Table 4.

Purification of VEGF165 from 6HFh8-VEGF165 fusion protein expressed in E. coli

Purification step	Concentration (mg/ml)	Volume (ml)	Total protein (mg)	Fusion protein (tag-free protein) (mg)	Purity (%)a	Yield (%)b
Crude extract	1.5 ± 0.0	100	147.5 ± 0.7	40.8 ± 0.2 (18.8 ± 0.1)	27.1	100
HisTrap, 5 ml	0.3 ± 0.0	88	30.4 ± 0.6	29.0 ± 0.6 (13.4 ± 0.3)	95.9	79.8
HiTrap SP, 5 ml	0.11 ± 0.0	11	1.3 ± 0.0	NA (2.9 ± 0.1)	80.4	6.1
HisTrap, 5 ml	0.04 ± 0.0	21	0.8 ± 0.0	NA (2.8 ± 0.1)	95.8	4.6
HiTrap SP, 5 ml	0.03 ± 0.0	6	0.2 ± 0.0	NA (0.2 ± 0.0)	> 99	0.9

The data are shown as mean ± standard deviation from triplicate independent experiments

^aPurity was analyzed by densitometry using ImageJ and C18 RP HPLC

^bYield was calculated by dividing the tag-free protein of each purification product by tag-free protein in crude extract