Enzyme-free diagnostic binding reagents
First Claim
1. An enzyme-free diagnostic reagent for determining the presence of an entity to be assayed in a homogeneous assay comprising an enzyme-free hydrolytic catalyst coupled by a linking group to a first binding agent, said catalyst having a molecular weight together with the linking group of about 75 to about 2000 daltons, wherein said catalyst catalyzes a hydrolytic chemical reaction with successive preselected hydrolyzable co-reactant molecules when said co-reactant molecules and said catalyst are admixed in homogenous solution and said catalyst is coupled to said first binding agent as said diagnostic reagent;
- said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entity needed for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed.
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Abstract
An enzyme-free diagnostic reagent, methods of its use and diagnostic systems containing that reagent are disclosed. The enzyme-free diagnostic reagent comprises an enzyme-free catalyst coupled by a linking group to a first binding agent, binds in aqueous medium to a second binding moiety to form a binding complex, and indicates the amount of second binding moiety present in the complex by means of its catalytic reactivity with co-reactant molecules.
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Citations
42 Claims
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1. An enzyme-free diagnostic reagent for determining the presence of an entity to be assayed in a homogeneous assay comprising an enzyme-free hydrolytic catalyst coupled by a linking group to a first binding agent, said catalyst having a molecular weight together with the linking group of about 75 to about 2000 daltons, wherein said catalyst catalyzes a hydrolytic chemical reaction with successive preselected hydrolyzable co-reactant molecules when said co-reactant molecules and said catalyst are admixed in homogenous solution and said catalyst is coupled to said first binding agent as said diagnostic reagent;
- said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entity needed for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16)
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17. A diagnostic kit comprising at least one package containing an aqueous solution including a known amount of an enzyme-free diagnostic reagent for determining the presence of an entity to be assayed in a homogeneous assay, said diagnostic reagent comprising an enzyme-free hydrolytic catalyst coupled by a linking group to a first binding agent, said catalyst having a molecular weight together with the linking group of about 75 to about 2000 daltons, wherein said catalyst catalyzes a hydrolytic chemical reaction with successive preselected hyrolyzable co-reactant molecules when said co-reactant molecules and said catalyst are admixed in homogeneous solution and said catalyst is coupled to said first binding agent as said diagnostic reagent;
- said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entitity needed for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed.
- View Dependent Claims (18)
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19. A method of carrying out a binding assay comprising the steps of
(a) providing an enzyme-free diagnostic reagent for determining the presence of an entity to be assayed in a homogeneous assay comprising an enzyme-free hydrolytic catalyst coupled by a linking group to a first binding agent, said catalyst having a molecular weight together with the linking group of about 75 to about 2000 daltons, wherein said catalyst catalyzes a hydrolytic chemical reaction with successive preselected hydrolyzable co-reactant molecules when said co-reactant molecules and said catalyst are admixed. in homogeneous solution and said catalyst is coupled to said first binding agent as said diagnostic reagent; - said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entitity needed for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed;
(b) providing a first aqueous composition containing said second binding moiety; (c) admixing in an aqueous medium a known, predetermined amount of said diagnostic reagent and a known, predetermined amount of said first aqueous composition to form a binding complex between said diagnostic reagent and said second binding moiety; (d) admixing a plurality of hydrolyzable co-reactant molecules with the admixture of step (c); and (e) measuring the reaction between said enzyme-free catalyst and said co-reactant molecules. - View Dependent Claims (20, 21, 22, 23, 24, 25, 26, 27)
- said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entitity needed for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed;
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28. An enzyme-free diagnostic reagent for determining the presence of an entity to be assayed in a homogeneous assay comprising an enzyme-free hydrolytic catalyst coupled by a linking group to a first binding agent, said enzyme-free catalyst prior to being coupled to said first binding agent having a structure corresponding to a formula selected from the group consisting of ##STR11## wherein R1 is hydrogen, C1 -C12 alkyl or Z, or R1 and Z taken together form a 5- or 6-membered ring that contains a linking moiety;
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R2 is hydrogen or C1 -C4 alkyl; R3 is C1 -C4 alkyl, and R4 is alkyl or substituted arylalkyl containing 1 to about 18 carbon atoms, or two of R3, R4 and Z taken together form a 5- or 6- membered ring containing a linking moiety, or R3, R4 and Z taken together form a bicyclic ring compound containing a linking moiety and wherein each of the rings of the bicyclic ring compound contains 5- or 6-membered ring that contains a linking moiety; and Z is a linking group including a linking moiety prior to linking of the catalyst to the first binding agent; said catalyst having a molecular weight together with the linking group of about 75 to about 2000 daltons, wherein said catalyst catalyzes a hydrolytic chemical reaction with successive preselected hydrolyzable co-reactant molecules when said co-reactant molecules and said catalyst are admixed in homogeneous solution and said catalyst is coupled to said first binding agent as said diagnostic reagent;
said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entity needed for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed. - View Dependent Claims (29, 30, 31, 32)
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33. A diagnostic kit comprising at least one package containing an aqueous solution including a known amount of an enzyme-free diagnostic reagent for determining the presence of an entity to be assayed in a homogeneous assay, said diagnostic reagent comprising an enzyme-free hydrolytic catalyst coupled by a linking group to a first binding agent, said enzyme-free hydrolytic catalyst prior to being coupled to said first binding agent has a structure corresponding to a formula selected from the group consisting of ##STR12## wherein R1 is hydrogen, C1 -C12 alkyl or Z, or R1 and Z taken together form a 5- or 6- members;
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R2 is hydrogen or C.sub. -C4 alkyl; R3 is C1 -C4 alkyl, and R4 is alkyl or substituted arylalkyl containing 1 to about 18 carbon atoms, or two of R3, R4 and Z taken together form a 5- or 6- membered ring containing a linking moiety, or R3, R4 and Z taken together form a bicyclic ring compound containing a linking moiety and wherein each of the rings of the bicyclic ring compounds contains 5- or 6-membered ring that contains a linking moiety; and Z is a linking group including a linking moiety prior to linking of the catalyst to the first binding agent; said catalyst having a molecular weight together with the linking group of about 75 to about 2000 daltons, wherein said catalyst catalyzes a hydrolytic chemical reaction with successive preselected hydrolyzable co-reactant molecules when said co-reactant molecules and said catalyst are admixed in homogeneous solution and said catalyst is coupled to said first binding agent as said diagnostic reagent;
said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entity for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed. - View Dependent Claims (34, 35, 36, 37)
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38. A method of carrying out a binding assay comprising the steps of
(a) providing an enzyme-free diagnostic reagent for determining the presence of an entity to be assayed in a homogeneous assay comprising an enzyme-free hydrolytic catalyst coupled by a linking group to a first binding agent, said enzyme-free hydrolytic catalyst prior to being coupled to said first binding agent has a structure corresponding to a formula selected from the group consisting of ##STR13## wherein R1 is hydrogen, C1 -C12 alkyl or Z, or R1 and Z taken form a 5- or 6-membered ring that contains a linking moiety; -
R2 is hydrogen or C1 -C4 alkyl; R3 is C.sub. -C4 alkyl, and R4 is alkyl or substituted arylalkyl containing 1 to about 18 carbon atoms, or two of R3, R4 and Z taken together form a 5- or 6-membered ring containing a linking moiety, or R3, R4 and Z taken together form a bicyclic ring compound containing a linking moiety and wherein each of the rings of the bicyclic ring compound contains 5- or 6-membered ring that contains a linking moiety; and Z is a linking group including a linking moiety prior to linking of the catalyst to the first binding agent. said cataylst having a molecular weight together with the linking group of about 75 to about 2000 daltons, wherein said catalyst catalyzes a hydrolytic chemical reaction with successive preselected hydrolyzable co-reactant molecules when said co-reactant molecules and said catalyst are admixed in homogeneous solution and said catalyst is coupled to said first binding agent as said diagnostic reagent;
said diagnostic reagent specifically binding in aqueous medium to a second binding moiety to form a binding complex, said second binding moiety being (a) the entity to be assayed in a direct assay, or (b) an entity needed for the assay in an indirect assay, said diagnostic reagent indicating the amount of said second binding moiety present in said binding complex by means of the catalytic reactivity of said catalyst with said co-reactant molecules, said catalytic reactivity being changed when said binding complex is formed;(b) providing a first aqueous composition containing said second binding moiety; (c) admixing in an aqueous medium a known, predetermined amount of said diagnostic reagent and a known, predetermined amount of said first aqueous composition to form a binding complex between said diagnostic reagent and said second binding moiety; (d) admixing a plurality of hydrolyzable co-reactant molecules with the admixture of step (c); and (e) measuring the reaction between said enzyme-free catalyst and said co-reactant molecules. - View Dependent Claims (39, 40, 41, 42)
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Specification