Validate SDS-Page (Collagen Typ I)
Posted 02 November 2009 - 02:04 AM
Iīm new at this forum and have a little problem with a validation.
I used the search-function and couldnīt find any thread with this topic.
I have to validate the sds-page at work.
We are characterizing collagen typ I most of the time.
I never did a validation before so I read a lot and learned what to do.
So I get the information to vaildate a few important things.
Robustness is one of the easy to describe parameters.
I have big trouble in phrasing "Specificity" and a little trouble with "limit of detection".
About limit of detection: Made a series of measurements with descending percentage of collagen. So I can see at which level I can detect the last visible bands in the gel. Ok. But I have hassle to validate "Specificity". SDS-PAGE isnīt specific to collagen typ I, how to show that?
My question: Has someone managed a validation for characterizing collagen typI with SDS-Page recently and is willing to help me with this?
Maybe this is a dumb question for some of you pros....
Thank you for reading and sorry for my english.
Posted 02 November 2009 - 02:14 AM
Specificity covers within itself.. matrix interference (buffers run) and separation selectivity (in this you can show it can separate between dimer and monomer or alternatively any other HCP's give band at different position)
Also depends if you wanna use this test as just an identity test or a purity test!!!
any other opinions are welcome..
Posted 02 November 2009 - 02:33 AM
Good clear answer.
That helped a lot.
Posted 02 November 2009 - 02:58 AM
ya a great forum indeed!!!