We want to lyophilize 2 different nanobodies (different target and come from different source) and evaluate its affinity (pre and post lyophilization). Prior lyophilization, we resuspend the nanobodies in PBS or ammonium bicarbonate. After lyophilization, one nanobody lost its function, and the other partially reduced its affinity and specificity (cross-reacted). I read that this could happen to some proteins because of the buffer in which is resuspended, but seems strange because nanobodies are suposed to be very stable. Any idea?
Dear Eduardo,
There is a possibility that pH shift due to freezing will damage your samples. When you freeze the pH of buffer can change, a lot! I have heard researchers say that phosphate buffer can be one of the worst. You need to check the pH shift of your buffer when freezing and check that your nanobodies are OK with this pH shift.
There is a recorded webinar on this subject at:
https://www.spscientific.com/Webinars/Archives/
Look for:
Phase Transformations During Freeze-Drying: Potential Implications on Drug Product Performance
Occured: 12/12/2017 10:00 AM - 2:00 PM ET Presenter(s): Raj Suryanarayanan, Ph.D
You will have to register on the site to view the webinar.
Kind regards,
Rob
Hi Eduardo
Were you able to find a solution to this issue? if so what was it?
Thanks
Peter
Hi Peter,
I couldn't find a solution. Other researchers observed the same problem for nanobodies. But, if you want to concentrate them, Amicon ultra centrifugal filters works fine. If you want to store them momentarily - at leat for my case - I kept them at room temperature for 5 days keeping their properties (I needed to take them with me in a 24 hours trip, they work fine after that).
Best,
Eduardo
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