Immunocap isac

Manufactured by Thermo Fisher Scientific

Sourced in Sweden, United States, Italy, Japan

ImmunoCAP ISAC is a laboratory diagnostic tool designed for the quantitative measurement of specific IgE antibodies in human serum or plasma samples. It utilizes an array-based technology to simultaneously detect and quantify a panel of allergen-specific IgE antibodies.

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Lab products found in correlation

Advia centaur platform, by Siemens (1 mentions) Prism v7, by GraphPad (1 mentions) Spss statistics v24, by IBM (1 mentions) Luxscan 10k, by CapitalBio (1 mentions) Phadia 100, by Thermo Fisher Scientific (1 mentions) Unicap 100, by Thermo Fisher Scientific (1 mentions) Luxscan 10k a, by CapitalBio (1 mentions) Isac technology, by Thermo Fisher Scientific (1 mentions) Immunocap feia, by Thermo Fisher Scientific (1 mentions) Genepix pro, by Molecular Devices (1 mentions) Genepix 4000b, by Molecular Devices (1 mentions)

Close spelling variants of this product

ImmunoCAP (382 citations) ImmunoCAP ISAC system (4 citations) ImmunoCAP ISAC 112 (3 citations) ImmunoCAP ISAC microarray (3 citations) ImmunoCAP ISAC technology (2 citations)

27 protocols using immunocap isac

Measurement of Allergen-specific IgE and IgG4

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Allergen‐specific IgE was measured using the ADVIA Centaur platform (Siemens Healthcare Diagnostics, Inc, Tarrytown, NY, United States) according to standard methods. Values provided are the sum of Phl p 1 and Phl p 5. Phl p 1‐ and p 5‐specific IgG4 was determined by ELISA.¹ All samples were tested by ImmunoCAP‐ISAC (Thermo Fisher Scientific, Uppsala, Sweden) to investigate sensitization profiles.
Statistical analysis was performed using IBM SPSS Statistics, v24. As data did not follow normality, Mann‐Whitney U test was applied to test the differences in the treatment at each time point. On the other hand, Friedman test was used to obtain differences due to the time in each treatment. Moreover, Wilcoxon test was performed for pair‐based comparisons. In all cases, P‐values < .05 were considered significant. Data were represented using violin plots showing sample median ± max and min values with Prism v7.0 software (GraphPad Software, La Jolla, CA).

Barker‐Tejeda T.C., Bazire R., Obeso D., Mera‐Berriatua L., Rosace D., Vazquez‐Cortes S., Ramos T., Rico M.D., Chivato T., Barbas C., Villaseñor A., Escribese M.M., Fernández‐Rivas M., Blanco C, & Barber D. (2020). Exploring novel systemic biomarker approaches in grass‐pollen sublingual immunotherapy using omics. Allergy, 76(4), 1199-1212.

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Characterization of Allergic Patient Samples

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Blood samples were obtained from grass and birch pollen allergic patients and non- allergic individuals after informed consent of the patients with the approval of the local ethics committee (Medical University of Vienna, Austria, EK 1641/2014). All experiments were performed in accordance with relevant guidelines and regulations. Grass and birch pollen allergy was diagnosed based on case history, skin testing and the detection of allergen-specific IgE antibodies by allergen microarray (ImmunoCAP ISAC, Thermofisher, Uppsala, Sweden). Table 1 provides a summary of the demographic and clinical characterization of the patients.
Rabbit IgG antibodies specific for Bet v 1 and Phl p 5 had been raised by immunization of rabbits with purified recombinant allergens (rBet v 1, rPhl p 5) or with KLH-coupled allergen-derived peptides using complete and incomplete Freund’s adjuvant (CFA, IFA), respectively (Charles River, Kislegg, Germany) as described^{43 (link),44}. Supplementary Tables 2 and 3 provide a summary of the specificities of the peptide-specific antisera^{43 (link),44}. The monoclonal mouse antibodies, Bip 1 and 4 A6 were raised against Bet v 1 and Bet v 2, respectively and are described^{28 (link),45 (link)}.

Najafi N., Hofer G., Gattinger P., Smiljkovic D., Blatt K., Selb R., Stoecklinger A., Keller W., Valent P., Niederberger V., Thalhamer J., Valenta R, & Flicker S. (2019). Fusion proteins consisting of Bet v 1 and Phl p 5 form IgE-reactive aggregates with reduced allergenic activity. Scientific Reports, 9, 4006.

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Serum IgE Profiling Using ImmunoCAP ISAC

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Capillary blood samples were obtained from the fingertip and incubated at room temperature for 15 min. After centrifugation at 14,000 rpm, serum was separated from the blood cells. Serum samples were subsequently stored at 4°C for transport and -20°C until further analysis. Analysis of sera for specific IgE to single purified allergens was done by means of the allergen multiplex array ImmunoCAP ISAC (Thermo Fisher Scientific, Uppsala, Sweden). According to the manufacturer’s protocol, the test was performed with 30 μl of serum (Protocol No. 20-01-02-6). The resulting fluorescent signals were measured with a confocal laser scanner (LuxScan-10K, CapitalBio, Beijing, China). Data were analyzed using Phadia Microarray Image Analyzer (MIA) software and transformed into semi-quantitative ISAC Standardized Units (ISU). Specific IgE values ≥0.3 ISU were considered positive. Participants with a positive value to any of the 112 allergens on the ImmunoCAP ISAC were considered as sensitized.

Stemeseder T., Schweidler B., Doppler P., Klinglmayr E., Moser S., Lueftenegger L., Himly M., Lang R., Zumbach J., Oostingh G.J., Hawranek T., Bathke A.C, & Gadermaier G. (2017). Exposure to Indoor Allergens in Different Residential Settings and Its Influence on IgE Sensitization in a Geographically Confined Austrian Cohort. PLoS ONE, 12(1), e0168686.

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Atopic Dermatitis Immune Profiling

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At the age of 36 months, blood is collected from all children still participating in the study to obtain serum, EDTA-blood and citrated blood (figure 1). A multiplex assay (ImmunoCAP ISAC, ThermoFisher, Waltham, USA) will be used to determine specific IgE against allergen components in serum, while differential blood counts for neutrophils, eosinophils, basophils, monocytes and lymphocytes are obtained from EDTA blood. In children diagnosed with AD and appropriate controls (sex-matched, age-matched, birthmonth, place of residence-matched), flow cytometric analysis and isolation of PBMCs are additionally performed from citrate blood analogous to cord blood (figure 2).

Preis S., Schmidt L., Tizek L., Schielein M., Lang V., Bleuel R., Duswald A., Sitaru S., Blasini A., Gasteiger C., Merdha L., Kurgyis Z., Kuschel B., Hauenstein E., Sander M., Niedermeier S., Argiriu D., Engel S., Skabytska Y., Silva R.L., Hils M., Evers B., Kaesler S., Hufnagel H., Köberle M., Amar Y., Zink A, & Biedermann T. (2022). Munich atopy prediction study (MAPS): protocol for a prospective birth cohort addressing clinical and molecular risk factors for atopic dermatitis in early childhood. BMJ Open, 12(9), e059256.

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Multiplex Measurement of Specific IgE Antibodies

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Total serum IgE levels were measured using Phadia 100; results were reported in kIU/L. Specific serum IgE antibodies were measured with the ImmunoCAP ISAC (Thermo Fisher Scientific, Waltham, MA USA). ImmunoCAP ISAC is a miniaturized immunoassay platform that allows for multiplex measurement of specific IgE antibodies to many allergen components using 20 μL of serum or plasma. Allergens are immobilized on a microarray chip to allow simultaneous measurement of specific IgE antibodies to 112 components from 51 allergen sources.^{21 (link)} Test results were measured with a biochip scanner and results were reported in ISAC standardized units (ISU) and categorized based on the manufacturer’s cutoff levels (< 0.3 ISU, undetectable or very low; 0.3-0.9 ISU, low; 1-14.9 ISU, moderate/high; and ≥ 15 ISU, very high). Values above 1 ISU were considered positive.

Siah K.T., Santosa A., Cheung C.K., Soh A.Y, & Bigliardi P.L. (2020). Atopic Patients Who Fulfilled Rome III Criteria for Irritable Bowel Syndrome Had Higher Animal Danders Sensitization. Journal of Neurogastroenterology and Motility, 26(2), 267-273.

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Pediatric Allergic Disorders and nsLTPs

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This study included 800 consecutive pediatric patients who visited the pediatric allergology clinic at the University of Campania “Luigi Vanvitelli” from 2010 to 2020. All patients were between the ages of 1 and 18 years old and were being followed for atopic disorders such as allergic asthma, atopic dermatitis, and allergic rhinitis. They had a suspected diagnosis of FA proposed by their primary care pediatricians. The study retrospectively analyzed the serum-specific IgE for eight different nsLTPs using the microarray method (ImmunoCAP ISAC, ThermoFisher Scientific, Milan, Italy). First, the data concerning nsLTPs were extracted and then compared. The nsLTPs analyzed were Ara h 9 (peanut), Jug r 3 (walnut), Cor a 8 (hazelnut), Pru p 3 (peach), Tri a 14 (wheat), Art v 3 (mugwort), Ole e 7 (olive tree), and Pla a 3 (plane tree). Sensitization was diagnosed in the presence of a value greater than 0.35 ISU-E.

Indolfi C., Dinardo G., Klain A., Contieri M., Umano G.R., Decimo F., Abbadessa S., Vitulano C., Ciprandi G, & Miraglia del Giudice M. (2023). Sensitization to nsLTP: A Retrospective Study in An Italian Pediatric Population over the Last Decade. Journal of Immunology Research, 2023, 4053799.

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Mite Allergen Specific IgE Characterization

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A pool of patients’ sera characterized using ImmunoCAP ISAC (Phadia, Thermo Fisher Scientific, Waltham, USA) a specific IgE levels of D. pteronyssinus and B. tropicalis were separately incubated overnight at 4 °C with different proteins concentration (0.1–10 µg/mL) of commercial extracts of Der p. and B. tropicalis. The allergenic proteins of D. pteronyssinus and B. tropicalis present on ImmunoCAP were Der p1, Der p2, Der p10, Der p 23 while Blo t5 was displayed on ISAC. The CAP-inhibition test was carried out with a specific programme in UniCap 100 (Phadia, Thermo Fisher Scientific, Waltham, USA) towards all 16 commercial allergenic extracts of D. pteronyssinus and B. tropicalis.

González-Pérez R., Poza-Guedes P., Barrios del Pino Y., Matheu V, & Sánchez-Machín I. (2019). Evaluation of major mite allergens from European standardized commercial extracts for in vivo diagnosis: addressing the need for precision medicine. Clinical and Translational Allergy, 9, 14.

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Longitudinal IgE Antibody Profiling

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We measured IgE to 112 components from 51 sources using ImmunoCAP ISAC (Thermo Fisher) at all 6 follow-ups. Levels of component-specific IgE antibodies were reported in ISAC standardized units. We discretized IgE data using a binary threshold (positive ≥0.30 ISAC standardized units).¹⁷ (link) We used the following annotations for component-specific IgE antibody responses: active components—we considered components to be active if ≥3 participants had a positive IgE response at each time point¹⁸ (link); and “drop-out” components—components that become inactive after having been active at an earlier time point.

Howard R., Belgrave D., Papastamoulis P., Simpson A., Rattray M, & Custovic A. (2018). Evolution of IgE responses to multiple allergen components throughout childhood. The Journal of Allergy and Clinical Immunology, 142(4), 1322-1330.

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Characterization of Phl p 1-specific IgE

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Recombinant allergens were purchased from Biomay (Vienna, Austria). The Phl p 1–specific IgE Fab clone 25 was isolated from a combinatorial library in a previous study (19 (link)) (Table I). The V regions of this clone were also grafted onto a complete human monoclonal IgG₁ Ab designated P1 IgG₁ and used to engineer ScFv 25 H chain variable (V_H)/25 L chain variable (V_L) (14 (link), 20 (link)). Serum samples from grass pollen–allergic patients were analyzed with approval of the Ethics Committee of the Medical University of Vienna in a retrospective and anonymized manner. The diagnosis of grass pollen allergy was based on a patient’s history of seasonal symptoms, skin prick testing, and allergen-specific IgE measurements as described (5 (link)). Grass pollen–allergic patients’ sera were also tested for IgE reactivity toward the four major recombinant grass pollen allergens, that is, Phl p 1, Phl p 2, Phl p 5, and Phl p 6, by ELISA. A comprehensive IgE reactivity profile toward >100 purified allergen molecules was established using ImmunoCAP ISAC assays (Thermo Fisher Scientific, Uppsala, Sweden) (21 (link)) (Supplemental Table I). Phl p 1–specific IgE Ab levels in patients’ sera were determined by ImmunoCAP g205 (Thermo Fisher Scientific). Phl p 1–derived peptides were synthesized and purified as described (22 (link)).

Madritsch C., Gadermaier E., Roder U.W., Lupinek C., Valenta R, & Flicker S. (2015). High-Density IgE Recognition of the Major Grass Pollen Allergen Phl p 1 Revealed with Single-Chain IgE Antibody Fragments Obtained by Combinatorial Cloning. Journal of immunology (Baltimore, Md. : 1950), 194(5), 2069-2078.

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Profiling Allergenic Sensitization Patterns

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We measured sIgE to 112 allergenic molecules using ImmunoCAP ISAC (Thermo Fisher Scientific-Phadia AB, Uppsala, Sweden) at the follow-up at age 11 years. The level of component-specific IgE antibodies was reported in ISAC Standardised Units (ISU). To ascertain co-occurring sensitisations among participants, we dichotomised IgE data according to the manufacturer's guidelines, using a binary threshold (positive>0.30 ISU). To evaluate the differential connectivity structure of component-specific IgEs, we used continuous raw values.

Fontanella S., Frainay C., Murray C.S., Simpson A, & Custovic A. (2018). Machine learning to identify pairwise interactions between specific IgE antibodies and their association with asthma: A cross-sectional analysis within a population-based birth cohort. PLoS Medicine, 15(11), e1002691.

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