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Immunocap 100

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The ImmunoCAP 100 is an automated instrument used for in vitro diagnostic testing. It is designed to perform quantitative and qualitative analysis of specific IgE antibodies in human serum or plasma samples.

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26 protocols using immunocap 100

1

Peanut-Specific IgE and IgG4 Quantification

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Peanut- and peanut component-specific IgE and IgG4 were measured via ImmunoCAP 100 (Thermo Fisher, Uppsala, Sweden) according to the manufacturer’s specifications.
Burk C.M., Kulis M., Leung N., Kim E.H., Burks A.W, & Vickery B.P. (2016). Utility of component analyses in subjects undergoing sublingual immunotherapy for peanut allergy. Clinical and experimental allergy : journal of the British Society for Allergy and Clinical Immunology, 46(2), 347-353.
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2

Serum and Plasma Biomarker Extraction

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Collection: Fresh blood (2–3 mL) and serum were collected in EDTA-coated anticoagulant tube and pro-coagulant tube respectively, before being transported to the laboratory at low temperature.
Processing: (I) the pro-coagulant tubes were centrifuged at 1,000 g at 4 °C for 5 min. The top 500 µL of serum was sent for measurement of total immunoglobulin E (tIgE) and dust mite specific IgE (sIgE; ImmunoCAP-100®, Thermo Fisher Scientific, Sweden), and the remainder was stored at –80 °C. (II) The anticoagulant tubes were centrifuged at 800 g at 4 °C for 5 min, and the plasma was stored at –80 °C. The remaining cells were centrifuged in a Ficoll-Paque density gradient, and the layer of peripheral blood mononuclear cells (PBMC) was collected and stored at –80 °C.
Zhou J., Li W., Wen X., Zeng D., Lin J., Chen S., Zang N., Deng Y., Xie X., Ren L, & Liu E. (2021). Establishing a patient registry study database of dust mite allergic asthma in children: design, methodology and preliminary exploration. Annals of Translational Medicine, 9(12), 993.
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3

Peanut Allergy Immunology Protocol

Cited 3 times
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Blood for mechanistic studies was collected at baseline, annually, and at desensitization and SU food challenges. Serum pn-sIgE and pn-sIgG4 levels were assessed by using the ImmunoCAP 100 (Thermo Scientific, Waltham, Mass), as previously described.17 (link),18 (link) Ratios of pn-sIgG4 to pn-sIgE were calculated by first converting both IgG4 and IgE levels into micrograms per liter and then dividing pn-sIgG4 quantities by pn-sIgE quantities. A conversion factor of 2.42 µg/L = 1 kU/L was used for pn-sIgE. Basophil activation was assessed by using whole blood in the presence of IL-3 with several dilutions of crude peanut extract (103, 102, 101, and 100 ng/mL), anti-IgE (103 ng/mL), and media alone. Basophils were identified as CD123+CD203c+Lin− (CD3, CD14, CD19, and CD41) events, with CD63 as the primary marker of activation. Results were analyzed as a ratio of peanut-specific to nonspecific (anti-IgE) activation, as previously described.19 (link)
Kim E.H., Yang L., Ye P., Guo R., Li Q., Kulis M.D, & Burks A.W. (2019). Long-term sublingual immunotherapy for peanut allergy in children: Clinical and immunologic evidence of desensitization. The Journal of allergy and clinical immunology, 144(5), 1320-1326.e1.
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4

Peanut-Specific IgE and IgG4 Measurement

Cited 1 time
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Total IgE was measured by immunoassay, and peanut-specific-IgE (PN-IgE) and peanut-specific IgG4 (PN-IgG4) were measured using the ImmunoCAP 100 (Thermo Fisher Scientific, Waltham, MA) at baseline and at weeks 29, 44, 68 and annually at the time of OFC, as previously reported.11 (link)
Burks A.W., Wood R.A., Jones S.M., Sicherer S.H., Fleischer D.M., Scurlock A.M., Vickery B.P., Liu A.H., Henning A.K., Lindblad R., Dawson P., Plaut M, & Sampson H.A. (2015). Sublingual immunotherapy for peanut allergy: Long-term follow-up of a randomized multicenter trial. The Journal of allergy and clinical immunology, 135(5), 1240-8.e1-3.
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5

Comprehensive Blood Evaluation Protocol

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Blood tests for most patients also included complete blood count with differential analysis, C-reactive protein (CRP), basal serum tryptase (ImmunoCAP 100 Thermo Fisher Scientific, Uppsala, Sweden), and total IgE (Table 2).
Bizjak M., Košnik M., Terhorst-Molawi D., Dinevski D, & Maurer M. (2021). Cold Agglutinins and Cryoglobulins Associate With Clinical and Laboratory Parameters of Cold Urticaria. Frontiers in Immunology, 12, 665491.
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6

Quantifying Allergen-Specific IgE Levels

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Total IgE (kU/L), allergen serum sIgE (kUA/L) and component sIgE were measured with the ImmunoCAP assay (ImmunoCAP-100®, Thermo Fisher Scientific, Sweden), following the manufacturer’s instructions. The sIgE tests included Der p, Der f, Blomia tropicalis (Blo t), cat dander, dog dander, egg white, milk, cockroach, shrimp, and crab, along with the Der p allergen components, Der p1 and Der p2. A sIgE level over 0.35 kUA/L was defined as positive. Positive IgE tests were categorized into 6 classes: class 1 (≥0.35 to <0.70 kUA/L), class 2 (≥0.70 to <3.50 kUA/L), class 3 (≥3.50 to <17.50 kUA/L), class 4 (≥17.50 to <50.00 kUA/L), class 5 (≥50.00 to <100.00 kUA/L), and class 6 (≥100.00 kUA/L).
Zeng D., Li W., Zhou J., Wen X., Chen S., Xie X., Zang N., Deng Y., Ren L., Rizvi S.A., Shimizu Y., Park C.S., Khaltaev N, & Liu E. (2020). Analysis of the immunoglobin E molecular sensitization profile in children with allergic asthma and predictive factors for the efficacy of allergy immunotherapy. Annals of Translational Medicine, 8(21), 1459.
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7

Basophil Activation and Enumeration

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Detailed methodology is described in the Methods section in this article's Online Repository. Briefly, expression of CD63 and CD203c (markers of basophil activation) and enumeration of basophils (CD123+HLA-DR− cells), lymphocytes, and polymorphonuclear leukocytes (PMNs) were determined by means of flow cytometry, as previously described.20 (link), 21 (link), 22 (link) In samples from patients with peanut allergy, we determined the absolute basophil count using a similar methodology, with basophils identified as chemoattractant receptor–homologous molecule expressed on TH2 lymphocytes (CRTH2)–positive CD303−CD123+ cells.23 (link)
FCER1A, CPA3, and HDC gene expression was analyzed in whole-blood samples (PAXgene; PreAnalytiX, Hombrechtikon, Switzerland), as previously described.22 (link)
We measured serum concentrations of CCL2, CCL5, CCL11, IL-3, and TSLP by using ELISA, according to the manufacturers' instructions (Quantikine; R&D Systems, Minneapolis, Minn and Abcam, Cambridge, United Kingdom). For IL-3 measurements, we also performed spiking experiments (see the Methods section in this article's Online Repository). We measured serum total tryptase (α+β) levels with the ImmunoCAP 100 (Thermo Fisher, Uppsala, Sweden); tryptase concentrations that exceeded 11.4 μg/L were considered increased.
Korosec P., Turner P.J., Silar M., Kopac P., Kosnik M., Gibbs B.F., Shamji M.H., Custovic A, & Rijavec M. (2017). Basophils, high-affinity IgE receptors, and CCL2 in human anaphylaxis. The Journal of Allergy and Clinical Immunology, 140(3), 750-758.e15.
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8

Peanut SLIT Immunoassay Protocol

Cited 2 times
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Blood for immune studies was collected at baseline; after 6, 12, 24, 36, 48 months of peanut SLIT; and after SLIT avoidance. Serum Pn-sIgE and peanut-specific IgG4 (Pn-sIgG4) levels were measured using the ImmunoCAP 100 (Thermo Scientific, Waltham, MA), as previously described.15 (link), 23 (link) Ratios of Pn-sIgG4 to Pn-sIgE were calculated using a conversion factor of 2.42 mcg/L = 1 kU/L for Pn-sIgE.
, & Meade T.W. (2001). Design and intermediate results of the Lower Extremity Arterial Disease Event Reduction (LEADER)* trial of bezafibrate in men with lower extremity arterial disease [ISRCTN41194621]. Current Controlled Trials in Cardiovascular Medicine, 2(4), 195-204.
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9

Skin Prick and IgE Testing for Allergen Sensitivity

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Sensitization was assessed at baseline by skin prick testing, and for those unable to be skin tested because of wheezing at the time of the visit or because they could not discontinue antihistamines, IgE testing was performed to the major indoor allergens (cockroach, mouse urine, dust mite, cat, and dog) using the ImmunoCAP100 (ThermoFisher, Uppsala, Sweden). Skin prick testing was performed with a multitester device to 14 allergens (Multitest II, Lincoln Diagnostics, Decatur, Illinois). A positive skin test was defined as a net orthogonal wheal size ≥ 2mm.
Grant T., Lilley T., McCormack M.C., Rathouz P.J., Peng R., Keet C.A., Rule A., Davis M., Balcer-Whaley S., Newman M, & Matsui E.C. (2022). Indoor environmental exposures and obstructive lung disease phenotypes among asthmatic children living in poor, urban neighborhoods. The Journal of allergy and clinical immunology, 151(3), 716-722.e8.
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10

IgE Antibody Measurement for D. pteronyssinus

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Specific IgE antibodies to D. pteronyssinus (d1) (cut-off > 0.35 kU/L) were measured according to the manufacturer’s instructions on an ImmunoCAP 100 instrument (Thermo Fisher Scientific, Uppsala, Sweden).
Pauli G., Wurmser C., Roos A., Kokou C., Huang H.J., D’souza N., Lupinek C., Zakzuk J., Regino R., Acevedo N., Caraballo L., Vrtala S, & Valenta R. (2023). Frequent IgE recognition of Blomia tropicalis allergen molecules in asthmatic children and young adults in equatorial Africa. Frontiers in Immunology, 14, 1133935.
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