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Let's NOT use gradient elution to get the high-affinity antibodies we need f

affinity chromatography gradient elution antibody management protocol

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3 replies to this topic

#1 Michael Starr

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Posted 17 March 2016 - 12:47 PM

My fucking supervisors decided NOT to do a common-sense gradient elution to enrich for high-binding strength antibodies in our affinity chromatography runs. And I'm too spineless to argue for it. I've never done it before myself, but I've seen plenty of examples where it yields multiple peaks corresponding to different binding affinity.

 

Fuck fuck fuck fuck fuck.

 

So dumb.



#2 mdfenko

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Posted 17 March 2016 - 01:52 PM

for a number of reasons, gradients may not be 100% reproducible.

 

You could run a linear gradient to find out approximate elution conditions then use a step gradient for subsequent elutions.


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#3 Ameya P

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Posted 18 March 2016 - 12:23 AM

Michael, 

 

You should consider speaking to your supervisors regarding their method of choice. They are senior in the field and there might be sufficient evidence with them for not doing the method (like mdfenko mentioned). 

 

Swearing here, won't do much. (Unless your supervisors are also on the forum :P

 

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#4 Michael Starr

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Posted 18 March 2016 - 08:12 AM

Thanks guys, for the advice and for quenching my ego. This is a venting forum, though, so swearing helps with that. I doubt my supervisors browse these forums; there aren't many members to begin with, and they're busy writing FDA reports and stuff.

 

Thanks for giving credit to their experience. Maybe they know better and I'm just not experienced enough. Good reality check.

 

Thanks.







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