Characterization of bovine collagens using capillary electrophoresis - an alternative to slab gel electrophoresis

A capillary electrophoresis method was developed and characterized for anal yzing the spectrum of collagen subspecies in collagen preparations. The Bio -Rad CE-SDS protein kit was used for the dynamic sieving separation of coll agen subspecies in this CE method (DSCE). The optimized method utilized a 3 6 cm (or 24 cm) x 50 mu m uncoated capillary, electrophoretic injection at 10 kV for 10 s, a run voltage of 15 kV, a capillary temperature of 20 degre es C, and UV detection at 220 nm. A preliminary validation of the method wa s performed. The assay had good repeatability (RSDs for peaks were 1-5%), a nd responses were linear for assay solutions with collagen concentrations f rom 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 t o that generated by scanning of Coomassie-stained SDS-PAGE gels. (C) 2000 E lsevier Science B.V. All rights reserved.