Separation of peptides by reverse-phase high-performance liquid chromatography using propyl- and cyanopropylsilyl supports |
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Authors: | P Tempst M W Hunkapiller L E Hood |
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Institution: | Division of Biology, California Institute of Technology, Pasadena, California 91125 USA |
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Abstract: | The separation of peptides and proteins by reverse-phase high-performance liquid chromatography with cyanopropylsilyl and large-pore propylsilyl supports, together with aqueous trifluoroacetic acid/acetonitrile gradients, was studied. Operating parameters (trifluoroacetic acid concentration, flow rate, and gradient slope) were evaluated using different enzymatic digests of horse cytochrome c and bovine serum albumin. Peptides ranging in size from five amino acids to 68 kDa could be separated on the propylsilyl column in a single chromatographic run. The cyanopropylsilyl column is suitable as a supplement to the use of the large-pore column for medium size (5-20 amino acids) peptides. The chromatographic supports and conditions presented here offer a simple, sensitive, and rapid separation system for a wide size range of peptides and proteins. They extend the versatility of separation methodology for these molecules. |
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Keywords: | protein purification peptide purification HPLC propylsilyl columns peptide sequencing trifluoroacetic acid |
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