Solid phase IL-2-IL-2R alpha assay with time resolved fluorometry.

K Stenroos; P Hurskainen; K Blomberg; C Lindqvist

doi:10.1016/s0165-2478(97)02719-3

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PubMed · 9436463

Solid phase IL-2-IL-2R alpha assay with time resolved fluorometry.

K Stenroos·P Hurskainen·K Blomberg·C Lindqvist

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Abstract

A time-resolved fluorometric, solid phase, receptor ligand interaction assay is described. The assay consists of wells coated with anti-human IL-2 receptor alpha (hIL-2R alpha) monoclonal antibodies (mAb), europium labelled hIL2 (Eu-IL-2) and human recombinant IL-2 receptor alpha subunits expressed in the baculovirus expression vector system (BEVS). In the assay hIL-2R alpha-Eu-IL-2 complexes bind to the solid phase mAb. Receptor bound Eu is dissociated into an enhancer solution where it forms highly fluorescent complexes. The fluorescence is measured in a time-resolved fluorometer. The Kd value calculated from the saturation curve is in good agreement with previously reported values for the low affinity type of IL-2R, making the described assay a simple and nonradioactive alternative for measurement of soluble hIL-2R alpha in biological systems. Furthermore this assay format provides convenient separation of bound ligand from unbound and is therefore suitable for high throughput screenings.

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Europium Fluorescent Dyes Fluorometry Humans Immunosorbent Techniques Interleukin-2 Ligands Protein Binding Receptors, Interleukin-2 Recombinant Proteins Journal Article Research Support, Non-U.S. Gov't

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K Stenroos, P Hurskainen, K Blomberg, C Lindqvist. 1997. Solid phase IL-2-IL-2R alpha assay with time resolved fluorometry.. https://doi.org/10.1016/s0165-2478(97)02719-3

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