Phage screening and regeneration - Forum


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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: Phage screening and regeneration

Phage screening and regeneration 25 Apr 2012 02:00 #1

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Anyone have experience with using SPR for phage screening...I am having some issues with regeneration and was wondering if anyone may have suggestions. I have tried low pH glycine 10mM, High pH glycine, 50-100mM HCl (which worked initially then stopped working). I am using an X100, use the regeneration function for 30seconds. I have avoided long dissociation times, but anyone have experience with longer regen times. When phage panning on plates, usually dissociation is about 10 minutes. Any help would be appreciated.
Thanks
rhaspel

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Phage screening and regeneration 25 Apr 2012 02:00 #2

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Hi,

I don't have experience with phage screening but have some general information about regeneration.

The regeneration pulse should be short. I have pulses of 10 - 20 seconds max. If this is not working you should look for another solution. Read: www.sprpages.nl/kinetics/regeneration.html and check the references for more idea's.

I'am not sure what you mean with long dissociation times. If your system is fully dissociated in 10 minutes, then don't bother with regeneration. For tight binders (low kd value) it can take a lot more time to dissociate fully. Then regeneration is an option.

Arnoud

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