General health information, including leukocyte counts with differential; platelet counts; and hemoglobin, aspartate aminotransferase, alanine aminotransferase, blood urea nitrogen, creatinine, and C-reactive protein level measurements, was obtained for each participant from their laboratory studies. Total serum immunoglobulin E (IgE) levels were measured using the ImmunoCAP assay (Thermo Fisher Scientific, Uppsala, Sweden), and specific IgE levels were measured using a multiple antigen simultaneous test immunoblot assay for 61 inhalants (AdvanSure Alloscreen; LG Life Science, Seoul, Korea). Atopy was defined as a specific IgE level of ≥2 against >1 allergen or a total IgE level of 150 IU/mL.
Immunocap assay
Manufactured by Thermo Fisher Scientific
Sourced in Sweden, United States
The ImmunoCAP assay is a laboratory test designed to detect and measure specific antibodies in a person's blood. It is used to identify the presence and levels of immunoglobulin E (IgE) antibodies, which are associated with allergic responses. The ImmunoCAP assay provides quantitative results, allowing for the assessment of the patient's allergic sensitivity to various environmental and food-related allergens.
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Lab products found in correlation
31 protocols using immunocap assay
1
Immunological Assessment of Atopy
2
Quantifying Tissue Protein Levels
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As previously described,25 (link)26 (link) the protein concentrations of tissue extracts were determined using the Pierce 660nm Protein Assay Kit (ThermoFisher Scientific) and samples were thawed at room temperature and vortexed for thorough mixing. Tissue homogenates were then assayed for periostin proteins by using commercially available enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems, Minneapolis, MN, USA). Multiple cytokine analysis kits (IL-1ɑ, IL-1β, IL-5, IL-6, IL-10, IL-13, IL-17A, IL-22, IL-23, IL-33, chemokine [C-C motif] ligand [CCL]-11, CCL-24, chemokine [C-X-C motif] ligand [CXCL]-1, CXCL-2, CXCL-8, interferon (IFN)-γ, myeloperoxidase, transforming growth factor-β, S100A8, glycoprotein130, and B cell activating factor) were obtained from R&D Systems (Cat. No. LMSAHM), and data were collected using Luminex 100 (Luminex, Austin, TX, USA). Data analysis was performed using the MasterPlex QT version 2.0 (MiraiBio, Alameda, CA, USA). The levels of total IgE, Staphylococcal enterotoxins (SE)-specific IgE (SEA, SEB, and SEC), and eosinophil cationic protein (ECP) in nasal tissue homogenates were measured using the ImmunoCAP® assay (ThermoFisher Scientific). All assay procedures mentioned were run in duplicate according to the manufacturer's protocol. All the protein levels in the tissue homogenate were normalized to the concentration of total protein.
3
Longitudinal Antibody Response Monitoring
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Blood samples were collected at the following time‐points: at baseline (T0) and at different time‐points after the first injection: 6 hours (T0.25), 1 day (T1), 1 week (T7), 2 weeks (T14), 1 month (second dose, T28), 1 month and 2 hours (T28.08), 2 months (T56), 3 months (T84), 4 months (T112), 5 months (T140) and 8 months (follow‐up, T224). EDTA blood and gel‐separated serum were placed on ice immediately after venipuncture. Blood was also collected from nine self‐reported healthy controls for baseline analysis. All serum samples were allowed to coagulate for 60 minutes. Serum and plasma were obtained by centrifugation and stored at −80°C. Total IgE was determined using the ImmunoCap assay according the manufacturer's instructions (Thermo Fisher Scientific).
4
Quantifying Aspergillus Antibodies in Serum
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Serum samples were routinely tested for total specific A fumigatus immunoglobulin G using ImmunoCap assay (ImmunoCAPTM, Thermo Fisher, UK). Positive cut-off value of >40 mgA/L was used for all patients except for Cystic Fibrosis patients, whose cut-off was >90 mgA/L.
5
Inflammatory Marker Quantification in Tissue
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We selected 16 inflammatory markers based on previous reports.21 (link)23 (link) The protein concentrations of tissue extracts were determined using the Pierce 660 nm Protein Assay Kit (Thermo Fisher Scientific ., Waltham, MA, USA). Enzyme-linked immunosorbent assays for IL-22 and human neutrophil elastase (HNE) were performed with commercially available kits (all from R&D Systems, Minneapolis, MN, USA). Multiplex cytokine analysis kits (IL-1β, IL-5, IL-6, IL-8, IL-17A, IFN-γ, eotaxin-3, myeloperoxidase [MPO], matrix metalloproteinase [MMP]-9, TGF-β1, and periostin) were purchased from R&D Systems, and data were collected using a Luminex 100 reader (Luminex, Austin, TX, USA). Data analysis was conducted using MasterPlex QT version 2.0 (MiraiBio, Alameda, CA, USA). The levels of total IgE, specific IgE to staphylococcal enterotoxins (SE-IgE [SEA, SEB, SEC, and TSST-1]), and eosinophil cationic protein (ECP) in nasal tissue homogenates were measured using the ImmunoCAP assay (Thermo Fisher Scientific). A SE-IgE level > 0.35 IU/mL for at least one SE-specific IgE (SEA, SEB, SEC, and TSST-1) was defined as SE-specific IgE positive. All assay procedures mentioned were run in duplicate according to the manufacturer’s protocol. All protein levels in the tissue homogenates were normalized to the concentration of total protein. Further details are provided in the Supplementary Data S1 .
6
Milk Allergen-Specific Antibody Assay
7
Sera Analysis of Bovine Meat Allergy
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Sera were obtained from 31 patients with IgE antibodies to bovine meat at the Hospital Universitario La Paz (Madrid, Spain) after informed consent of the patients and with the approval by the Ethics Committee of La Paz University Hospital (Madrid, Spain) (EK565/2007). Patients with IgE antibodies to beef extracts in the ImmunoCap assay (Thermo Fisher Scientific, Uppsala, Sweden) were included in the study. The demographic, clinical, and serological characteristics of the patients were detailed in Table 1 . Interestingly, besides beef, some patients also had IgE antibodies to pork and/or lamb meat.
8
Allergy Serum Analysis of Korean Patients
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Serum samples were collected from 7 patients (average age, 30 years; age range, 12–59 years) who visited the Allergy-Asthma Center at Severance Hospital, Seoul, Korea (Table 1 ). Allergy diagnosis was based on patient history and a skin prick test. Specific IgE reactivity to Japanese hop was determined using an ImmunoCAP assay (Thermo Fisher Scientific, Uppsala, Sweden). Methods used to collect serum samples were approved by the Institutional Review Board of our institution (4-2017-1197).
9
Quantifying Serum Tryptase Levels
10
Quantifying Peanut-Specific IgE Antibodies
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Levels of specific IgE to peanut and peanut components Ara h 1, Ara h 2, Ara h 3, Ara h 6, Ara h 8, and Ara h 9 were quantified in patient plasma by using the ImmunoCAP assay (Thermo Fisher, Uppsala, Sweden) and the Phadia 100 analyzer according to the manufacturer’s instructions. Quantification of IgE to Ara h 1, Ara h 2, Ara h 3, Ara h 8, and Ara h 9 was performed at the time of recruitment and IgE to Ara h 6 at a later time, when the test became commercially available. Any values that indicated a specific IgE level greater than 100 kUA/L were diluted with specific IgE diluent (Thermo Fisher) 1 in 10 and retested.
The ImmunoCAP ISAC assays (Thermo Fisher) were performed according to the manufacturer’s instructions. The arrays were visualized by using a LuxScan 10K scanner (Core Life Sciences, Irvine, Calif) and analyzed by using Phadia Microarray Image Analysis software.
The ImmunoCAP ISAC assays (Thermo Fisher) were performed according to the manufacturer’s instructions. The arrays were visualized by using a LuxScan 10K scanner (Core Life Sciences, Irvine, Calif) and analyzed by using Phadia Microarray Image Analysis software.
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