Enzyme immobilization.

bford barry.ford at drdc-rddc.gc.ca
Mon May 16 15:19:42 EST 2005


In article <d69f9v$irg$1 at inews.gazeta.pl>, 
chris.b.nospam at nospam.interia.pl says...
> Hi,
> This may be stupid and/or simple question but I can't find answer...
> Can I and how I can bind protease (pepsin, trypsin) to chitosan or sephadex?
> I heard about binding enzyme to chitosan using glutaraldehyde and to
> sepharose using cyanogen bromide.
> Does anybody have good and cheap method to share with protein science
> freshman? :)
> 
> And one more thing - what is stability of such an enzyme? After how much
> time or how many use it looses activity (below "resonable" level)?

You can use glutaraldehyde to link anything to anything, and that's 
really the problem, low specificity.  Often times your target will stick 
to inself. Similar problems with CnBr. OTOH, you can monitor the 
reaction by just running sds gels of the rxn products. You'll see a 
shift in apparetn MW if the crosslinking works. CnBr activated resins 
are an excellent choice, if you can afford them. 

Given this caveat, glut is cheap (use only EM grade, not fixation 
grade), and I've found activity is well preserved. Stability will be  a 
function of storage, microbial contamination, and shit ass luck.
Probably no worse than your normal unlinked preps.

As to methods....lots of generic glut recipes out there. Have fun.
B.



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