[Histonet] RE: therapeutic antibody detection in mouse--Histonet Digest, Vol 60, Issue 33

From:"Nerissa Mendoza"


Isn't Herceptin a humanized mAb?  I believe you need to detect it with
an anti-human antibody (rather than mouse)?

Nerissa Mendoza
Senior Research Associate I
Preclinical Research & Development
2910 Seventh Street
Berkeley, CA 94710


Message: 3
Date: Wed, 19 Nov 2008 14:22:57 -0500
From: "Perry, Samuel" 
Subject: [Histonet] therapeutic antibody detection in mouse
Content-Type: text/plain; charset="us-ascii"

Hi All,
I am wondering if anyone has experience showing therapeutic monoclonal
antibody binding in mouse xenografts (human tumor cells injected
subcutaneously in SCID mice and then dose treated with therapeutic
antibody injected intraperitoneally).  For instance, can trastuzamab
(Herceptin) binding be seen in a breast tumor xenograft treated with
Herceptin?  I have tried using Dako's goat-anti mouse dextran HRP
conjugated secondary antibody developed with DAB.  I also tried using an
Abcam goat-anti mouse antibody conjugated with biotin, then adding
Vectors elite ABC, then developing with DAB.  Both times I used FFPE
tissue.  In both techniques, results showed minimal to staining and
showed no difference between mice treated with antibody and control mice
treated.  I would appreciate if anyone knew of published papers or have
experience using IHC to show therapeutic antibody bound to tumor cell in
mouse xenografts. 
Thanks -Sam 

Research Technician
Dana Farber Cancer Institute
Boston MA.

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