Vitamin B12 assay
AssigneeTritech Partners
InventorsMohammad T. Pourfarzaneh, George W. Katsilometes
Life of this patent
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Abstract
A competitive immunoassay for vitamin B.sub.12 using labeled monoclonal antibodies to the vitamin B.sub.12 binding site on intrinsic factor and labeled intrinsic factor.
Claims9
An immunoassay kit for determining the concentration of vitamin B.sub.12 in a sample comprising,
- a. monoclonal antibodies that are specific for the vitamin B.sub.12 binding site on intrinsic factor and block the binding of vitamin B.sub.12 to intrinsic factor said monoclonal antibodies being labelled with a first label;
- b. intrinsic factor labelled with a second different label;
- and
- c. a solid phase containing a capture ligand for one of said labels with the other label being a signal generating label for use in detection.
A complex wherein intrinsic factor having a first label is bound to a monoclonal antibody specific for the vitamin B.sub.12 binding site on intrinsic factor thereby blocking the binding of vitamin B.sub.12 to intrinsic factor, said monoclonal antibody having a second different label, where one of said labels is capable of being captured by a capture ligand on a solid phase and the other label is a signal generating label for use in detection.
A process for the determination of the concentration of vitamin B.sub.12 in a fluid containing vitamin B.sub.12 bound to binding proteins comprising the steps of:
- (a) treating a sample of the fluid to free vitamin B.sub.12 from the binding proteins;
- (b) contacting the sample with labelled monoclonal antibodies specific to the vitamin B.sub.12 binding site on intrinsic factor thereby blocking the binding of vitamin B.sub.12 to intrinsic factor and labelled intrinsic factor which is labelled with a different label from that used with the monoclonal antibodies, one of the labels being capable of being bound by a capture ligand of a solid phase and the other label being a signal generating label to form intrinsic factor:antibody and intrinsic factor:vitamin B.sub.12 complexes;
- (c) adding the solid phase containing the capture ligand for one of said labels to said complexes to form bound and unbound complexes;
- (d) separating the bound and unbound complexes and washing;
- and
- (e) measuring either the amount of bound complex or unbound complex to quantify the amount of vitamin B.sub.12 present in the sample.
Dependencies are inferred from the claim text.