Immunochemical process and reagent for the determination of a polyvalent antigen in a liquid sample
First Claim
1. Process for the quantitative determination of a polyvalent antigen comprising incubating a liquid sample containing said antigen with three different receptors, wherein the first receptor (R1) and the third receptor (R3) are in liquid phase and bind with the polyvalent antigen, and the second receptor (R2) is in solid phase and binds with (R1), wherein (R3) carries a label and does not cross react with (R1) and (R2) to form a solid phase complex of (R2), (R1), antigen and (R3), separating the solid phase from the liquid phase and measuring the label in one of the phases as an indication of said polyvalent antigen, wherein said first receptor either comprises an oligomer of at least two antibody molecules or consists of a oligomer of antibody fragments selected from the group consisting of Fab or (Fab'"'"')2 fragments bound together which antibody molecules or antibody fragments all have specificity for the antigen to be determined.
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Abstract
The present invention provides a process for the quantitative determination of a polyvalent antigen by incubation with three different receptors of which the first (R1) and the third (R3) are present in liquid phase and are bindable with the antigen, the second receptor (R2) is present in solid phase and is bindable with receptor (R1), and receptor (R3) carries a label and does not cross-react with (R1) and (R2), separation of the solid phase from the liquid phase and measurement of the label in one of the phases, wherein a first receptor (R1) is used which consists of at least two antibody molecules or antibody molecule fragments bound with one another, at least one of which binds specifically with the antigen to be determined.
The present invention also provides a reagent for the quantitative determination of a polyvalent antigen containing three receptors (R1, R2 and R3) of which (R1) and (R3) are soluble and bindable with the antigen, (R3) is labelled and does not cross-react with (R1) and (R2), wherein (R1) consists of at least two antibody molecules or antibody molecule fragments bound with one another, at least one of which binds specifically with the antigen to be determined.
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21 Claims
- 1. Process for the quantitative determination of a polyvalent antigen comprising incubating a liquid sample containing said antigen with three different receptors, wherein the first receptor (R1) and the third receptor (R3) are in liquid phase and bind with the polyvalent antigen, and the second receptor (R2) is in solid phase and binds with (R1), wherein (R3) carries a label and does not cross react with (R1) and (R2) to form a solid phase complex of (R2), (R1), antigen and (R3), separating the solid phase from the liquid phase and measuring the label in one of the phases as an indication of said polyvalent antigen, wherein said first receptor either comprises an oligomer of at least two antibody molecules or consists of a oligomer of antibody fragments selected from the group consisting of Fab or (Fab'"'"')2 fragments bound together which antibody molecules or antibody fragments all have specificity for the antigen to be determined.
- 9. Process for the preparation of a receptor (R1) either containing at least two antibodies or consists of a oligomer of antibody fragments selected from the group consisting of Fab and (Fab'"'"')2 fragments bound together wherein all of said antibodies or antibody fragments have specificity for an identical antigen comprising reacting from 1/4 to 1/2 of the total amount of each antibody or antibody fragment with a di- or polyvalent linker to provide derivatives of said antibodies or antibody fragments thereof and linking two of said derivatives together under conditions which favor reaction of said linkers to form said receptor (R1).
- 11. Reagent for the quantitative determination of a polyvalent antigen comprising three receptors (R1, R2 and R3) wherein receptor (R1) and receptor (R3) are soluble and bind to the antigen to be determined, wherein receptor (R3) is labelled and does not cross react with receptor (R1) either and receptor (R2), and receptor (R1) comprises at least two antibodies or consists of a oligomer of antibody fragments selected from the group consisting of Fab and (Fab'"'"')2 fragments bound together, wherein all of said antibodies or antibody fragments specifically bind with the antigen to be determined, and wherein receptor (R2) is in insoluble form and binds to (R1).
Specification