US 4,231,999 A · Method in assaying methods involving biospecific affinity reactions and reagent for use in said method

Abstract

A method of carrying out assaying methods involving biospecific affinity reactions in which there is used a component (I), which is labelled with at least one analytically indicatable group and is soluble in the liquid in whose presence the biospecific affinity reaction is carried out, for reaction with a counterpart (II) which exhibits biospecific affinity to (I) and optionally also one or more additional counterparts exhibiting biospecific affinity to (I) and/or (II), and forming an insoluble conjugate in which the labelled component (I) is incorporated and which conjugate is separated from the liquid phase, whereafter the analytically indicatable group is assayed in the insoluble conjugate, said method being characterized in that the analytically indicatable group is bound to the component (I) with a splittable bond of covalent nature and/or one of said counterparts is bound to a carrier, which is insoluble in said liquid, with a splittable bond of covalent nature, said splittable bonds being splittable under conditions which have practically disturbing influence on the assay method used; and in that the insoluble conjugate is split at said splittable bond or bonds, in the presence of a liquid, to form fragments which are soluble in said liquid and which contain the analytically indicatable group, which group is then assayed in the liquid phase.

Claims20

Dependent claims

In the known method of carrying out assaying methods involving biospecific affinity reactions, in which there is used a component (I) which is labelled with at least one analytically indicatable group and is soluble in the liquid in whose presence the biospecific affinity reaction is carried out, for reaction with a counterpart (II) which exhibits biospecific affinity to (I) or with said counterpart (II) and also one or more additional counterparts exhibiting biospecific affinity to at least one of (I) and (II) and forming an insoluble conjugate in which the labelled component (I) is incorporated and which conjugate is separated from the liquid phase, whereafter the analytically indicatable group is assayed in the insoluble conjugate, the improvement which comprises

(A)
(a) using as the analytically indicatable group one which is bound to the component (I) with a splittable bond of covalent nature, or
(b) using as one of said counterparts a counterpart which is bound to a carrier, which is insoluble in said liquid, with a splittable bond of covalent nature, or
(c) using as the analytically indicatable group one which is bound to the component (I) with a splittable bond of covalent nature and using as one of said counterparts a counterpart which is bound to a carrier, which is insoluble in said liquid, with a splittable bond of covalent nature, said splittable bonds, which can be the same or different, being splittable under conditions which have practically no disturbing influence on the assay method used;
and
(B) splitting the insoluble conjugate at said splittable bond or bonds, in the presence of a liquid, to form fragments which are soluble in said liquid and which contain the analytically indicatable group, which group is then assayed in the liquid phase.

3 more elements

In the known method of carrying out assaying methods involving biospecific affinity reactions, in which there is used a component (I), which is labelled with at least one analytically indicatable group and is soluble in the liquid in whose presence the biospecific affinity reaction is carried out, for reaction with a counterpart (II) which exhibits biospecific affinity to (I) or with said counterpart (II) and also one or more additional counterparts exhibiting biospecific affinity to at least one of (I) and (II) and forming an insoluble conjugate in which the labelled component (I) is incorporated and which conjugate is separated from the liquid phase, whereafter the analytically indicatable group is assayed in the insoluble conjugate, the improvement which comprises

(A)
(a) using as the analytically indicatable group one which is bound to the component (I) with a splittable bond of covalent nature, or
(b) using as one of said counterparts a counterpart which is bound to a carrier, which is insoluble in said liquid, with a splittable bond of covalent nature, or
(c) as the analytically indicatable group one which is bound to the component (I) with a splittable bond of covalent nature and as one of said counterparts a counterpart which is bound to a carrier which is insoluble in said liquid, with a splittable bond of covalent nature,
said splittable bonds, which can be the same or different, being splittable under conditions which have practically no disturbing influence on the assay method used and said labelled component (I) comprising a multi-conjugate of a plurality of analytically indicatable groups linked together via linkages containing splittable bonds, said multiconjugate of indicatable groups in turn being bound to the component (I);
and
(B) splitting the insoluble conjugate at said splittable bond or bonds, in the presence of a liquid, to form fragments which are soluble in said liquid and which contain the analytically indicatable group, which group is then assayed in the liquid phase.

4 more elements

Dependencies are inferred from the claim text.

Method in assaying methods involving biospecific affinity reactions and reagent for use in said method

Life of this patent

Method in assaying methods involving biospecific affinity reactions and reagent for use in said method

Life of this patent

Abstract

Claims20