Reverse Phase HPLC of protein in neutral solvent system

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antibiotics
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Reverse Phase HPLC of protein in neutral solvent system

Hi,

    Glad to find this forum with so many useful information.
   I am working a protein adduct characterizing project. I was able to seperate the target protein by a RP HPLC (C4 column, Solvent A: water with 0.1% TFA; B Acetonitrile with 0.1 TFA), gradient 30%B to 95%B.
   However, the adduct seems not stable under acidic condition. I am thinking if it is possible to use some netural solvent system to isolate my protein? Thanks.

qiang wang
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you can neutralize the eluted fractions with 1M Tris-HCl, pH 8.5 to desired pH immediately.

g a
g a's picture
Have you tried using milder

Have you tried using milder ion pairing agents like HCOOH ? It works better many times than commonly used TFA.