Extracting secreted peptides from nutrient broth. -Reply

Henry Wolfe (hnwo@americas.nycomed.com)
Fri, 09 Oct 1998 11:06:38 -0600

Treat the supernatant in a manner to precipitate macormolecules and
leave the peptide in solution, then run the first of several chromatogrphic
steps.

In my suggestions, I assume you have a bioassay or immunoassay for
the peptide.

Two quick thoughts:
1.) Pellet proteins, etc with 50% aq. ethanol, rotovap the spnt to remove
the ethanol and purify by IEX (if possible) or SEC, followed by RP-HPLC.

or

2.) Pellet proteins, etc with appropriate NH4SO4 concentration to leave
peptide in solution. Run peptide fraction through HIC, followed by
RP-HPLC

good luck,

Hank Wolfe
Nycomed Amersham
Philadelphia, Pa.