Purification stages Chromatographic Yield Specific fraction (mg) hemolytic activity (HU/mg) Extraction - 1700 212 Ion-exchange chromatography on Q1 543 572 Q-Sepharose Ion-exchange chromatography on SP1 178 1620 SP-Sepharose Hydrophobic interaction PS2 62 4510 chromatography on Phenyl-Sepharose Gel filtration on Superdex 75 S3 16 13.650 Purification stages Total hemolytic Recovery activity (HU) of HU (%) Extraction 360,400 Ion-exchange chromatography on Q-Sepharose 310,600 87 Ion-exchange chromatography on 288,400 80 SP-Sepharose Hydrophobic interaction chromatography on 279,600 76 Phenyl-Sepharose Gel filtration on Superdex 75 218,400 61 HU: hemolytic unit
. [FIGURE 1 OMITTED]
The MeOH and 1:1 DCM: MeOH, extracts on human AB blood group and cow blood showed maximum of 0.16 specific Hemolytic Unit (HU) and minimum of 0.01 specific Hemolytic Unit was recorded in goat blood from 1:1 DCM: MeOH extract (Fig.
Hemolytic activity of extracts the MeOH and 1:1 MeOH: DCM extracts on human AB blood group and cow blood showed maximum of 0.16 specific Hemolytic Unit (HU), followed by 0.08 HU observed in MeOH extracts on chicken, cow, human A & B erythrocytes.
The three hemolytic units had a mean (SD) HBDH concentration of 2585 (1797) U/L (1496-4660 U/L) with no significant difference after heating to 35[degrees]C [2160 (1107) U/L; 1270-3400 U/L].
In the three hemolytic units, 0.1 (0.04)% of the events were in R2 (minimum-maximum, 0.11-0.19%).