Abstract The present invention provides a process for the determination of an immunologically-bindable substance by immunochemical methods in heterogeneous phase, wherein (a) a sample solution containing the substance to be determined is incubated with (b) a solid phase-bound antibody against the bindable substance to be determined (c) a conjugate of the bindable substance to be determined and a Fab fragment of an antibody directed against a determinable enzyme in a definite amount and (d) the enzyme which corresponds to the antibody contained in the conjugate (c), the phases are separated and the enzyme activity is measured in one of the two phases. The present invention also provides a reagent for the determination of an immunologically-bindable substance, wherein it contains a solid phase-bound antibody against the bindable substance to be determined, a conjugate of the bindable substance to be determined and a Fab fragment of an antibody directed against a determinable enzyme, a determinable enzyme which binds immunologically with the Fab fragment contained in the conjugate and buffer.
A process for determining an immunologically bindable substance comprising: (a) incubating a liquid sample solution containing the substance to be determined with a solid phase antibody against the bindable substance to be determined, which solid phase antibody is present in an amount insufficient to bind all of said bindable substance, a conjugate of the bindable substance to be determined and an Fab fragment of an antibody directed against a determinable enzyme, said conjugate being present in a predetermined amount and a sample of said determinable enzyme, (b) separating said solid and liquid phases, and (c) measuring the enzyme activity in one of the two phases as a measure of bindable substance in said liquid sample.
Claim 2
Dependent Claim
115 words
A process for determining an immunologically bindable substance comprising: (a) incubating a liquid sample solution containing the substance to be determined with a solid phase antibody against the bindable substance to be determined, which solid phase antibody is present in an amount insufficient to bind all of said bindable substance, a conjugate of the bindable substance to be determined and an Fab fragment of an antibody directed against a determinable enzyme, said conjugate being present in a predetermined amount and a sample of said determinable enzyme, (b) separating said solid and liquid phases, and (c) measuring the enzyme activity in one of the two phases as a measure of bindable substance in said liquid sample.
Claim 3
Dependent Claim
25 words
The process of claim 1, wherein a predetermined amount of the determinable enzyme is used and the enzyme activity is measured in the liquid phase.
Claim 4
Dependent Claim
25 words
The process of claim 1 wherein the antibody against the bindable substance is bound to the solid phase adsorptively, covalently by polymerising or by cross-linking.
Claim 5
Dependent Claim
13 words
The process of claim 1 wherein the determinable enzyme is β-galactosidase or peroxidase.
Claim 6
Dependent Claim
14 words
The process of claim 1 wherein the Fab fragment is a monoclonal Fab fragment.
Claim 7
Dependent Claim
26 words
The process of claim 1 wherein the determinable enzyme is β-galactosidase and the enzyme is determined colorimetrically using a measuring system comprising nitrophenylgalactoside, dinitrophenylgalactoside or aminophenylgalactoside.