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A homogeneous receptor-ligand assay based on fluorescence resonance energy transfer is described. In the assay, recombinant human interleukin 2 (IL-2) and a monoclonal antibody against the human IL-2 receptor alpha chain were labelled with a highly fluorescent europium chelate and Cy5, respectively. As a result of a successful receptor-ligand complex formation, these labels are brought into close proximity, which will thereby allow an energy transfer to occur from the donor (europium) to the acceptor (Cy5), upon excitation of the donor. Utilization of specific non-neutralizing antibodies made it possible to use crude hIL-2R alpha membranes prepared from recombinant baculovirus-infected Sf9 insect cells. The specific energy transfer was measured at the emission wavelength of Cy5 using a time-resolved fluorometer. The data presented, demonstrate that this assay design can be utilized for saturation as well as competitive binding experiments in addition to regular receptor titrations. As a rapid simple homogeneous assay it is particularily suitable for high throughput screening analyses.
K Stenroos, and
P Hurskainen, and
S Eriksson, and
I Hemmilä, and
K Blomberg, and
C Lindqvist
April 2006,
Analytical biochemistry,
K Stenroos, and
P Hurskainen, and
S Eriksson, and
I Hemmilä, and
K Blomberg, and
C Lindqvist
March 2005,
Analytical chemistry,
K Stenroos, and
P Hurskainen, and
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K Blomberg, and
C Lindqvist
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C Lindqvist
June 2008,
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K Stenroos, and
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K Stenroos, and
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K Stenroos, and
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C Lindqvist
July 2001,
Nucleic acids research,
K Stenroos, and
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C Lindqvist
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Journal of biomolecular screening,
K Stenroos, and
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I Hemmilä, and
K Blomberg, and
C Lindqvist
May 2018,
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K Stenroos, and
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I Hemmilä, and
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C Lindqvist
December 2001,
Talanta,
