Quantitation of collagen types I, III and V in tissue slices by capillary electrophoresis after cyanogen bromide solubilization

Capillary electrophoretic separation of proteins and peptides by ion-pairing with heptanesulfonic acid

“…Application of this separation principle enables a better resolution of the shorter peptides than in the case of the use of phosphate buffer only. In comparison with previously published results [7] we obtained better and faster separation of the shorter peptides than in the case when two-times longer capillary with phosphate buffer only was used. A better view of the CNBr peptide profile can be seen in Fig.…”

“…Separation of this complex mixture has to be made on a long capillary (ca. 60/70 cm long, see [7,9]). To demonstrate the applicability of the ion-pairing principle in the case of complicated peptide system we used in the current experiments two-times shorter capillary (30/37 cm).…”

“…The main peaks in the front of the slope are ␣ 1 (I)CB 2 , ␣ 1 (I)CB 4 , ␣ 1 (III)CB 2 , ␣ 1 (I)CB 5 , ␣ 2 (I)CB 2 and ␣ 1 (I)CB 3 . Detailed way of identification was published in our previous papers [7,9].…”

“…This sample contained fragments of both type I and III collagen and was prepared by the procedure described in our previous communication [7]. Briefly, samples (rat tail tendons) were incubated in 0.2 mol/l ammonium bicarbonate, pH 7.0, containing 25% (v/v) ␤-mercaptoethanol to reduce oxidised methionyl residues and after lyophilisation the samples were cleaved by CNBr in 70% (v/v) formic acid under nitrogen.…”

Capillary electromigration methods for the study of collagen

“…This method is also suitable for the determination of higher molecular mass peptides arising from the posttranslational modification of proteins (cross-links due to glycation reactions). The same relationships were demonstrated on the separation of CNBr peptides including low molecular mass peptides from collagens type III (␣ 1 (III)CB 2 ) and V (␣ 1 (V)CB 1 ) [23]. The separation of a mixture of these collagen fragments (including spiked markers of collagen types III and V) is demonstrated in Fig.…”