Biospecific binding reactants labeled with new luminescent lanthanide chelates and their use

A detectable molecule which comprises a biospecific binding reactant attached to a luminescent lanthanide chelate comprising a lanthanide ion and a chelating ligand of formula (I)

• 10
• wherein
• R1 is selected from the group consisting of H, —COOH, —COO−, —CH2COOH and —CH2COO−;
• G1 is a group consisting of one or two moieties, each moiety being selected from the group consisting of ethynediyl (—C≡C—), ethenylene (—CH═CH—), phenylene, biphenylene, naphthylene, pyridylene, pyrazinylene, pyrimidinylene, pyridazinylene, furylene, thienylene, pyrrolylene, imidazolylene, pyrazolylene, thiazolylene, isothiazolylene, oxazolylene, isoxazolylene, fyrazanylene, 1,2,4-triazol-3,5-ylene and oxadiazolylene;
• G2 for coupling to a biospecific binding reactant is selected from the group consisting of thiourea (—NH—CS—NH—), aminoacetamide (—NH—CO—CH2—NH—), amide (—NH—CO—, —CO—NH—, —NCH3—CO— and —CO—NCH3—), aliphatic thioether (—S—), disulfide (—S—S—) and 6-substituted-1,3,5-triazine-2,4-diamine;
• Z is selected from the group consisting of carboxyalkyl amine {—N[(CH2)nCOOH]— or —N[(CH2)nCOO−]— wherein n=1, 2, 3, 4, 5 or 6}, ether (—O—), thioether (—S—), carbonyl (—CO—) and unsubstituted or substitute methyl (—CR2—) wherein group R2 is selected from the group consisting of H, methyl, ethyl and carboxylalkyl [—(CH2)nCOOH or —(CH2)nCOO− wherein n=1, 2, 3, 4, 5 or 6];
• and the lanthanide ion is europium(III), terbium(III), dysprosium(III) or samarium(III).

A method of carrying out a biospecific binding assay using a detectable molecule to an analyte to be determined, said detectable molecule comprising a biospecific binding reactant attached to a luminescent lanthanide chelate which comprises a lanthanide ion and a chelating ligand of formula (I)

• 11
• wherein
• R1 is selected from the group consisting of H, —COOH, —COO−, —CH2COOH and —CH2COO−;
• G1 is a group consisting of one or two moieties, each moiety being selected from the group consisting of ethynediyl (—C≡C—), ethenylene (—CH═CH—), phenylene, biphenylene, naphthylene, pyridylene, pyrazinylene, pyrimidinylene, pyridazinylene, furylene, thienylene, pyrrolylene, imidazolylene, pyrazolylene, thiazolylene, isothiazolylene, oxazolylene, isoxazolylene, fyrazanylene, 1,2,4-triazol-3,5-ylene and oxadiazolylene;
• G2 for coupling to a biospecific binding reactant is selected from the group consisting of thiourea (—NH—CS—NH—), aminoacetamide (—NH—CO—CH2—NH—), amide (—NH—CO—, —CO—NH—, —NCH3—CO— and —CO—NCH3—), aliphatic thioether (—S—), disulfide (—S—S—) and 6-substituted-1,3,5-triazine-2,4diamine;
• Z is selected from the group consisting of carboxyalkyl amine {—N[(CH2)nCOOH]— or —N[(CH2)nCOO−]— wherein n=1, 2, 3, 4, 5 or 6}, ether (—O—), thioether (—S—), carbonyl (—CO—) and unsubstituted or substitute methyl (—CR2—) wherein group R2 is selected from the group consisting of H, methyl, ethyl and carboxylalkyl [—(CH2)nCOOH or —(CH2)nCOO− wherein n=1, 2, 3, 4, 5 or 6];
• and the lanthanide ion is europium(III), terbium(III), dysprosium(III) or samarium(III).