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- W2894434846 abstract "Peptide purification via high-performance liquid chromatography (HPLC) and solid-phase sequencing were integrated to form a system allowing the determination of complete sequence information on a microscale without the use of radiolabels or modified phenylisothiocyanate. Mixtures of peptides (500 pmol to 10 nmol) resulting from proteolytic digestion or chemical cleavage were applied directly to reverse-phase columns. The columns, equilibrated in either 10 m m K P i or 0.05% trifluoroacetic acid, were then developed using acetonitrile gradients. Eluates were monitored nondestructively by direct ultraviolet detection at both 214 and 254 nm. Each peak was collected as a discrete fraction, and purity was assessed by amino acid analysis prior to covalent attachment to a solid support for sequence analysis. Activation of the peptide carboxyl terminus via a water soluble carbonyldiimide was the solid-phase coupling method used 90% of the time. Coupling yields averaged 52% of starting material. Sequence analysis was performed in the range 100 pmol to 4 nmol of coupled peptide. Phenylthiohydantoin-amino acids were identified by reverse-phase HPLC using ultraviolet detection." @default.
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- W2894434846 date "1983-01-01" @default.
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- W2894434846 title "Application of High-Performance Liquid Chromatographic Peptide Purification to Protein Microsequencing by Solid-Phase Edman Degradation11This paper was presented at the International Symposium on HPLC of Proteins and Peptides, November 16–17, 1981, Washington, D. C." @default.
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- W2894434846 doi "https://doi.org/10.1016/b978-0-12-335780-9.50028-0" @default.
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