Rapid fractionation of collagen chains and peptides by high-performance liquid chromatography.
A strategy was developed, using a Pharmacia Fast Protein Liquid chromatography (FPLC) system, for the rapid preparation of the alpha-chains, cyanogen bromide peptides and tryptic peptides of type I collagen obtained from tissues and cultured fibroblasts. Collagen alpha-chains were prepared using a C18 PEP-RPC reverse-phase column and volatile solvents. Preliminary Superose 6 gel permeation chromatography was used to separate the crosslinked beta- and gamma-chains from the alpha-chains of tissue collagen samples. A Mono S cation-exchange column was used to resolve all of the major type I collagen cyanogen bromide peptides including the alpha 1(I)CB7 and CB8 peptides, which have not been well resolved by previously published methods. Collagen tryptic peptides were chromatographed on the PEP-RPC reverse-phase column.
J F
JF
Bateman
T
T
Mascara
D
D
Chan
W G
WG
Cole
D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003094:Collagen / Q000302:isolation & purification*; D003488:Cyanogen Bromide; D005347:Fibroblasts / Q000032:analysis; D006801:Humans; D010446:Peptide Fragments / Q000302:isolation & purification*; D012867:Skin / Q000032:analysis; D014357:Trypsin
D010446:Peptide Fragments; D003094:Collagen; D014357:Trypsin; D003488:Cyanogen Bromide