Pierce Chemical Company

Keld Sorensen

A Krishna Mallia

A rapid radioactive method of measuring enzymatic activity of a protein kinase is disclosed. The method is an improvement to existing methodology which involves phosphorylating a peptide substrate using .sup.32 P-ATP, adsorbing the phosphorylated peptide to a solid phase, washing the phase to remove non-adsorbed .sup.32 P-ATP, and measuring the radioactivity of the phosphorylated peptide adsorbed to the phase. The disclosed improvement uses a membrane as the solid phase and positions the membrane within a chamber to separate the chamber into a first and second region. Washing is accomplished with centrifugal force; the washed solution being forced through the membrane from the first region into the second region.

Rapid assay for radioactive determination of protein kinase activity

A process is described for isolating an enzyme-antibody conjugate, wherein the enzyme is horseradish peroxidase or alkaline phosphatase, from an aqueous mixture of said conjugate and unconjugated enzyme. The process involves contacting the mixture with a water insoluble stationary phase having the Ni.sup.+2 ion chelated thereto and binding said conjugate to the stationary phase. The phase containing bound conjugate is then washed to remove unbound enzyme. Thereafter the conjugate is eluted from the stationary phase and recovered in a form substantially free of the unconjugated enzyme.

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Roscoe, IL, United States of America

Keld Sorensen