[Histonet] RE: IHC antibody optimizing & validating

[joelle weaver]

TMAs are great if that is one of your options, but unless you make them you have the PA issues that might enter in. CAP does gives specifics, but  I was just giving the information and process that is currently in place where I presently work to hopefully help, not trying to supersede CAP guidelines for sure- where would that land me?




Joelle Weaver MAOM, HTL (ASCP) QIHC
 
> Date: Tue, 25 Mar 2014 15:34:30 -0400
> From: tbraud <@t> holyredeemer.com
> To: histonet <@t> lists.utsouthwestern.edu
> Subject: [Histonet] RE: IHC antibody optimizing & validating 
> 
> 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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