Forum :: Removing ligand


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These are the posts of the old forum. It was not possible to transfer the user data, so they are missing in most of the posts. For new questions, go to the general discussions.
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TOPIC: Removing ligand

Removing ligand 15 Aug 2006 02:00 #1

Is it possible to remove the ligand from the CM5 chip?? I have a protein immobilised using amine coupling reagent and want to remove the protein so i can reuse the chip. If it is possible to remove the protein, will the regenerated chip be safe to use in the BIAcore machine? I do not want to cause any damage to the machine.

Amino coupling 15 Aug 2006 02:00 #2

Well, the amino coupling is a covalent immobilization method so, I am afraid you should break the covalent bond in order to eliminate the ligand from the surface. Then, it is considered an irreversible immobilization method and you have to use a new flow cell for a new experiment.

Amino coupling 15 Aug 2006 02:00 #3

Please look at the Download - Howto section en lookup de document about 'How to optimize regeneration'.
In here you will find a protocol to strip de your ligand from the sensorchip and to recodition the sensorchip surface.

regenerating chip 15 Aug 2006 02:00 #4

Thanks

I read the paper. To regenerate the chip you have to remove it from the casing and soak in Pronase solution to cleave protein from the dextran layer. A better way would be to inject Pronase onto the chip, however i cannot work out how to do this using the BIAcore software. Does anyone know how i could inject 30ul of Pronase solution into the BIAcore chip .??

regenerating chip 15 Aug 2006 02:00 #5

I think it is better to do the procedure outside the machine. Why incubating in the machine in stead of in a 50 ml tube. You have to clean your machine very thorougly to get rid of the pronase.

regenerating chip 15 Aug 2006 02:00 #6

Thanks for your reply. I am new to this. I was worried about damaging the chip if i removed it from the casing. Also I was worried that if i damaged the chip, the damaged chip could cause damage to the BIAcore optical detection system. It would be much easier to inject 60ul of Pronase onto the chip and leave it there overnight and then remove using NaOH. If NaOH was injected this would remove any Pronase.
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