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Position: Discussion forums > Bioscience biotechniques discussion forums > Proteomics and protein biochemistry forum
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PROBLEM WITH RP-HPLC PURIFICATION OF PEPTIDE
Posted by: SIVA (IP Hidden, New member, 0)
Date: July 2, 2002 02:50PM
I HAVE A PROBLEM WITH THE PURIFICATION OF 26 AMINOACID PEPTIDE PREPARED BY SOLID PHASE PEPTIDE SYNTHESIS. THE SEQUENCE IS NH2-CGGGEGGGEGGGEGGGEGGGEGGGEG-CONH2 THEY ARE PREPARED BY FRAGMENT CONDENSATION OF TETRA PEPTIDES. PEPTIDE IS NOT VISIBLE AS A PEAK IN TFA(PH2) AND AMMONIUM ACETATE (PH 5.5) BUFFERS. IN AMMOUNIUM BICARBONATE BUFFER (PH 7) IN C18 9.4*250mm COLUMN IT HAS RETENTION TIME OF ABOUT 3.5MINUTES AND IT COELUTES WITH OTHER DELETION PEPTIDES OF MY SOLID PHASE SYNTHESIS. ANY SUGGESTIONS ON DIFFERENT BUFFER OR COLUMN CONDITIONS??? THANKS, SIVAKUMAR
Re: PROBLEM WITH RP-HPLC PURIFICATION OF PEPTIDE
Posted by: Venkat r (IP Hidden, Unregistered user, )
Date: January 25, 2005 04:52PM
dear siva
the siva , have you checked your mp net pH, because the net pH of your mp may be causing the problem what you are facing, hence, i suggest you bring your mp pH as near as to 7
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