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Separation of peptides by reverse-phase high-performance liquid chromatography using propyl- and cyanopropylsilyl supports

1984; Elsevier BV; Volume: 137; Issue: 1 Linguagem: Inglês

10.1016/0003-2697(84)90369-5

1096-0309

Paul Tempst, Michael W. Hunkapiller, Leroy Hood,

thermodynamics and calorimetric analyses

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.