Re: [Histonet] CD3 clean with RB monos


I agree that the epitomics website is great for learning about this technology and I further agree it could be of help as has been mentioned before.  I am a bit skeptical of claims of a 10-fold better affinity than mouse monoclonals as a blanket statement.

At a previous meeting a while back we asked about claims of higher affinity and couldn't  get a sufficient (to our mind) response.  To me that response would be in the form of a side to side comparison, using the same immunogen, and with a host mouse and rabbit side by side each producing monoclonals to same target to see which might be better.  I believe the concept of a superior rabbit monoclonal technology in terms of being able to break immunological tolerance in the immunized systems, especially for some difficult targets, is probably right on.
But simply breaking tolerance is not the same as producing higher affinity antibodies.

For the blanket statement that rabbit monoclonals have higher affinity, I'd like to see data comparing them to their (identical) target in a mouse host and see data such as from Biacore, x-ray crystallography and that differences in somatic hypermutation are indeed making these higher affinity.
That being said, I agree that the rabbit monoclonals have great use and even more potential.  I've used several (many) that are far superior to their counter-part mouse monoclonals.  However, I've used (and heard of) a few that weren't as good. 
So while I like rabbit monoclonals, use them, advocate for them and endorse the suggestion you try them, I'm not convinced that  as a rule they have 10x higher affinity than do mouse monoclonals.

Ray Koelling
Phenopath Laboratories
Seattle, WA

 -------------- Original message ----------------------
From: "Mildred Fail" 
> We have had quite a problem with CD3s on bone marrow biopsies  being
> "messy" both with mouse monoclonals and rabbit polyclonals. Protein
> block is used. Diluting the Ab out further lost some cells in the lymph
> node control. We tried a  rabbit monoclonal. The staining is very
> specific and intense. The slide is beautifully free of extraneous
> staining. The higher dilution has not appeared to have effected the
> number of cells stained. Question       is why would the rabbit
> monoclonal produce a cleaner slide?
> Rena Fail  
> Rena Fail
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