Radioassay kit for and method of determining methotrexate

Details Radioassay kit for and method of determining methotrexate Radioassay kit for and method of determining methotrexate

G - Physics

01

N

150/9

G01N 33/48 (2006.01) C12Q 1/32 (2006.01) G01N 23/00 (2006.01) G01N 33/94 (2006.01)

Patent

CA 1075135

ABSTRACT OF THE DISCLOSURE A radioassay system for the determination of methotrex- ate in biological fluids based on the competitive binding of labeled and unlabeled methotrexate to the enzyme dihydrofolate reductase. Samples of unknown methotrexate level are mixed with a labeled methotrexate particularly an I125 labeled methotrexate. A portion of the total methotrexate present is bound by the addi- tion of enzyme, and the unbound methotrexate is removed with char- coal. The level of bound I125 labeled methotrexate is measured in a gamma ray counter. To calculate the methotrexate level of the unknown samples, the displacement of bound labeled methotrex- ate caused by the unknowns is compared to the displacement caused by known methotrexate standards.

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