Fluorescent capture assay for kinase activity

Details Fluorescent capture assay for kinase activity Fluorescent capture assay for kinase activity

G - Physics

01

N

G01N 33/53 (2006.01) A61K 31/445 (2006.01) C07K 1/13 (2006.01) C12Q 1/48 (2006.01) G01N 33/543 (2006.01) G01N 33/573 (2006.01)

Patent

CA 2388186

A method for determining the level of tyrosine kinase activity in a biological sample is disclosed. The method employs an anti-phosphotyrosine antibody as both the capture agent and the detecting agent. The detecting antibody is labeled with a fluorescent label, for instance, Cy5,Cy5.5 or Cy7 or a lanthanide ion, such as europium, as the signal generating entity. The methodis particularly well suited to high throughput screening, for example, for compounds which modulate tyrosine kinase activity.

L'invention concerne un procédé permettant de déterminer le niveau d'activité de la tyrosine kinase dans un échantillon biologique. Le procédé utilise un anticorps anti-phosphotyrosine comme agent de capture et de détection. L'anticorps de détection est marqué avec un marqueur fluorescent, Cy5,Cy5.5 ou Cy7 ou un ion lanthanide, par exemple, tel que l'europium, comme élément de signalisation. Le procédé est particulièrement recommandé pour le criblage à haut rendement, pour des composés qui modulent l'activité de la tyrosine kinase, par exemple.

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