[Preparation of bovine lipid-free hemoglobin].
A new method for preparation of Hb solution free of stromal lipid was described. Almost all the lipid in fresh hemolysate of bovine red blood was removed with hydrophobic interaction chromatography (HIC) in the presence of 2% PEG4000, 5% PEG4000, 2%PEG10000 or 5%PEG10000. With the adding of 5%PEG4000, the 80% of recovery of Hb in HIC was obtained and the maximum lipid absorbed by hydrophobic medium, butyl agarose -6B was 86.6 mg/mL. The activity (P50) of hemoglobin preparation was 3386.4 Pa torrs, and the Hill number was 2.54, which were near to that of the native red blood cells. The mechanism of removing lipid by HIC and the function of PEG in the process were discussed.
Zhao
Dong-Xu
DX
National Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, Beijing 100080, China. zhaodx1@yahoo.com
Wei
Xin-Gui
XG
Gu
Zhen-Yu
ZY
Zhang
Gui-Feng
GF
Su
Zhi-Guo
ZG
D000818:Animals; D002417:Cattle / Q000097:blood*; D006454:Hemoglobins / Q000302:isolation & purification*; D057927:Hydrophobic and Hydrophilic Interactions; D008055:Lipids / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000494:pharmacology
D006454:Hemoglobins; D008055:Lipids; D011092:Polyethylene Glycols