Protein Science, Vol 1, Issue 7 935-944, Copyright © 1992 by Cold Spring Harbor Laboratory Press

ARTICLE

Analysis of peptide mixtures by capillary high performance liquid chromatography: A practical guide to small-scale separations

M. T. DAVIS and T. D. LEE
Division of Immunology, Beckman Research Institute of the City of Hope, Duarte, California 91010

Capillary HPLC is a very effective means of separating small amounts of peptides and proteins. Capillary columns ranging from 0.01 mm to 0.5 mm in diameter can be constructed using recycled supports and inexpensive fused silica capillary tubing. Commercial pumping systems and UV detectors can be readily converted for operation in the flow rate range of 0.5-50 {mu}L/min. Detailed procedures are given for the construction of columns and UV detector flow cells. A mixture of peptides derived from the endo Lys C digest of horse heart cytochrome c was used to illustrate various aspects of capillary chromatography of peptides and compare the performance of various-sized capillary columns and UV detector flow cell types.
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