuerpo. El marcador enzimático que se emplea en estos análisis se conjuga con un ligando, que puede ser un antígeno, un anticuerpo específico para el antígeno de interés o un anticuerpo para el anticuerpo primario. La validación es la confirmación, mediante evidencia objetiva, de que se cumplen los requisitos de un método para la utilización o aplicación específica prevista. Objetivo: Determinar los niveles de moléculas de adhesión molecular I-CAM-1 y V-CAM-1 en pacientes; y realizar la validación del proceso mediante la técnica de ELISA Sándwich. Materiales y Métodos: Elaborar una curva de calibración para ICAM-1 y VCAM-1, medir muestras de pacientes aleatorios y determinar la muestra más alta, la cual será utilizada para realizar la validación, se llevan a cabo las pruebas de linealidad, repetibilidad y reproducibilidad. Resultados: Para la prueba de linealidad se obtuvieron los siguientes resultados: 1) VCAM-1 se obtuvo una r2= 0.99 dentro del intervalo 193 ng /mL a 1550.808 ng /mL, con un % de recobro entre 93-100 % 2) ICAM-1 se obtuvo una r2= 0.99, dentro del intervalo 160.173 ng /mL a 1256.808 ng /mL con un % de recobro entre 93-102 %. Para la prueba de repetibilidad 1) VCAM-1 se obtuvo un coeficiente de variación de 0.66 % - 2.73 %. 2) ICAM-1 se obtuvo un coeficiente de variación de 2.29 % - 4.86 %. En la prueba de reproducibilidad para 1) VCAM-1 se obtuvo un coeficiente de variación de 2.27 %, 2) ICAM-1 se obtuvo un coeficiente de variación de 3.90 %. Todas las pruebas son satisfactorias en ambos casos. Conclusiones: Dentro de los intervalos obtenidos en las pruebas de linealidad el método se comporta de manera lineal. De acuerdo con los resultados de repetibilidad y reproducibilidad podemos afirmar que el método es preciso. Dado el porcentaje de recuperación o recobro obtenido, podemos afirmar que el método es exacto. Los resultados obtenidos respecto a linealidad, precisión y exactitud, la técnica de Elisa Sándwich es adecuada para determinación de niveles séricos de ICAM-1 y VCAM-1.
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Woldu, Haddish Haben. "Analys av C3a och sC5b-9 med sandwich-ELISA för att mäta komplementaktivering vid subklinisk borrelios." Thesis, Linnéuniversitetet, Institutionen för kemi och biomedicin (KOB), 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-76120.

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Lyme borreliosis (LB) is caused by spirocheter of Borrelia burgdorferi sensu lato. There are different types of borrelia species and some differ in their ability to survive in the presence of the complement system. B. afzelii is complementresistant while B. garinii is complementsensitive. This is based on the ability to recruit immune regulators, such as factor H to the bacterial surface and prevent activation of the complement system. Some individuals may show anti-Borrelia antibodies without having developed clinical symptoms. This may indicate a more effective immune response against spirochetes. The aim of this study was to investigate differences in complement activation by measuring C3a and sC5b-9 with sandwich ELISA between two previously Borrelia-exposed groups; individuals with previous subclinical Lyme borreliosis (SB) and patients previously diagnosed with neuroborreliosis (NB), and a control group without signs of LB exposure. Samples analyzed in this study consisted of controls (Ctrl, n = 8st), SB (n = 60st) and NB (n = 22st). Plasma from the groups were activated with ACA1 and Lu59. To compare the relative increase between the groups, complement factor C3a and the soluble terminal complement complex, sC5b-9, were analyzed using sandwich-ELISA.The analysis of C3a and sC5b-9 showed higher activation with Lu59 than ACA1, which is consistent with previous studies. According to C3a-analysis, no significant differences were observed between the groups for neither ACA1 nor Lu59. According to sC5b-9-analysis, a significant difference between SB and Ctrl (p= 0,0081) for Lu59 was observed. Conclusion of the studie was that further studies are required to interpret how this complement activation affects LB from a clinical prespective.
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Echevarria, Del Valle Rinaldo Josué. "Desarrollo de un kit ELISA tipo sandwich, para la detección de antígenos de excreción-secreción de Fasciola hepatica linnaeus, 1758." Bachelor's thesis, Universidad Nacional Mayor de San Marcos, 2013. https://hdl.handle.net/20.500.12672/10591.

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La fasciolosis, producida por Fasciola hepatica, es una parasitosis cosmopolita de importancia médico-veterinaria que afecta a la ganadería lechera con prevalencias superiores al 80% que generan significativas pérdidas económicas. Por lo tanto, es importante contar con técnicas de diagnóstico que permitan la identificación temprana del parásito como las técnicas de captura cuando el antígeno se encuentra en baja concentración en la muestra. El objetivo del presente trabajo fue desarrollar un kit de ELISA para detectar antígenos de excreción-secreción de Fasciola hepatica. Se desarrolló un kit de ELISA tipo sandwich para la detección de antígenos de excreción-secreción de Fasciola hepatica, el cual alcanzó un límite de detección de 100 ng/mL, empleando una concentración óptima de anticuerpo de captura (anticuerpos policlonales de ratón) y segundo anticuerpo (anticuerpos policlonales de conejo) de 10 y 5 µg/mL, respectivamente; y una dilución óptima del conjugado (anticuerpo monoclonal anti-inmunoglobulinas de conejo conjugado con la enzima peroxidasa) de 1/1000. El kit de ELISA tipo sandwich, presento un valor de sensibilidad del 100%, una especificidad del 96.6% y valores predictivos positivos y negativos de 50% y 96.6% respectivamente; con relación al examen coproparasitológico directo. Al comparar el ELISA tipo sandwich con la contrainmunoelectroforesis (CIEF) se obtuvo una especificidad de 93.5% y un valor predictivo negativo del 100%. El kit de ELISA tipo sandwich desarrollado permite detectar antígenos metabólicos en muestras de heces de ganado ovino.
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VEYRES, NOU PATRICIA. "Detection des igg liees aux plaquettes et libres circulantes par un dosage elisa de type sandwich : application aux thrombopenies des sujets v.i.h. seropositifs." Nice, 1988. http://www.theses.fr/1988NICE6802.

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Karim, Ali Hussein. "Mätning av kluven Kaspas-3 och kluven PARP i manganbehandlade prostatacancerceller." Thesis, Högskolan Kristianstad, Fakulteten för naturvetenskap, 2019. http://urn.kb.se/resolve?urn=urn:nbn:se:hkr:diva-19079.

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Prostatacancer är den sjätte mest förekommande cancertypen i världen, och den tredje vanligaste cancertypen bland män. De olika typer av behandlingar som finns idag botar oftast inte sjukdomen. Det är därför viktigt att utveckla bättre behandlingsmetoder. Det är sedan tidigare känt att mangan kan orsaka apoptos i olika celltyper. Det ger en möjlighet att använda mangan för att hämma cancer och det är därför viktigt att veta vilka apoptotiska markörer som är involverade. Syftet med projektet var att undersöka om de sker en ökning av de apoptotiska markörerna kluven Kaspas-3 och kluven PARP efter manganbehandling av prostatacancerceller. Därefter kunna avgöra om manganbehandlingen har orsakat celldöd genom att inducera apoptos. Under projektet odlades prostatacancerceller (PC3) som sedan behandlades med 200 µM mangan under 6, 24 och 48 timmar. Därefter mättes mängden av proteinerna kluven Kaspas-3 och kluven PARP med hjälp av ett sandwich-ELISA kit. En tydlig stegvis ökning av apoptosmarkörerna med inkuberingstid hade förväntats. Det förväntade resultatet erhölls inte, för kluven Kaspase-3 ledde manganbehandlingen t.o.m. till en sänkning av koncentrationen. Det kan ha uppstått problem vid analysen som t.ex dålig lysering av cellerna eller ojämn tillväxt av dem. Det behövs fler studier för att utreda detta och för att undersöka andar potentiella apoptosmarkörer.
Prostate cancer is the sixth most common cancer type in the world, and the third most common cancer type among men. The different types of treatments that are available today do not usually cure the disease. It is therefore important to develop better treatment methods. It has previously been stated that manganese can cause apoptosis in different cell types. It provides an opportunity to use manganese to inhibit cancer and it is therefore important to know which apoptotic markers that are involved. The purpose of the project was to investigate whether an increase in the apoptotic markers cleaved Kaspas-3 and cleaved PARP after manganese treatment of prostate cancer cells. Thereafter, it can determine whether manganese treatment has caused cell death by inducing apoptosis. During the project prostate cancer cells (PC3-cells) were cultivated, then treated with 200 µM manganese for 6, 24 and 48 hours. The amount of the proteins cleaved Kaspas-3 and cleaved PARP was measured by using a sandwich ELISA kit. A clear gradual increase of apoptotic markers with incubation time was expected. The expected result was not obtained, for cleaved Kaspase-3 manganese treatment even decreased the concentrations. There may have been problems with the performance of the analysis such as poor lysis of the cells or uneven growth of the cells. More studies are required to investigate this and other potential apoptotic markers.
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Classon, Lisa. "Atypiskt terminalt komplementkomplex : Kvantifiering av in vivo-nivåer av atypiskt terminalt komplementkomplex under normala och patofysiologiska betingelser." Thesis, Linnéuniversitetet, Institutionen för kemi och biomedicin (KOB), 2018. http://urn.kb.se/resolve?urn=urn:nbn:se:lnu:diva-78509.

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Slutsteget i immunförsvarets komplementaktivering innefattar en klyvning av komplementprotein C5, till C5a och C5b, vilket initierar bildandet av terminalt komplementkomplex (TCC) som i form av membran-attack-komplex (MAC) bildar cytotoxiska porer i bland annat gramnegativa bakterier. Bildandet av MAC kan blockeras av endogena regulatorer och TCC frisätts då som ett lösligt komplex, sC5b-9, i plasma. I examensarbetet studerades en variant av TCC, som i tidigare studier visats bildas oberoende av C3- och C5-konvertas när serum surgjorts till pH < 7,0 in vitro. Syftet med studien var att studera om detta atypiska TCC (aTCC) bildades hos grisar, som i en mekonium aspirationssyndrom (MAS)-modell, erhållit ett sänkt systemiskt pH in vivo. I syftet ingick också att etablera en ELISA-baserad metod för att analysera aTCC. I en sandwich ELISA användes monoklonal anti-C5a/C5a (desArg) (klon T13/9) som fångande antikropp och monoklonal anti-C9 (klon aE11) som detekterande antikropp för att analysera aTCC i plasmaprover från 18 MAS-grisar, samt i ett kontrollmaterial bestående av grisserum som surgjorts till pH 6,8 och 6,4 in vitro. Mängden aTCC i kontrollproverna ökade när pH sänktes men innehållet av aTCC i plasmaproverna minskade över MAS-studiens förlopp. När den relativa förändringen i aTCC relaterades till MAS-grisarnas slutliga pH kunde ett signifikant samband ses (p = 0,02) som visade att en större förändring i aTCC sammanföll med ett lägre slutligt pH. Nivåerna av aTCC var generellt sett högre i plasmaproverna jämfört med kontrollproverna vilket skulle kunnat bero på skillnader i plasma vs serum avseende aTCC eller att proverna kom från grisar med olika ålder och vikt. Avsaknad av grisspecifik standard och negativ kontroll samt lågt signal/brusförhållande bidrar till felkällor för analysen och denna kräver fortsatt optimering.
The late steps of complement activation involves a cleavage of complement protein C5, to C5a and C5b, which initiates the formation of terminal complement complex (TCC). The final complex is referred to as the membrane-attack-complex (MAC) which forms cytotoxic pores in, inter alia, gram-negative bacteria. The formation of MAC can be inhibited by endogenous regulators and the TCC is then released as a soluble complex, sC5b-9, in plasma. In the degree project, another type of TCC was studied, which in previous studies had shown to form independently of C3 and C5 convertases when serum was acidified to pH <7.0 in vitro. The purpose of the study was to investigate whether this atypical TCC (aTCC) was formed in piglets, which in a model of meconium aspiration syndrome (MAS), received a reduced systemic pH in vivo. The purpose was also to establish an ELISA for analyzing aTCC. Sandwich ELISA, with monoclonal anti-C5a / C5a (desArg) (clone T13/9) as a capture antibody and monoclonal anti-C9 (clone aE11) as a detection antibody, was used to analyse aTCC in plasma samples from 18 MAS piglets, and in control samples consisting of pig serum acidified to pH 6.8 and 6.4 in vitro. The amount of aTCC in the control samples increased when the pH was lowered, but the content of aTCC in the plasma samples decreased over the course of the MAS study. When the relative change in aTCC was related to the final pH of the MAS pigs, a significant relationship could be seen (p = 0.02) which showed that a major change in the aTCC coincided with a lower final pH. aTCC were generally higher in plasma samples compared with control samples, which could be due to differences in plasma vs serum for aTCC or that the samples came from pigs of different age and weight. Lack of pig-specific standard and negative control as well as low signal to noise ratio contribute to sources of error for the analysis and this requires continued optimization.
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Spindeldreier, Doerthe C. [Verfasser]. "Expression von caninem IL-4 in E.coli und Säugerzellen (CHO-Zellen/canine Knorpelzellen) sowie Entwicklung eines direkten Sandwich-ELISA zur Messung von caninem Interleukin-4 / Doerthe C. Spindeldreier." Berlin : Freie Universität Berlin, 2007. http://d-nb.info/1021939064/34.

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León, janampa Nancy. "Development of a test associated with magnetic nanoparticles for the diagnosis of tuberculosis." Thesis, Bordeaux, 2019. http://www.theses.fr/2019BORD0272.

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Mycobacterium tuberculosis provoque l'une des maladies qui présentent le taux de mortalité et de morbidité le plus élevé des Amériques et du monde. Dans les pays en développement, le diagnostic de la tuberculose (TB) repose sur la microscopie des frottis et des cultures bactériologiques. La première méthode a une faible sensibilité et la seconde met plusieurs semaines à atteindre un diagnostic de confirmation. L'absence de diagnostic rapide compromet les efforts de lutte contre la tuberculose, favorisant ainsi sa transmission à la population vulnérable. Actuellement, les nanoparticules magnétiques (MNP) fonctionnalisées par des biomolécules sont utilisées en biomédecine, en raison de leurs propriétés magnétiques, électriques et optiques. De cette manière, en appliquant des champs magnétiques externes, les MNP bio-fonctionnalisées sont utilisées pour détecter et concentrer les cellules et les biomolécules à partir d'échantillons biologiques.Dans ce travail, nous présentons la synthèse, la caractérisation et la bio-fonctionnalisation de nanoparticules magnétiques afin de développer un test ELISA en sandwich associé aux MNP, afin de détecter les antigènes de M. tuberculosis. À cette fin, des nanoparticules magnétiques ont été synthétisées par une méthode de co-précipitation. La surface des MNP a été amino-silanisée (MNP@Si@NH2) et caractérisée par des méthodes physico-chimiques.Les antigènes MTB évalués dans cette étude étaient: Hsp16.3, CFP10, ESAT6, MTC28, MPT64, protéine de 38 kDa, Ag85B et MoeX. Le clonage et l'expression de protéines recombinantes ont été réalisés dans le système de E. coli BL21 (DE3) pLysS. Des anticorps polyclonaux ont été produits chez des lapins Nouvelle-Zélande et des souris BALB/C, préalablement immunisés avec des antigènes recombinants purifiés. Des anticorps spécifiques (ab) ont été immobilisés sur les surfaces des MNP amino-silanisées. Les MNP@Si@ab ont été associées dans un test ELISA sandwich colorimétrique pour capturer et détecter les antigènes natifs du MTB à partir d’échantillons d’expectorations.La XRD, la spectroscopie Mössbauer, le potentiel zêta, la TEM et le FTIR ont validé l'obtention des MNP montrant un diamètre de cristal de diffraction de ~ 12,5 nm (10,48 ± 2,56 nm), une charge nette superficielle de +23,57 ± 2,87 mV, des profils caractéristiques de magnétite et une structure sphérique. De plus, une saturation en aimantation de 37,06 emu.g-1 a été observée. Pour la fonctionnalisation des surfaces de nanoparticules avec des anticorps, une méthode via l'utilisation d'ester activé (agent de couplage EDC/NHS) a été utilisée pour la formation de liaisons peptidiques. Des paramètres tels que le temps d'incubation, la concentration des agents de couplage et le niveau de saturation de surface des MNP amines silanisées (MNP@Si@NH2) ont déjà été standardisés.Enfin, des anticorps fonctionnalisés sur des MNP ont été utilisés pour capturer et détecter les antigènes recombinants et natifs de M. tuberculosis dans un test sandwich ELISA-MNP@Si@ab (dans un temps de réaction <4 h). Les antigènes ESAT6 et CFP10 ont été mieux différenciés dans les expectorations des patients atteints de tuberculose (fold value ~ 1,8). L'utilisation de MNP@Si@ab a amélioré la détection des antigènes du MTB dans des échantillons biologiques. Nos résultats sont encourageants, mais des évaluations supplémentaires sont nécessaire telles que la détermination de réactions croisées avec des échantillons d'expectorations provenant de patients atteints d'autres infections, la réalisation du test avec les expectorations fraîches de patients tuberculeux et la détermination de la sensibilité et de la spécificité de la méthode
Mycobacterium tuberculosis causes one of the diseases with the highest mortality and morbidity rate in the Americas and around the world. In developing countries, the diagnosis of tuberculosis (TB) is based on smear microscopy and bacteriological cultures. The first method has low sensitivity, and the second take several weeks to reach a confirmatory diagnosis. The lack of a rapid diagnosis compromises the efforts to control TB, favoring its transmission to the susceptible population. Currently, magnetic nanoparticles (MNPs) functionalized with biomolecules have been used in biomedicine, due the magnetic, electrical and optical properties. In this way, applying external magnetic fields, bio-functionalized MNPs is used to detect and concentrate cells and biomolecules from biological samples.In this work we present the synthesis, characterization and bio-functionalization of magnetic nanoparticles, to develop a sandwich ELISA assay associated to MNPs to detect antigens from M. tuberculosis. For this purpose, magnetic nanoparticles were synthesized by co-precipitation method. The MNP surface was amine-silanized (MNP@Si@NH2) and characterized by physical-chemical methods.The MTB antigens evaluated in this study were: Hsp16.3, CFP10, ESAT6, MTC28, MPT64, 38 kDa protein, Ag85B and MoeX. Cloning ad expression of recombinant proteins were made in E. coli BL21 (DE3) pLysS system. Polyclonal antibodies were produced in New Zealand rabbits and BALB/C mice, previously immunized with purified recombinant antigens. Specific antibodies (ab) were immobilized in the amine-silanized MNP surfaces. The MNP@Si@ab were associated in a colorimetric sandwich ELISA assay to capture and detect native MTB antigens from sputum samples.The XRD, Mössbauer spectroscopy, zeta potential, TEM and FTIR demonstrated the successful preparation of the MNPs showing a diffraction crystal diameter of ~12.5 nm (10.48 ± 2.56 nm), superficial net charge of ᶎ: +23.57 ± 2.87 mV, characteristic patterns of magnetite and a spherical structure. Additionally, a magnetization saturation of 37.06 emu.g-1 was observed. For the functionalization of nanoparticle surfaces with antibodies, active ester method (coupling agent EDC/NHS) were used for peptide bond formation. Parameters such as time of incubation, concentration of coupling agents and surface saturation level of amine-silanized MNPs (MNP@Si@NH2), were previously standardized.Finally, antibody functionalized on MNPs were used to capture and detect recombinant and native M. tuberculosis antigens in an ELISA-MNP@Si@ab sandwich test (in a reaction time <4 h). The ESAT6 and CFP10 antigens were better discriminated in sputum pooles from patients with TB (fold value ~ 1.8). The use of MNP@Si@ab improved the detection of MTB antigens in biological samples. Our results are encouraging, but the essay requires additional evaluations such as determining cross-reactions with sputum samples from patients with other infections, performing the test with fresh sputum of TB patients, and determining the sensitivity and specificity of the method
Mycobacterium tuberculosis causa una de las enfermedades con la tasa más alta de mortalidad y morbilidad en las Américas y en todo el mundo. En países en vías de desarrollo, el diagnóstico de tuberculosis (TB) se basa en microscopía de frotis y cultivos bacteriológicos. El primer método tiene baja sensibilidad y el segundo toma varias semanas para llegar a un diagnóstico confirmatorio. La falta de un diagnóstico rápido compromete los esfuerzos para controlar la TB, lo que favorece su transmisión a la población susceptible. Actualmente, las nanopartículas magnéticas (MNP) funcionalizadas con biomoléculas se han utilizado en biomedicina, debido a las propiedades magnéticas, eléctricas y ópticas. De esta manera, aplicando campos magnéticos externos, se utilizan MNP bio-funcionalizadas para detectar y concentrar células y biomoléculas a partir de muestras biológicas. En este trabajo presentamos la síntesis, caracterización y bio-funcionalización de las nanopartículas magnéticas para desarrollar un ensayo ELISA sándwich usando MNPs para detectar antígenos de M. tuberculosis. Para este propósito, las nanopartículas magnéticas fueron sintetizadas por el método de co-precipitación. La superficie de MNP fue amino-silanizada (MNP@Si@NH2) y se caracterizada por métodos físico y químicos. Los antígenos de MTB evaluados en este estudio fueron: Hsp16.3, CFP10, ESAT6, MTC28, MPT64, proteína de 38 kDa, Ag85B y MoeX. La clonación y la expresión de las proteínas recombinantes se realizaron en el sistema de E. coli BL21 (DE3) pLysS. Se produjeron anticuerpos policlonales en conejos de Nueva Zelanda y ratones BALB/C, inmunizados previamente con antígenos recombinantes purificados. Se inmovilizaron anticuerpos específicos (ab) en las superficies de MNP amino-silanizadas (MNP@Si@ab). El MNP@Si@ab fue utilizado en un ensayo ELISA sándwich colorimétrico para capturar y detectar antígenos de MTB nativos en muestras de esputo. La XRD, espectroscopia de Mössbauer, la potencial zeta, TEM y FTIR demostraron la preparación exitosa de los MNP, el cual mostró un diámetro de difracción del cristal de ~ 12.5 nm (10.48 ± 2.56 nm), carga neta superficial de ᶎ: +23.57 ± 2.87 mV, patrones característicos de magnetita y una estructura esférica. Además, una saturación de magnetización de 37.06 emu.g-1 fue observada. Para la funcionalización de superficies de nanopartículas con anticuerpos, se utilizó el método del éster activo para la formación de enlaces peptídicos. Parámetros tales como el tiempo de incubación, la concentración de los agentes de acoplamiento y el nivel de saturación de la superficie de las MNPs aminosilanizadas (MNP@Si@NH2) fueron estandarizadas. Finalmente, se usaron MNP funcionalizados con anticuerpos para capturar y detectar antígenos nativos y recombinantes de M. tuberculosis en una prueba sándwich de ELISA-MNP@Si@ab en un tiempo de reacción <4 h. Los antígenos ESAT6 y CFP10 se discriminaron mejor en las muestras de esputo de los pacientes con TB (fold value ~ 1,8). El uso de MNP@Si@ab mejoró la detección de antígenos de MTB en muestras biológicas con respecto a un sELISA convencional. Nuestros resultados son alentadores, pero el ensayo requiere evaluaciones adicionales, como determinar reacciones cruzadas con muestras de esputo de pacientes con otras infecciones, realizar la prueba con esputo frescos de pacientes con TB y determinar la sensibilidad y especificidad clînica del método
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Marassa, Ana Maria. "Identificação de fonte sanguínea em dípteros da Família Culicidae, em áreas de epizootia da febre amarela silvestre." Universidade de São Paulo, 2009. http://www.teses.usp.br/teses/disponiveis/6/6132/tde-20072009-153444/.

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A importância em conhecer o padrão alimentar em mosquitos da Família Culicidae permite esclarecer alguns aspectos relacionados à transmissão de zoonoses e estimar o grau de contato humano-vetor que é fator relevante em estudos epidemiológicos. Com o objetivo de explorar o comportamento alimentar dessa Família, em área epizoótica de febre amarela silvestre, foram coletados exemplares nos municípios de Santo Antônio das Missões e Garruchos, Estado do Rio Grande do Sul. Fêmeas ingurgitadas foram obtidas por aspiração em ambiente de mata, no período de setembro de 2005 a abril de 2007 e identificadas segundo fonte de sangue ingerido através da técnica imunoenzimática ELISA de captura no sistema avidinabiotina. Foram testadas seis fontes de alimento: ave, bovino, eqüino, humano, macaco e rato. Os resultados obtidos mediante a padronização de anticorpos monoclonais possibilitaram demonstrar pela primeira vez o reconhecimento de sangue humano ingerido nesses mosquitos pelo emprego da subclasse IgG1 e comprovar a sensibilidade e especificidade da técnica ELISA de captura. No município de Santo Antônio das Missões, de um total de 190 amostras, 60,9% reagiram para sangue de boi, 23,6% para humano, 9,9% para ave, 1,9% para macaco e 3,7% para combinações de dois hospedeiros. Quanto às amostras referentes ao município de Garruchos, das 158 fêmeas capturadas na área Cachoeirinha pode-se observar reatividade para ave (16%), boi (29,6%), humano (36,8%), cavalo (4%), macaco (0,8%) e combinações de hospedeiros (12,8%), enquanto que para as 149 fêmeas pertencentes à área de São José, detectou-se sangue ingerido de boi em (51,5%), ave e humano (11,5%), macaco (6,2%), cavalo (0,8%) e mistos (18,5%). Aedes scapularis, Aedes crinifer, Culex (Culex) spp., Haemagogus leucocelaenus apresentaram maior número de fêmeas ingurgitadas nos dois municípios. Os resultados obtidos com Aedes scapularis sugerem ecletismo, conforme combinações detectadas em amostras de sangue de diferentes fontes. Haemagogus leucocelaenus apresentou a maior proporção de amostras contendo sangue humano em relação às demais fontes e essa característica traz implicações, por ser espécie incriminada na transmissão e por se tratar de área de ocorrência de epizootias de febre amarela.
The knowledge of mosquitoes Culicidae host feeding patterns permits to clarify some aspects related to zoonosis transmission and to estimate the degree of human-vector contact which is relevant in epidemiological studies. Aiming to explore the feeding behavior of these mosquitoes, specimens were collected in the municipalities of Santo Antônio das Missões and Garruchos, Rio Grande do Sul, an epizootic area of sylvatic yellow fever. Engorged females were collected by aspiration from forested areas from September 2005-April 2007 and their blood meals were identified using the avidin-biotin system of immunoenzymatic ELISA capture. Six blood meal sources were tested: bird, cattle, horse, human, monkey and rat. The result achieved with the species-specific IgG1 mAb was unprecedented for mosquito blood meal identification and reinforced the sensibility and specificity of the immunoenzymatic ELISA capture. Of the 190 samples from Santo Antônio das Missões, 60.9% reacted to cattle, 23.6% to human, 9.9% to bird, 1.9% to monkey and 3.7% to mixed blood meals. In Garruchos, of the 158 females collected in Cachoeirinha, 16.0% reacted to bird, 29.6% to cattle, 36.8% to human, 4.0% to horse, 0.8% to monkey and 12.8% to mixed blood, while of the 149 engorged females from São José, blood from cattle accounted for 51.5%, of blood identified, bird and human 11.5%, monkey 6.2%, horse 0.8% and mixed blood 18.5%. Blood engorged females of Aedes scapularis, Aedes crinifer, Culex (Culex) spp., Haemagogus leucocelaenus predominated in the two municipalities. The results obtained with Aedes scapularis suggests its eclecticism, according to the combinations of blood which were detected from different sources. Haemagogus leucocelaenus was found to have the highest proportion of samples containing human blood in comparison with other sources, which has implications, on account of being incriminated in the transmission and also for taking into consideration the outbreaks reported that underline the risk of yellow fever.
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Books on the topic "ELISA en sandwich"

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Castro-Perez, Jose Miguel. "Sandwich" ELISA for prostatic-specific antigen (PSA) complexed to alpha-1 -antichymotrypsin. 1995.

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Book chapters on the topic "ELISA en sandwich"

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Ireland, H. Elyse, and John H. H. Williams. "Measuring Hsp72 (HSPA1A) by Indirect Sandwich ELISA." In Methods in Molecular Biology, 145–53. Totowa, NJ: Humana Press, 2011. http://dx.doi.org/10.1007/978-1-61779-295-3_12.

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Stanker, Larry H., and Robert M. Hnasko. "A Double-Sandwich ELISA for Identification of Monoclonal Antibodies Suitable for Sandwich Immunoassays." In Methods in Molecular Biology, 69–78. New York, NY: Springer New York, 2015. http://dx.doi.org/10.1007/978-1-4939-2742-5_7.

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Tang, Qiushi, Alisha M. Gruntman, and Terence R. Flotte. "Quantification of Total Human Alpha-1 Antitrypsin by Sandwich ELISA." In Methods in Molecular Biology, 211–16. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-7163-3_20.

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Borel, Florie, Qiushi Tang, and Christian Mueller. "Quantification of Z-AAT by a Z-Specific “Sandwich” ELISA." In Methods in Molecular Biology, 223–26. New York, NY: Springer New York, 2017. http://dx.doi.org/10.1007/978-1-4939-7163-3_22.

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Gonzalez, Rachel M., Susan M. Varnum, and Richard C. Zangar. "Sandwich ELISA Microarrays: Generating Reliable and Reproducible Assays for High-Throughput Screens." In Biomarker Methods in Drug Discovery and Development, 273–90. Totowa, NJ: Humana Press, 2008. http://dx.doi.org/10.1007/978-1-59745-463-6_13.

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Sylvester, I., A. J. Suffredini, R. L. Danner, and E. J. Leonard. "Effect of Diluents and Capture Antibodies on Detection of NAP-1 by Sandwich Elisa." In Advances in Experimental Medicine and Biology, 206. Boston, MA: Springer US, 1993. http://dx.doi.org/10.1007/978-1-4615-2952-1_51.

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Kimura, A., T. Uda, S. Nakashima, M. Osawa, H. Ikeda, S. Yasuda, and T. Tsuji. "ABO Blood Grouping and Species Identification of Bloodstains by Sandwich ELISA Using Monoclonal Antibody Specific for Human Erythrocyte Band 3." In Advances in Forensic Haemogenetics, 437–39. Berlin, Heidelberg: Springer Berlin Heidelberg, 1992. http://dx.doi.org/10.1007/978-3-642-77324-2_132.

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Ball, Hywel J., and David Finlay. "Diagnostic Application of Monoclonal Antibody (MAb)-Based Sandwich ELISAs." In Mycoplasma Protocols, 127–32. Totowa, NJ: Humana Press, 1998. http://dx.doi.org/10.1385/0-89603-525-5:127.

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Servoss, Shannon L., Rachel Gonzalez, Susan Varnum, and Richard C. Zangar. "High-Throughput Analysis of Serum Antigens Using Sandwich ELISAs on Microarrays." In Methods in Molecular Biology, 143–50. Totowa, NJ: Humana Press, 2009. http://dx.doi.org/10.1007/978-1-60327-811-9_10.

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"Sandwich ELISA." In Encyclopedia of Cancer, 3334. Berlin, Heidelberg: Springer Berlin Heidelberg, 2011. http://dx.doi.org/10.1007/978-3-642-16483-5_5155.

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Conference papers on the topic "ELISA en sandwich"

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Fang, Juan, Chaoyin Chen, and Shenglan Zhao. "Quantitative sandwich ELISA for the determination of walnut protein in foods." In International conference on Human Health and Medical Engineering. Southampton, UK: WIT Press, 2014. http://dx.doi.org/10.2495/hhme131042.

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Karachaliou, Chrysoula-Evangelia, Persefoni Klimetzou, Maria Paravatou-Petsotas, Ourania Tsitsilonis, Wolfgang Voelter, Hubert Kalbacher, Christos Zikos, and Evangelia Livaniou. "Development of an IgG/IgY sandwich-ELISA for the bioactive polypeptide Prothymosin alpha." In 35th European Peptide Symposium. Prompt Scientific Publishing, 2018. http://dx.doi.org/10.17952/35eps.2018.230.

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Bloom, J. W., J. A. Berkner, and G. Mitra. "FACTOR VIII:C, 80 KD AND 90-210 KD POLYPEPTIDE QUANTITATION BY SLISA." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644042.

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Purified Factor VIII:c is generally found proteolyzed into nultiple polypeptides. An 80 kD polypeptide appears to form a metal-linked complex with each of a series of 90-210 kD polypeptides and separation of these complexes results in loss af Factor VIII:c activity. To quantitate total Factor VIII:c content - active and inactive - three sandwich ELISA's were developed using an immunopurified polyclonal antibody as the capture antibody. This antibody was shown by a Western blot of Factor VIII:c to interact with all the 80-210 kD polypeptides. Utilizing biotinylated polyclonal antibody, an ELISA was developed that detected both the purified carboxy terminal (80 kD) and amino terminal (90-210 kD) polypeptides. Thrombin treated 80 kD polypeptide and Factor VIII:c were also detected by this ELISA. A second ELISA was developed with a biotinylated monoclonal, designated C8, that detected purified 90-210 kD polypeptides, but not the 80 kD polypeptide. A third ELISA was developed with a biotinylated monoclonal, designated C7F7, that detected only the purified 80 kD functional subunit of Factor VIII:c. Thrombin treated 80 kD polypeptide and Factor VIII:c were not detected by this ELISA. These three ELISA methods combined allow estimates of amounts of denatured or thrombin degraded Factor VIII:c to be determined, information not obtainable by the coagulation activity assays.
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Bitzer, A., A. Laber, E. Gadermaier, J. Wallwitz, G. Berg, and G. Himmler. "Sandwich ELISA for the Quantification of Soluble Human Semaphorin 4D, a factor promoting skeletal metastases." In OSTEOLOGIE 2019. Georg Thieme Verlag KG, 2019. http://dx.doi.org/10.1055/s-0039-1679968.

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IANNONE, G., P. G. DAVOLI, A. TRALDI, C. D'URSO, S. ARRIGHI, and R. P. REVOLTELLA. "IMMUNOLOGICAL CHARACTERIZATION OF ANTI-FACTOR VIII ANTIBODIES BY A SANDWICH- ELISA AND THE BIA-TECHNOLOGY." In Proceedings of the 7th Italian Conference. WORLD SCIENTIFIC, 2002. http://dx.doi.org/10.1142/9789812776457_0045.

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Yoo, Eunjeong, Yvonne Lin, Nicos Petasis, Augustin Garcia, Stan Louie, Isaac Asante, Eugene Zhou, and Song Ah Chae. "Abstract 534: Development and validation of sandwich ELISA to quantify circulating GRP78 as a cancer biomarker." In Proceedings: AACR 106th Annual Meeting 2015; April 18-22, 2015; Philadelphia, PA. American Association for Cancer Research, 2015. http://dx.doi.org/10.1158/1538-7445.am2015-534.

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Sugihara, T., J. Takamatsu, T. Kamiya, H. Saito, K. Kimata, and K. Kato. "A SENSITIVE ENZYME-LINKED IMMUNOSORBENT ASSAY(ELISA) FOR SERUM LAMININ." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643554.

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Laminin, a large glycoprotein, is a major and specific component of basement membrane. There were little or no circulating laminin in normal persons, although some recent reports have showed increased values in various diseases(ex. diabetes mellitus and liver disease) by a radioimmunoassay(RIA) using laminin fragment. The minimum detectable sensitivity of RIA was reported to be 20 ng/ml of serum sample. We describe here a more sensitive immunoassay system, and also the concentrations of laminin in sera from healthy subjects and patients. A sandwich ELISA method for measurement of laminin was established by use of purified antibodies to mouse laminin. The assay system consisted of poly-stylene balls with immobilized antibody F(ab’)2 fragments and the same antibody Fab1 fragments labeled with β-D-galactosidase from E.Coli. The assay was highly sensitive and can detect as small as 0.5 ng/ml of serum laminin.Coefficients of variation in within-run and between-run precision studies for serum laminin were good. Serum laminin levels in healthy subjects of various ages ranged from 1.5 to 3.9 ng/ml(n=60). Fifty eight patient sera (collagen disease(n=18), hepatic disease(n=20), and renal disease (n=20)) were examined. Significant differences between the normal sera and 3 diseases sera were observed as shown belowIt is concluded that circulating laminin apparently exists in normal persons and there are higher laminin level in some diseases which appears to involve basement membrane-rich organ
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Multani, Gitanjali, Priyanka Multani, Preston B. Landon, and Ratneshwar Lal. "Abstract DPOC-010: EARLY DETECTION: QUANTIFICATION OF B7-H4 VIA MODIFIED SANDWICH ELISA USING TRANSMISSION ELECTRON MICROSCOPY (TEM)." In Abstracts: 11th Biennial Ovarian Cancer Research Symposium; September 12-13, 2016; Seattle, WA. American Association for Cancer Research, 2017. http://dx.doi.org/10.1158/1557-3265.ovcasymp16-dpoc-010.

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Lecander, I., and B. Åstedt. "OCCURRENCE OF PAI-2 IN MEN AND NON-PREGNANT WOMEN." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1644458.

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PAI-2 is a specific plasminogen activator inhibitor most likely produced in the trophoblasts of placenta but presumably also in certain macrophages. It occurs in two molecular froms of 48 and 60 kDa. From placenta homogenates mainly the LMW-form can be isolated. During pregnancy mainly the HMW-form increases in the blood and disappears after delivery.We examined blood samples from 120 male blood donors (females excluded to avoid unknown pregnancies) and 20 staff members, 4 males and 16 females for the presence of PAI-2.PAI-2 antigen levels were measured with a sandwich ELISA using a polyclonal and a monoclonal antibody. The concentration was given in per cent of that in pooled term plasma.PAI-2 antigen was detected in 5 of the blood donors 9, 12, 22, 31 and 60 %. In one male staff member the antigen concentration was 9 % and in one female 72 %. Repeated analyzes of this last member for six months resulted in values between 30-90 %. Immunoblotting using a monoclonal antibody against PAI-2 showed a HMW-band of about 80 kDa.
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Pelzer, H., R. Egbring, R. Seitz, H. B. Blanks, M. Cazzola, and N. Heimburger. "THROMBIN-ANTITHROMBIN III COMPLEX - A NEW PARAMETER FOR DETECTION OF THROMBOTIC STATES." In XIth International Congress on Thrombosis and Haemostasis. Schattauer GmbH, 1987. http://dx.doi.org/10.1055/s-0038-1643674.

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Recently, we have developed an enzyme-linked immunosorbent assay (ELISA) for determination of thrombin-antithrombin III complex (TAT) in numan plasma (1). The test system follows the sandwich principle and uses two different antibodies directed against human thrombin and human antithrombin III, respectively. The antibodies bind selectively to the corresponding antigen moieties of TAT. The assay was calibrated with definite concentrations (2.0 to 60 μg/l) of preformed purified TAT added to TAT-poor plasma. Plots of absorbance 492 nm against TAT concentration revealed a linear correlation (r = 0.98). The lower limit of sensitivity of the assay was 0.5 μg/l. Mean coefficients of variation of 4 % (intraassay) and 7.5 % (interassay) were found for TAT concentrations between 2 and 60 μg/l. A reference range from 0.85 to 3.0 μg/l was calculated from TAT concentration in plasma samples from 88 healthy donors (mean value ± SD: 1.45 ± 0.4 μg/l). In plasma samples from patients with pulmonary embolism confirmed by lung-scan (n = 17), TAT concentrations between 10 and 20 μg/l were measured. In patients with deep vein thrombosis confirmed by phlebography (n = 15), TAT were found up to 7 - 13 μg/l. Patients with septicaemia associated with a consumption coagulopathy (n = 10), and patients with acute hepatic failure (n = 5) showed markedly increased TAT values. In plasma samples from patients with various metastatic malignant diseases, TAT concentrations of 30 to 60 μg/l could be measured. From these data we conclude that measurement of TAT can be a sensitive parameter for specific detection of a latent activation of the clotting pathway.
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