Competitive immunoassay for haptens using protein-hapten coupled solid phase

Claim 2

Apparatus for determining progesterone in milk, comprising: (a) a solid support surface; (b) bovine serum albumin coupled to the solid support surface; and (c) progesterone covalently coupled to the bovine serum albumin.

Claim 3

Apparatus for determining hapten comprising: (a) a solid support surface; (b) a coating of albumin coupled to the solid surface; and (c) a layer of the hapten to be determined covalently coupled to the surface of the albumin.

Claim 4

Apparatus for determining hapten comprising: (a) a solid support surface; (b) a coating of albumin coupled to the solid surface; and (c) a layer of the hapten to be determined covalently coupled to the surface of the albumin.

Claim 5

In a dipstick for determining immunological species of the type which comprises an elongate stick and at least one immunogen treated pad on the stick, the improvement wherein the dipstick is suitable for determining progesterone in milk, at least one dipstick pad consisting essentially of: (a) a large surface area pad formed of pad forming constituents selected from the group consisting of fibers, particles and a combination of fibers and particles bound together to form a porous layer; (b) a coating of bovine serum albumin coupled to the pad forming constituents; and (c) progesterone covalently coupled to the bovine serum albumin.

Claim 6

In a dipstick for determining immunological species of the type which comprises an elongate stick and at least one immunogen treated pad on the stick, the improvement wherein the dipstick is suitable for determining progesterone in milk, at least one dipstick pad consisting essentially of: (a) a large surface area pad formed of pad forming constituents selected from the group consisting of fibers, particles and a combination of fibers and particles bound together to form a porous layer; (b) a coating of bovine serum albumin coupled to the pad forming constituents; and (c) progesterone covalently coupled to the bovine serum albumin.

Claim 7

The apparatus of Claim 3 further comprising: (d) a reference pad on the stick, the reference pad consisting essential of one member of a reference coupling pair covalently coupled to a large surface area pad formed of pad forming constituents selected from the group consisting of fibers, particles and a combination of fibers and particles bound together to form a porous layer, the reference coupling pair being characterized in that neither component of the reference coupling pair is reactive with either progesterone or antibodies to progesterone.

Claim 8

The apparatus of Claim 4 wherein the reference coupling pair is avidin and biotin.

Claim 9

A kit for determining progesterone in milk, comprising: (a) a dipstick comprising an elongate solid supporting stick and at least two large surface area pads on the supporting stick, the pads being characterized in that: (i) one of said pads is a reference pad consisting essentially of the large surface area pad formed of pad forming constituents selected from the group consisting of fibers, particles and a combination of fibers and particles bound together to form a porous layer, one member of a reference coupling pair bound to the pad forming constituents, the reference coupling pair being characterized in that neither member of the reference coupling pair is reactive with or interferes with the reaction of either progesterone or antibodies to progesterone; and (ii) one of said pads is a test pad consisting essentially of the large surface area pad formed of pad forming constituents selected from the group consisting of fibers, particles and a combination of fibers and particles bound together to form a porous layer, a coating of inert soluble spacer protein coupled to said pad forming constituents, and progesterone covalently coupled to the spacer protein; (b) a predetermined quantity of enzyme labelled anti-progesterone antibody; and (c) a predetermined quantity of the other member of the reference coupling pair labelled with the same enzyme with which the anti-progesterone antibody is labelled.

Claim 10

A kit for determining progesterone in milk, comprising: (a) a dipstick comprising an elongate solid supporting stick and at least two large surface area pads on the supporting stick, the pads being characterized in that: (i) one of said pads is a reference pad consisting essentially of the large surface area pad formed of pad forming constituents selected from the group consisting of fibers, particles and a combination of fibers and particles bound together to form a porous layer, one member of a reference coupling pair bound to the pad forming constituents, the reference coupling pair being characterized in that neither member of the reference coupling pair is reactive with or interferes with the reaction of either progesterone or antibodies to progesterone; and (ii) one of said pads is a test pad consisting essentially of the large surface area pad formed of pad forming constituents selected from the group consisting of fibers, particles and a combination of fibers and particles bound together to form a porous layer, a coating of inert soluble spacer protein coupled to said pad forming constituents, and progesterone covalently coupled to the spacer protein; (b) a predetermined quantity of enzyme labelled anti-progesterone antibody; and (c) a predetermined quantity of the other member of the reference coupling pair labelled with the same enzyme with which the anti-progesterone antibody is labelled.

Claim 11

The kit of Claim 6 wherein the reference coupling pair consists essentially of avidin and biotin.

Claim 12

The kit of Claim 6 further comprising developer substrate for the enzyme label.

Claim 15

The method of Claim 9 wherein the reference pad reference coupling pair consists essentially of avidin and biotin.

Claim 16

A kit for determining hapten in solution, comprising: A. a dipstick having thereon: 1. a reference pad consisting essentially of pad to which one member of a reference coupling pair is bound, the reference coupling pair being characterized in that neither member of the reference coupling pair is reactive with or interferes with the reaction of either hapten or antibodies to hapten; and 2. a test pad consisting essentially of pad having a coating of spacer protein, the spacer protein being characterized in that the protein (a) is or can be made soluble in water, (b) can be bound to the solid support, (c) can bind a hapten, and (d) does not compete with or interfere with the immunochemical reaction of the hapten in the determination; and, hapten of the type to be determined bound to the coating; B. a predetermined quantity of enzyme labelled antibody to the hapten to be determined; and C. a predetermined quantity of the other member of the reference coupling pair labelled with the same enzyme with which the anti-hapten antibody is labelled.

Claim 17

A kit for determining hapten in solution, comprising: A. a dipstick having thereon: 1. a reference pad consisting essentially of pad to which one member of a reference coupling pair is bound, the reference coupling pair being characterized in that neither member of the reference coupling pair is reactive with or interferes with the reaction of either hapten or antibodies to hapten; and 2. a test pad consisting essentially of pad having a coating of spacer protein, the spacer protein being characterized in that the protein (a) is or can be made soluble in water, (b) can be bound to the solid support, (c) can bind a hapten, and (d) does not compete with or interfere with the immunochemical reaction of the hapten in the determination; and, hapten of the type to be determined bound to the coating; B. a predetermined quantity of enzyme labelled antibody to the hapten to be determined; and C. a predetermined quantity of the other member of the reference coupling pair labelled with the same enzyme with which the anti-hapten antibody is labelled.

Claim 18

Apparatus for determining hapten comprising: (a) a solid support surface; (b) a coating of protein coupled to the solid surface; and (c) a layer of the hapten to be determined covalently coupled to the surface of the protein.

Claim 19

Apparatus for determining hapten comprising: (a) a solid support surface; (b) a coating of protein coupled to the solid surface; and (c) a layer of the hapten to be determined covalently coupled to the surface of the protein.