Heparinase III digestion in the presence or absence of MIP1α: isolation of the MPD.

³H-labeled HS chains were treated with heparinase III in the absence (A) or presence (B) of equimolar quantities of MIP1α (BB10010) as described in “Materials and methods.” Digests were analyzed by chromatography on a Biogel P10 column. The void (Vo) in panel B represents the MPD. (C) Molecular sizes of native HS chains (▪) and MPD (●) were compared by gel filtration on Sepharose Cl-6B. Vo indicates void volume; Vt, total volume. Distinct oligosaccharide peaks are labeled according to the number of monosaccharide units. (D) MPD (●) or intact HS (▪) were applied to an Affi-Gel MIP1α (BB10010) column in 0.15 M NaCl and eluted with a stepwise NaCl gradient (^{. . . . . . .}). Each graph is representative of 2 or more experiments.