[Histonet] RE: IHC antibody optimizing & validating

[Terri Braud]

CAP is very clear that in order to validate a new antibody, that: "once
the stain has been optimized, that for a well characterized antibody
with a limited spectrum of antigenic targets, like Chromogranin or PSA,
the validation can be limited.  A panel of 10 positive and 10 negative
neoplasms would be sufficient in this setting.  For an antibody that is
not well characterized and/or has a wide range of reported reactivity, a
more extensive validation is necessary.  The number of tissues tested
should in the circumstance be large enough to determine whether the
staining profile matches that previously described.  An exception to the
above requirements is that studies nay not be feasible for antigens such
as ALK that are only seen in rare tumors."
Thus sayeth CAP.
And if you're like me, I am not digging through all my cases to try to
come up with 30-40 neoplasms for each antibody, so I just order Tissue
Micro Arrays with the neoplasms I need.  20 positive and 20 negative
neoplastic tissues on one slide for easy staining and validation.  The
money you save on reagents to stain one little slide, more than makes up
for the cost of the slide.
I hope this helps.

Terri L. Braud, HT(ASCP)
Anatomic Pathology Supervisor
Holy Redeemer Hospital Laboratory
1648 Huntingdon Pike
Meadowbrook, PA 19046
Ph: 215-938-3676
Fax: 215-938-3874

Today's Topics:
   8. IHC antibody optimizing & validating (Davis, Cassie)
   
Message: 8
Date: Tue, 25 Mar 2014 09:51:07 -0400
From: "Davis, Cassie" <CDavis <@t> che-east.org>
Subject: [Histonet] IHC antibody optimizing & validating

Will you help me? I understand we are to use the known positives
controls that the manufactures' recommends in the package insert when
optimizing the stains, but I need to know what is your general procedure
for optimizing (how many different staining protocols do you test) and
validating a new antibody (how many different or "known" positive and
negative tissues do you test [predictive markers I understand are 20])?

Cassandra Davis
CDavis <@t> che-east.org
302-575-8095

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