Enzyme amplified competitive and sandwich chelation assays...

G - Physics – 01 – N

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Details

G01N 33/20 (2006.01) G01N 33/58 (2006.01) G01N 33/84 (2006.01)

Patent

CA 2172620

The present invention provides methods for assaying for a metal ion in a sample. In one aspect, the present invention relates to an enzyme amplified sandwich assay. This method relies upon the ability of the metal ion to form a complex with two chelators. Such an association is termed a sandwich complex with the metal ion forming the middle of the sandwich. In this method the first chelator is immobilized on a solid support, while the second chelator is linked to a reporter group. This arrangement forms a highly selective, sensitive, and convenient system for the quantitative, detection of metal ions. This approach combines the specific recognition of the metal ion by the first and second chelators, with the great signal amplification offered by enzymes or other reporter groups. In another aspect, the present invention relates to a competitive assay that relies on the competitive inhibition of complex formation between a chelator immobilized on a solid support and an organometallic compound attached to a reporter group, by the metal ions of interest in a sample.

Procédés de dosage d'un ion métal dans un échantillon. Selon un aspect, cette invention permet un dosage du type en sandwich à amplification enzymatique. Ce procédé se fonde sur la capacité de l'ion métal à former un complexe avec deux chelateurs, une telle association étant appelée un complexe en sandwich dans lequel l'ion métal constitue le milieu du sandwich. Selon ce procédé le premier chélateur est immobilisé sur un support solide alors que le deuxième chélateur est lié à un groupe reporter. Cette configuration forme un système hautement sélectif, sensible et commode pour détecter quantitativement les ions métal. Cette approche combine la reconnaissance spécifique de l'ion métal par les premier et deuxième chélateurs et l'amplification sensible du signal offerte par les enzymes ou d'autres groupes reporters. Selon un autre aspect, cette invention concerne un dosage compétitif qui se fonde sur l'inhibition compétitive de la formation de complexe entre un chélateur immobilisé sur un support solide et un composé organométallique attaché à un groupe reporter, par les ions métal recherchés dans un échantillon.

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