Wallac Oy

Radiometer Turku Oy

Innotrac Diagnostics Oy

Radiometer Medical Aps

Abacus Diagnostica Oy

Dhr Finland Oy

Harri Takalo

Veli-Matti Mukkala

Jouko Kankare

Jari Hovinen

Niko Meltola

Elina Hanninen

Matti Helenius

Henri Sund

Kim Pettersson

Ilkka Hemmila

Kaj Blomberg

Heikki Mikola

Päivi Liitti

Qiu-Ping Qin

Allan Milton Byrnard

Johanna Ylikotila

Kirsten Marie Jakobsen

Lasse Välimaa

Juan Carlos Rodriguez-Ubis

Elina Tienari

Jaana Rosenberg

Johann Ylikotila

The present invention relates to a background blocking concept for use in time-resolved fluorometry binding assays. More particular, the invention relates to a binding assay and a kit involving the use of the same or similar chelating ligand in lanthanide chelate-labelled analyte-specific biomolecules and as or in a background blocking agent.

Background blockers for binding assays

The present application discloses novel lanthanide chelate designs (Formula (I) and Formula (III)) having fluorenyl-, fluorenylethynyl, 9H-carbazolyl-, 9H-carbozolylethynyl-, dibenzothiophenyl-, dibenzothiophenylethynyl-, dibenzofuranyl or dibenzofuranylethynyl pyridine chromophores around an emitting lanthanide ion, e.g. an europium(III) ion. The three-membered ring chromophores exhibit high molar absorptivity and luminescence with lanthanide ions. The application also discloses a detectable molecule comprising a biospecific binding reactant useful in bioaffinity based binding assay, luminescent lanthanide chelating ligands, as well as a solid support conjugated with the chelates. 

Chromophoric structures for lanthanide chelates

The present application discloses a luminescent lanthanide chelate of formula (I) with lanthanides such as europium, as well as the corresponding luminescence lanthanide chelating ligand. The application also discloses a detectable molecule comprising a biospecific binding reactant (such as an antibody) conjugated to the luminescent lanthanide chelate as well as a method of carrying out a biospecific binding assay, the use of such a detectable molecule in a specific bioaffinity based assay, the use of such a detectable molecule in a specific bioaffinity based binding assay utilizing time-resolved fluorometric determination of a specific luminescence, and a solid support material conjugated with the luminescent lanthanide chelate.

Luminescent lanthanide chelates having three chromophores an their use

The present application discloses a luminescent lanthanide chelate comprising one or more chromophoric moieties of the formula (I) or of the formula (III)  wherein R, Rand Reach independently are selected from carbon-containing substituents forming a C&#x2014;O bond with the neighbouring oxygen atom, Rand Reach represent a bond between the chromophoric moiety and other moieties of the chelate, and Lnis a lanthanide ion, as well as the corresponding luminescence lanthanide chelating ligand. The application also discloses a detectable molecule comprising a biospecific binding reactant (such as an antibody) conjugated to the luminescent lanthanide chelate as well as a method of carrying out a biospecific binding assay, the use of such a detectable molecule in a specific bioaffinity based binding assay utilizing time-resolved fluorometric determination of a specific luminescence, and a solid support material conjugated with the luminescent lanthanide chelate.

Luminescent lanthanide chelates with enhanced excitation properties

A high capacity solid phase for bioaffinity assays and other solid phase applications prepared by coating a solid support with an analyte-specific biomolecule in the presence of a zwitterionic additive is described. Structures prepared according to the invention provide high capacity solid phases with enhanced binding properties, which are advantageous for any solid phase based assay.

High capacity solid phase

Substrates for thrombin and assays for determining the level of bioactive thrombin in a sample are disclosed, wherein the substrate has the general formula: A-X&#x2014;Z-A' wherein one of either A or A&#x2032; comprises a luminescent chelate and the other one of A or A&#x2032; comprises a first partner of a binding pair, X forms a tri- or tetra-peptide, and Z comprises a linker.

Thrombin substrate and assay for determining the level of bioactive thrombin in a sample

The invention relates to a detectable molecule comprising a biospecific binding reactant attached to a luminescent lanthanide chelate comprising a lanthanide ion and a chelating ligand of the formula (I) wherein, RRare independently of each other selected from the group consisting of hydrogen, methyl, ethyl, &#x2014;COOH, &#x2014;COO&#x2014;, &#x2014;CHCOOH, &#x2014;CHCOO&#x2014;, hydroxyl or OR; Ris selected from the group consisting of &#x2014;CH, &#x2014;C(CH), &#x2014;C(CR), wherein Ris an alkyl with 1 to 6 carbon atoms, &#x2014;CHCOOH, &#x2014;CHCOO, and appropriate hexose residues; Ris a linker for coupling to a biospecific binding reactant selected from the group consisting of thiourea (&#x2014;NH&#x2014;CS&#x2014;NH&#x2014;), aminoacetamide (&#x2014;NH&#x2014;CO&#x2014;CH&#x2014;NH&#x2014;), amide (&#x2014;NH&#x2014;CO&#x2014; and &#x2014;CO&#x2014;NH&#x2014;), aliphatic thioether (&#x2014;S&#x2014;), disulfide (&#x2014;S&#x2014;S&#x2014;) and 6-substituted-1,3,5-triazine-2,4-diamine; and the lanthanide ion is selected from the group consisting of europium(III), terbium(III), dysprosium(III) and samarium(III). The invention also relates to corresponding lanthanide chelates. The invention further relates to the use of the detectable molecule of the invention.

Luminescent lanthanide labelling reagents and their use

This invention relates to a luminescent lanthanide chelate comprising a lanthanide ion and a chelating ligand of formula (I)wherein Ris selected from the group consisting H, &#x2014;COOH, &#x2014;COO, &#x2014;CHCOOH and &#x2014;CHCOO; Gis a group consisting of one or two moieties each moiety being selected from the group consisting of ethynediyl, ethenylene, 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; Gfor coupling to a biospecific binding reactant is selected from the group consisting of amino, aminooxy, carbonyl, aldehyde or mercapto groups and activated forms made of them; Z is selected from the group consisting of carboxyalkyl amine, ether, thioether, carbonyl and unsubstituted or substitute methyl (&#x2014;CR&#x2014;) wherein group Ris selected from the group consisting of H, methyl, ethyl and carboxylalkyl; and the lanthanide ion is europium(III), terbium(III), dysprosiym(III) or samarium(III). This invention further relates to a detectable molecule comprising the lanthanide chelate and the use of the molecule in a method of carrying out a biospecific binding assay.

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

The invention relates to a novel labeling reactant of formula (I) suitable for labeling an oligonucleotidewherein:R is a temporary protecting group. A is either a phosphorylating moiety or a solid support tethered to a bridge point Z via a linker arm E. E' is a linker arm between G and Z. G is a bivalent aromatic structure, tethered to two iminodiacetic acid ester groups N(COOR&#x2032;&#x2033;)or G is a structure selected from a group consisting ofG is a protected functional group. The invention further concerns a method for direct attachment of a conjugate group to an oligonucleotide structure enabling the attachment of a desired number of these groups during chain assembly. The method comprises a Mitsonobu alkylation.

Oligonucleotide labeling reactants and their use

The invention relates to a novel labeling reactant, suitable for labeling of a biospecific binding reactant using solid-phase synthesis. The invention further concerns new labeling methods. The novel labeling reactant has the formula (I) ##STR1## wherein -A- is a bivalent aromatic structure capable of absorbing light or energy and transferring the excitation energy to a lanthanide ion after the product made by the said solid-phase synthesis has been released from the used solid support, deprotected and converted to a lanthanide chelate;

Labeling reactants and their use

This invention relates to a detectable molecule comprising a biospecific binding reactant attached to a luminescent lanthanide chelate comprising a lanthanide ion and a chelating ligand of the formula ##STR1## wherein --A-- is a bivalent aromatic structure selected from the group consisting of ##STR2## One of the two groups G.sub.1 or G.sub.2 is used for coupling the chelate to a biospecific binding reactant. The lanthanide ion is europium(III), terbium(III), dysprosium (III) or samarium (III).

Biospecific binding reactants labelled with luminescent lanthanide

Luminescent lanthanide chelates

A method of conducting assays for analytes, usually from biological samples, utilizing bioaffinity reagents linked to luminescent lanthanide chelates, which are synthesized so that the non-radiative quenching of the ion luminescence through C--H bond vibrational energy manifolds is avoided using a stable chelate where the CH and CH.sub.2 groups in the vicinity of the emittive ion are substituted with CD and CD.sub.2 groups hence producing improved luminescence quantum yield and higher assay sensitivities.

Luminescent lanthanide chelates with decreased non-radiative energy loss

The invention pertains to terpyridine compounds having structure (I). These compounds form fluorescent lanthanide chelates with the appropriate metal ions. The fluorescent metal chelates are useful as probes in time-resolved fluorescence spectroscopy.

Growing community of inventors

Turku, Finland

Harri Takalo