Figure 3

Fractionation of Heparin Byproducts via HIP peptide-1 Chromatography and Determination of ATIII-dependent anti FXa Activity. (A) A subsaturating amount of Hp byproducts in PBS was applied to 3 mls of a HIP peptide-1 affinity matrix, pre-equilibrated with PBS. Unbound materials were washed from the column by 15 ml of PBS and bound materials were eluted with a 0.15 to 0.70 M NaCl gradient. 1-ml fractions were collected and the amount of material and salt content in each fraction was quantitated as in "Materials and Methods." The X-axis corresponds to the fraction number while the left Y-axis denotes the total amount of GAG material in each fraction. The right Y-axis gives the NaCl concentration of each fraction. (B) Fractions 6, 19, 22, and 24 were tested in an ATIII-dependent anti FXa chromogenic assay as described in "Materials and Methods."