US4481298A — Pre-precipitated double antibody immunoassay meth…

Claim 1

Independent Claim 149 words

An immunoassay process for the detection of antigen in a sample, which comprises: a. forming a mixture of said sample in an appropriate medium therefor with 1. a preformed antigen-binding complex of a primary antibody having specificity for said antigen and containing a F c region, and a secondary antibody having specificity only for F c region of said primary antibody; wherein said secondary antibody is affinity purified; and with 2. A detectably labeled form of said antigen; b. incubating said mixture formed in step (a), which comprises said complex suspended in said medium, for a time sufficient to allow said antigen and said detectably labeled antigen to competitively bind to the primary antibody of said preformed complex; c. Separating said complex containing bound labeled antigen from said suspension medium; and d. detecting said separated complex containing bound labeled antigen, or said separated suspension medium containing free labeled antigen.

Claim 2

Dependent Claim 149 words

An immunoassay process for the detection of antigen in a sample, which comprises: a. forming a mixture of said sample in an appropriate medium therefor with 1. a preformed antigen-binding complex of a primary antibody having specificity for said antigen and containing a F c region, and a secondary antibody having specificity only for F c region of said primary antibody; wherein said secondary antibody is affinity purified; and with 2. A detectably labeled form of said antigen; b. incubating said mixture formed in step (a), which comprises said complex suspended in said medium, for a time sufficient to allow said antigen and said detectably labeled antigen to competitively bind to the primary antibody of said preformed complex; c. Separating said complex containing bound labeled antigen from said suspension medium; and d. detecting said separated complex containing bound labeled antigen, or said separated suspension medium containing free labeled antigen.

Claim 3

Dependent Claim 13 words

The process of claim 1 wherein said secondary binding macromolecule is an antibody.

Claim 4

Dependent Claim 20 words

The process of claims 1 or 2 wherein said preformed complex is free of any polymeric insoluble solid phase matrix.

Claim 5

Dependent Claim 17 words

The process of claim 1 wherein said primary antibody is present in an amount of 1-40 μg/ml.

Claim 6

Dependent Claim 17 words

The process of claim 4 wherein said primary antibody is present in an amount of 1-10 μg/ml.

Claim 7

Dependent Claim 24 words

The process of claim 1 wherein said preformed complex is free of non-specific IgG belonging to the same animal species as said primary antibody.

Claim 8

Dependent Claim 26 words

The process of claim 1 wherein said secondary binding macromolecule has been affinity purified by contact with an insoluble solid phase having Fc fragments attached thereon.

Claim 9

Dependent Claim 21 words

The process of claim 2 wherein said antibody has been raised in an animal by immunizing said animal with Fc fragments.

Claim 10

Dependent Claim 37 words

The process of claim 1 wherein said secondary binding macromolecule has been affinity purified by sequential contact with an insoluble solid phase having Fc fragments attached thereon, and with another solid phase having human IgG attached thereon.

Claim 11

Dependent Claim 19 words

The process of claim 1 wherein said detectably labeled antigen is labeled with a radiolabel or an enzyme label.

Claim 12

Dependent Claim 12 words

The process of claim 1 wherein said antigen is T 3 .

Claim 13

Dependent Claim 12 words

The process of claim 1 wherein said antigen is T 4 .

Claim 14

Dependent Claim 10 words

The process of claim 1 wherein said antigen is digoxin.

Claim 15

Dependent Claim 38 words

An antigen-binding complex of an antigen-binding primary antibody containing a F c region unmodified, and a secondary unmodified antibody having specificity only for the F c region of the primary, antibody, wherein said secondary antibody is affinity purified.

Claim 16

Dependent Claim 38 words

An antigen-binding complex of an antigen-binding primary antibody containing a F c region unmodified, and a secondary unmodified antibody having specificity only for the F c region of the primary, antibody, wherein said secondary antibody is affinity purified.

Claim 17

Dependent Claim 17 words

The complex of any of claims 14 which is free from any polymeric, insoluble solid phase matrix.

Claim 18

Dependent Claim 20 words

The complex of claim 14 which is free of non-specific IgG belonging to the same animal species as said antibody.

Claim 19

Dependent Claim 26 words

The complex of claim 14 wherein said secondary antibody has been affinity purified by contact with an insoluble solid phase having F c fragments attached thereon.

Claim 20

Dependent Claim 23 words

The complex of claim 14 wherein said second antibody has been raised in an animal by immunizing said animal with F c fragments.

Claim 21

Dependent Claim 36 words

The complex of claim 14 wherein said binding macromolecule has been affinity purified by sequential contact with an insoluble solid phase having Fc fragments attached thereon, and with another solid phase having human IgG attached thereon.

Claim 22

Dependent Claim 91 words

A kit comprising a carrier being compartmentalized to receive at least two container means therein wherein the first of said container means comprises a preformed antigen binding complex of a primary antibody having specificity for said antigen, and having a F c region, and a secondary unmodified antibody having specificity only for the F c region of said primary antibody, wherein said secondary antibody is affinity purified, the second of said container means comprises a detectably labeled form of an antigen having affinity towards said primary antibody in said preformed complex.

Claim 23

Dependent Claim 91 words

A kit comprising a carrier being compartmentalized to receive at least two container means therein wherein the first of said container means comprises a preformed antigen binding complex of a primary antibody having specificity for said antigen, and having a F c region, and a secondary unmodified antibody having specificity only for the F c region of said primary antibody, wherein said secondary antibody is affinity purified, the second of said container means comprises a detectably labeled form of an antigen having affinity towards said primary antibody in said preformed complex.

Claim 24

Dependent Claim 19 words

The kit of claim 20 wherein said preformed complex in said first container means is dissolved in a buffer.

Claim 25

Dependent Claim 17 words

The kit of claim 20 wherein said detectably labeled antigen in said second container means is lyophilized.

Claim 26

Dependent Claim 20 words

The kit of claim 20 wherein said detectably labeled antigen in said second container means is dissolved in a buffer.

Claim 27

Dependent Claim 44 words

The kit of claim 20 which additionally contains a plurality of container means each comprising a mixture of said detectably labeled antigen and a non-labeled form of said antigen, each of said mixtures having a different molar ratio of detectably labeled to non-labeled antigen.

Claim 28

Dependent Claim 30 words

The kit of any of claims 20, 21, 22, 23, or 24 wherein said preformed complex in said first container means is free of any insoluble polymeric solid phase matrix.

Claim 29

Dependent Claim 36 words

The kit of any of claims 20, 21, 22, 23, or 24 wherein said preformed complex in said first container means is free of non-specific IgG belonging to the same animal species as said primary antibody.

Claim 30

Dependent Claim 32 words

The kit of any of claims 20, 21, 22, 23, or 24 wherein said secondary antibody has been purified by contact with an insoluble solid phase having F c fragments attached thereon.

Claim 31

Dependent Claim 22 words

The kit of any of claims 20, 21, 22, 23, or 24 wherein said detectably labeled antigen is 125 I-T 3 .

Claim 32

Dependent Claim 22 words

The kit of any of claims 20, 21, 22, 23, or 24 wherein said detectably labeled antigen is 125 I-T 4 .

Claim 33

Dependent Claim 20 words

The kit of any of claims 20, 21, 22, 23, or 24 wherein said detectably labeled antigen is 125 I-digoxin.

Claim 34

Dependent Claim 28 words

The kit of claim 28 wherein said secondary antibody in said first container means has been rasied in an animal by immunizing said animal with F c fragments.

Claim 35

Dependent Claim 26 words

The kit of any of claims 20, 21, 22, 23, or 24 wherein said detectably labeled antigen is labeled with a radiolabel or an enzyme label.

Claim 36

Dependent Claim 83 words

A kit comprising a carrier being compartmentalized to receive at least one container means wherein said container means comprises (1) a preformed antigen-binding complex of a primary antibody having specificity for said antigen, and containing a F c region, secondary unmodified antibody having specificity only for the F c region of said primary antibody, wherein said secondary antibody is affinity purified, and (2) a predetermined amount of a detectably labeled form of an antigen having affinity towards the primary antibody in said complex.

Claim 37

Dependent Claim 83 words

A kit comprising a carrier being compartmentalized to receive at least one container means wherein said container means comprises (1) a preformed antigen-binding complex of a primary antibody having specificity for said antigen, and containing a F c region, secondary unmodified antibody having specificity only for the F c region of said primary antibody, wherein said secondary antibody is affinity purified, and (2) a predetermined amount of a detectably labeled form of an antigen having affinity towards the primary antibody in said complex.

Claim 38

Dependent Claim 168 words

An immunoassay process for the detection of Thyroxine Binding Globulin (TBG) in a sample, which comprises: a. forming a mixture of said sample in an appropriate medium therefor with 1. A preformed antigen-binding complex of a primary antibody, having specificity for T 3 and containing a F c region, and a secondary antibody having specificity only for the F c region of said primary antibody; wherein said secondary antibody is affinity purified; and with 2. a detectably labeled form of T 3 ; b. incubating said mixture formed in step (a), which comprises said complex suspended in said medium, for a time sufficient to allow said antigen and said detectably labeled T 3 to competitively bind to the primary antibody of said preformed complex and to said TBG in said sample; c. separating said complex containing bound labeled T 3 from said suspension medium; and d. detecting said separated complex containing bound labeled T 3 , or said separated suspension medium containing labeled T 3 bound to TBG.

Claim 39

Dependent Claim 168 words

An immunoassay process for the detection of Thyroxine Binding Globulin (TBG) in a sample, which comprises: a. forming a mixture of said sample in an appropriate medium therefor with 1. A preformed antigen-binding complex of a primary antibody, having specificity for T 3 and containing a F c region, and a secondary antibody having specificity only for the F c region of said primary antibody; wherein said secondary antibody is affinity purified; and with 2. a detectably labeled form of T 3 ; b. incubating said mixture formed in step (a), which comprises said complex suspended in said medium, for a time sufficient to allow said antigen and said detectably labeled T 3 to competitively bind to the primary antibody of said preformed complex and to said TBG in said sample; c. separating said complex containing bound labeled T 3 from said suspension medium; and d. detecting said separated complex containing bound labeled T 3 , or said separated suspension medium containing labeled T 3 bound to TBG.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1987-11-06	1988-05-07	1988-11-06	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1991-11-06	1992-05-07	1992-11-06	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	1995-11-06	1996-05-07	1996-11-06	$7700.00	Window Closed / Lapsed

Pre-precipitated double antibody immunoassay method

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