Analysis of complement activation products in biological fluids using polyacrylamide gel electrophoresis

A biotin-labelled proteoglycan fraction which contained both GlcNAc and GalNAc was treated by sequential addition of chondroitinase AC/ABC and heparinase/heparitinase (Seikagaku, Tokyo, Japan) (Kato et al., 1985; Oike et al., 1080). A sharp band at the entrance of the separation gel in the untreated sample (Fig. 1; lane 1 ) is nearly abolished after chondroitinase treatment (Fig. 1: lane 2). A band in the molecular mass range of 150 kDa is unaffected. This band disappears after additional heparinase/heparitinase treatment and a sharp band in the range of < 2 9 kDa appears (Fig. 1: lane 3, see arrowhead). Thus, in accordance with the amino sugar analysis, the preparation contains both chondroitin sulphate and heraran sulphate. A fraction rich in GalNAc (Fig. 1; lane 4 ) was digested by chondroitinase. A smear in the range of 180-30 kDa disappeared leaving two well-defined bands of 89 kDa and < 29 kDa. Further treatment by heparinase/heparitinase and keratanase (Saikagaku) did not change the staining pattern (not shown). Thus, this preparation contained chondroitin sulphate only.