Quantification of Plasma Hormones

Plasma insulin was measured using an in-house radioimmunoassay, using an in-house-generated guinea pig anti-human insulin antibody and ¹²⁵I-labelled human insulin tracer. ¹²⁵I-labelled human insulin tracer was generated using ¹²⁵I (PerkinElmer Nederland) and human insulin (Novo Biolabs cat. no. 471). In this assay, bound–free separation is performed by second antibody/polyethylene glycol precipitation of antibody-bound insulin. The assay is calibrated on World Health Organization international standard 83/500. The cross-reactivity in this method is approximately 60% for insulin aspart and 50% for insulin lispro. The cross-reactivity for insulin degludec is not well known. Catecholamines were analysed by an LC-MS/MS method developed and validated in-house after derivatisation with propionic anhydride and subsequent solid-phase extraction [26 (link)]. Plasma cortisol was determined using a routine analysis method with an electrochemiluminescent immunoassay on a Cobas E801 random access analyser (Roche Diagnostics, Mannheim, Germany) [27 (link)].

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Drenthen L.C., Ajie M., Abbink E.J., Rodwell L., Thijssen D.H., Tack C.J, & de Galan B.E. (2023). No insulin degludec dose adjustment required after aerobic exercise for people with type 1 diabetes: the ADREM study. Diabetologia, 66(6), 1035-1044.

Publication 2023

Antibody insulin Assay Catecholamines Cortisol Cross reactivity Diagnostics Guinea pig Human Immunoassay method Insulin Insulin aspart Insulin degludec Insulin lispro Ms ms Plasma Polyethylene glycol Propionic anhydride Radioimmunoassay Solid phase extraction

Corresponding Organization : Radboud University Nijmegen

Top 5 similar protocols

Variable analysis

independent variables

Insulin type (aspart, lispro, degludec)

dependent variables

Plasma insulin
Plasma catecholamines
Plasma cortisol

control variables

In-house generated guinea pig anti-human insulin antibody
125I-labelled human insulin tracer
World Health Organization international standard 83/500 for assay calibration

positive controls

125I-labelled human insulin tracer generated using 125I (PerkinElmer Nederland) and human insulin (Novo Biolabs cat. no. 471)

negative controls

Not explicitly mentioned

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