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enzyme kinetics in ELISA

Robert Burns burns at sasa.gov.uk
Tue Aug 29 04:11:52 EST 1995

Message from Isla Browning via Robert Burns( my E mail is out of order)

My son is writing up a school project in which he carried out 
experiments to calculate the Km of alkaline phosphatase acting on 
p-nitrophenylphosphate in both ELISA, and when the enzyme is free in 
solution. The experiments were carried out at 25C and at a pH of 9.8. 
The result was Km= 0.246 for the immobilised enzyme and 0.505 for the 
free solution. I would be grateful for any comments on these results. 
It means that there was greater affinity of the enzyme for the 
substrate when it was immobilised, which is not what we expected.

Isla Browning
Diagnostics and Molecular Biology Section
Tel 0131 244 8916

Robert Burns

Monoclonal Antibody Unit
Scottish Agricultural Science Agency
East Craigs

burns at sasa.gov.uk

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