Immobilisation of urease enzyme on PMMA (acrylic polymer)

Andy Greenhalgh andy.greenhalgh at localhost.localdomain
Sun Feb 20 07:09:16 EST 2000


I have developed a fibre optic fluorimeter which uses cheap acrylic
fibre rather than the more expensive silica variety. One of the assays I
am
working on utilises jack bean urease to metabolise urea. Ammonia is
liberated increasing the pH dependant fluorescence of a fluorophore in
the surrounding solution.
      
This approach works well, however the next step will be to immobilise
the urease onto the stripped fibre core. The instrumentation I have
designed
employs a phenomenon known as the evanescent wave to observe
fluorescence in the area immediately surrounding the fibre (to a depth
of several hundred nm) and can therefore monitor pH local to the enzyme.
      
My question is this: has anyone any suggestions/advice for methods of
covalently attaching enzymes onto to PMMA (acrylic polymer).
          
I am not familiar with the pros and cons of the various techniques for
functionalising PMMA and would be grateful for any advice that anyone
might have to offer.
      
NB I have avoided non-covalent attachment as the pH changes involved
(6.0-9.5) might result in some enzyme detachment.
      
Thanks




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