[Cytometry] Anti human IgG2b secondary

Nebe-Von-Caron, G g.nebe-von-caron at alere.com
Wed Apr 10 04:33:00 EDT 2013


It is actually a good idea to test the binding of the second business end of your antibody in a different species. It was probably one of the key failures of the TGN1412 trial not to recognise that the different processing of the Fc end is a monkey business. They should really have noticed as they knew their in vitro experiments did not work if they used a Fab2

Guess you already picked up

http://www.jimmunol.org/content/early/2012/03/28/jimmunol.1200090.full.pdf

I would try directly labelling of the IgG2b with any of the commercially available kits. The advantage of that approach is that you can directly measure the binding affinity difference between human and monkey in situ using your flow cytometer even with your dedicated reagent. In addition you should look at their effect as immune-complexes and also make sure you remove them.

Have fun

Gerhard Nebe-von-Caron  


-----Original Message-----
From: cytometry-bounces at lists.purdue.edu [mailto:cytometry-bounces at lists.purdue.edu] On Behalf Of Szekeres, Karoly
Sent: 09 April 2013 18:34
To: cytometry at lists.purdue.edu
Subject: [Cytometry] Anti human IgG2b secondary

Dear all-
We are trying to identify binding of a human IgG2b antibody binding on Rhesus macaque monocytes.  I wonder if anyone could suggest a secondary that would recognize the human, but not the macaque antibodies.
Best,
Charlie

Charles Szekeres,
University of South Florida, Tampa
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