Chromatography Forum

Dear collegues,

I have not much experience of LC-analysis of proteins, but I would like to test ion-exchange for analysis of FSH-hormone. We are currently using SEC or reversed-phase - but I am curious of how it looks using an orthogonal separation technique.

Is there a the state-of-the-art column for this kind of separation? Being at a preferable pH of pH 7.4, I assume that anion-exchange is the best option (?). Any suggestions of a good starting point is appreciated.

Purification of FSH seems to be done regularly on DEAE resins (weak anion exchangers).

This might be a starting point for you:
joe.endocrinology-journals.org/cgi/reprint/165/2/185.pdf

Thanks Tom,

The article and references are quite old, and they concerns mostly larger volume of sample (i.e purification).

I am more looking for a state-of-the-art anion-exchange analytical column to put on my HPLC. I have had some good experiences of the Superdex SEC columns (Amersham), and they have a range of ion-exchange columns that I might try.

They have a column called Mini Q 4.6/50 PE. I would be glad to hear any experiences of this column!

(The main objektive of my test would be to separate FSH from the different oxidation products of FSH, but also to see if any unknowns appear)

Mattias,

I've had quite succesful protein separations on a Dionex ProPac SAX-10 (250 x 4 mm)
If you drop me a mail, I can suggest some starting conditions etc. to you.

dancho.dikov@stella-chroma.com

Best Regards