Characterization of bovine collagens using capillary electrophoresis.
A capillary electrophoresis method was developed and characterized for analyzing the spectrum of collagen subspecies in collagen preparations. The Bio-Rad CE-SDS protein kit was used for the dynamic sieving separation of collagen subspecies in this CE method (DSCE). The optimized method utilized a 36 cm (or 24 cm) x 50 microm uncoated capillary, electrophoretic injection at 10 kV for 10 s, a run voltage of 15 kV, a capillary temperature of 20 degrees C, and UV detection at 220 nm. A preliminary validation of the method was performed. The assay had good repeatability (RSDs for peaks were 15%), and responses were linear for assay solutions with collagen concentrations from 0.125 to 1.25 mg/ml. The DSCE electropherogram of bovine skin collagen provided a profile of subspecies similar in number and relative abundance to that generated by scanning of Coomassie-stained SDS-PAGE gels.
Chang
P
P
Martrix Pharmaceutical, Fremont, CA 94555, USA.
Kuan
S
S
Eberlein
G
G
Burke
D
D
Jones
R
R
D000818:Animals; D002417:Cattle; D003094:Collagen / Q000032:analysis*; D019075:Electrophoresis, Capillary; D004591:Electrophoresis, Polyacrylamide Gel; D012867:Skin / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet
D003094:Collagen