Method for measuring glycosyltransferase activity

Details Method for measuring glycosyltransferase activity Method for measuring glycosyltransferase activity

C - Chemistry, Metallurgy

12

Q

C12Q 1/48 (2006.01)

Patent

CA 2147887

A method of assaying for glycosyltransferase activity in a sample. In a first step, a sample is reacted with a first sugar don- or and an acceptor substrate to produce a transferase product. The first sugar donor and acceptor substrate are selected such that the sugar from the first sugar donor is capable of being transferred to the acceptor substrate in the presence of the glycosyl trans ferase to be assayed. In a second step, the transferase product is reacted with a second sugar don? having a sugar which is la- belled with a labelling agent and an enzyme which is capable of transferring the sugar from the second sugar donor to the trans- ferase product to produce a labelled transferase product and which has a higher affinity for the glcosysyltransferase product compared to the affinity of the glycosyltransferase for the acceptor substrate. The labelling agent activity of the labelled transfer ase product or unreacted second sugar donor is assayed to determine transferase activity in the sample. A kit for assaying for gly- cosyltransferase activity in a sample is also described.

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