Total serum IgE level was determined using the Immulite XPI, solidphase chemiluminescence assay (Siemens Medical Solutions, Malvern, PA, USA). IgE levels to pork and beef were measured using a fluoroenzyme immunoassay (Phadia Immuno CAP; courtesy of Platts-Mills laboratory Charlottesville, VA, USA).3 (link) To measure specific IgE, for galactose alpha-1,3-galactose (alpha-gal) we used an assay which employs a monoclonal antibody to cetuximab with a correlation of 0.95 to the commercially available assay in Europe which uses an antibody to beef thyroglobulin.16 (link) Values greater than 0.35 IU/mL were considered positive in both assays. sTryp levels were determined using a commercial fluoro-enzyme immunoassay (Phadia Immuno CAP, Uppsala, Sweden) at CLIA-approved laboratories, and levels above 11.4 ng/mL were considered elevated.
Immulite xpi
Manufactured by Siemens
Sourced in United States, Germany
The Immulite XPi is a laboratory equipment product developed by Siemens. It is an automated immunoassay system designed for the quantitative determination of various analytes in biological samples.
Automatically generated - may contain errors
Lab products found in correlation
End point technique, by Siemens (2 mentions)
Cobas 6000, by Roche (2 mentions)
Cobas 8000, by Roche (2 mentions)
Colorimetric enzymatic assay, by Merck Group (1 mentions)
Edta tube, by BD (1 mentions)
Ids isys instrument, by Immunodiagnostic Systems (1 mentions)
Advia 2120i, by Siemens (1 mentions)
Immulite 2000 xpi, by Siemens (1 mentions)
D 100, by Bio-Rad (1 mentions)
13 protocols using immulite xpi
1
Measuring Serum IgE and Alpha-gal Levels
2
Serum SHBG Measurement by ELISA
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Serum SHBG levels were measured using a human SHBG DuoSet solid-phase sandwich ELISA (R&D Systems, USA) according to the manufacturer’s instructions. The intra- and interassay coefficients of variation for serum SHBG were 2.8% and 5.1%, respectively. The DuoSet ELISA was validated against a chemiluminescent immunometric assay (Immulite XPi, Siemens, Germany) in eight samples. The intraclass correlation coefficient was 0.974 (95% CI 0.862, 0.995).
3
Biomarker Quantification in Blood
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Blood samples were collected in lithium-heparin, EDTA and Serum tubes. Lithium-heparin tubes (4.5 mL LH PSTTM II, Becton-Dickinson, NJ, America) were centrifuged at 1300 CRF for 10 min at room temperature (RT), plasma was frozen at −80 °C until it was analyzed for glucose concentrations. Glucose was measured by means of an end-point technique (Siemens, The Netherlands). EDTA tubes (8 mL, Becton-Dickinson, NJ, America) were centrifuged at 1200 G for 15 min at 4 °C, and plasma was frozen at −80 degrees until it was analysed for free fatty acids concentrations. Free fatty acids were assessed using an enzymatic test kit according to the manufacturer’s protocol (InstruChemie, Delfzijl, The Netherlands). Serum tubes (5 mL, Becton-Dickinson, NJ, USA) were set aside for at least 30 min, where after they were centrifuged at 1300 G for 10 min at RT, serum was frozen at −80 degrees until it was analysed for ketone content and cortisol concentration. Beta-hydroxybutyrate (βHB) was determined via a colorimetric enzymatic assay (Sigma-Aldrich; St. Louis, MO, USA). Analysis was performed according to manufacturer’s protocol. Cortisol was measured with immunometric chemiluminescence (sandwich) assay with Immulite XPi (Siemens, Den Haag, The Netherlands).
4
Serum Tryptase Levels in Clinical Assessments
5
Fasted Serum Biomarker Assessment
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Fasted venous blood samples were taken from the antecubital vein. The blood samples were allowed to clot for ∼30 min and centrifuged at 1000g for 15 min. Subsequently, aliquots of serum were stored at −80 °C, until further analysis. Serum samples were analyzed by the Central Diagnostic Laboratory at the Maastricht University Medical Centre (The Netherlands) for T3, as marker of energy availability, and vitamin D (25-(OH)D), procollagen 1 intact n-terminal propeptide (P1NP) and C-terminal telopeptide 1 (CTX—I) as markers of bone status. Total T3 was determined with a chemiluminescent immunoassay (Immulite XPi instrument, Siemens Healthcare Diagnostics). Intact P1NP, CTX—I, and 25(OH)D were measured using chemiluminescence immunometric assays on the IDS-iSYS instrument (Immunodiagnostic Systems Holdings, PLC, Tyne and Wear, UK). Reliability (CV) values for between-day measurements were ≤ 9 % (CTX—I), ≤5 % (P1NP), ≤10 % (T3), and ≤ 9 % (25(OH)D). Low T3 was defined as <0.8 nmol/L and low vitamin D was defined as <50 nmol/L. Low P1NP, or CTX-I were based on reference values according to age and gender (Supplemental Table 1 ).
6
Circulating Thyroid Hormones in Pups
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Blood samples were collected for analysis of circulating thyroid hormones [Thyroxine (T4), Triiodothyronine (T3), Thyroid stimulating hormone (TSH)], from all parental animals, pups on PND 4 and PND 13 (pooled sample per litter). Blood samples were put into tubes without anticoagulant for serum separation. The tubes were kept at room temperature and the serum was separated by centrifugation at 3000 rpm (4 °C) for 10 minutes and investigated using hormone analyzer (Immulite xpi, Siemens, Germany). The T4 for samples of adult males and pups on PND 13 were analyzed and significant differences were not observed in treatment group. Further assessment of T4 in blood samples from the dams and pups on PND 4 were not done. Also, other hormones were not measured (T3, TSH).
7
Serum biomarkers for allergy
8
Fasting Blood Biomarkers in VNS
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On the days of either scan, fasting blood (EDTA plasma and serum) was drawn right before the measurements were done. On the day of the VNS-off scan, blood samples were taken after VNS was switched off, right before injection of the tracer. Glucose was measured using a hexokinase-based assay (Cobas 6000, Roche Diagnostics, Mannheim, Germany) with a reference range of 3.1–7.8 mmol/L. Insulin was measured with Immulite XPi, Siemens Healthcare, Erlangen, Germany). Initial results were in mU/L and were converted to SI units with a factor of 6.00 (1 mU/L = 6.00 pmol/L) [18 ] with a reference range between 12 and 150 pmol/L. C-reactive protein (CRP) was measured using a turbidimetric assay (Cobas 8000) with a reference range of < 10 mg/L and a detection limit of > 1 mg/L.
9
Hematological Measurements and EPO Assay
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Erythrocyte, reticulocyte, white blood cell and platelet counts were determined with a Siemens Advia 2120i (Siemens Healthcare Diagnostics, Surrey, UK). Serum erythropoietin (EPO) was measured by a chemiluminescent assay on a Siemens Immulite XPi (Siemens Healthcare Diagnostics, Surrey, UK).
10
Plasma Biomarker Quantification Protocol
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All blood samples were immediately centrifuged at 1000 g at 4°C for 10 min, after which plasma was stored in −80°C until further analysis. Blood samples were analyzed for cortisol, creatine kinase, glucose, and insulin (Gelderse Vallei hospital, Ede, NL). Cortisol was measured with immunometric chemilumiescence (sandwich) assay with Immulite XPi, (Siemens, the Netherlands). Creatine kinase was measured using Vista device (Siemens, the Netherlands). Free fatty acids were assessed using an enzymatic test kit according to the manufacturer's protocol (InstruChemie, Delfzijl, Netherlands). Glucose was measured using absorption changes caused by the formation of NADH as a measure of glucose concentration and was measured bichromatic (340 and 383 nm) by means of an end point technique (Siemens, the Netherlands). Insulin was measured using an solid-phase enzyme-linked chemilumiescent immunometric assay with Immulite 2000 XPi (Siemens, the Netherlands).
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