A method for purification and viral inactivation of human placenta hemoglobin.
For pilot-scale manufacturing of hemoglobin-based oxygen carrying drugs, we should get highly pure and viral inactivated hemoglobin (Hb) at high recovery. In our method, placenta hemoglobin (PHb) solutions were purified by heating in the presence of reducing agent and deoxygenating conditions so that heat-sensitive proteins were selectively precipitated and virus was inactivated. The optimum preparative condition resulted in highly purified PHb solution (>99% pure) with approximate 90% recovery and less than 2% of MetHb content, maintained oxygen carrying capacity, residual phospholipids less than 1 ppm, free of endotoxin, bacteria, type A&B antigens and virus. Finally, we compared the efficacy of blood exchange on rat with poly-PHb and poly-Hb from adult blood. The results showed no significant difference between two products. Therefore, the placenta Hb obtained from this method could be supplied as materials for oxygen carrying drugs.
Key Laboratory of Drug Targeting of Education Ministry, West China School of Pharmacy, Sichuan University, Chengdu, China.
Tao
T
Li
Rong
R
Yu
Hong-Hui
HH
Zhang
Hong
H
Wang
Wei-Guang
WG
Liang
Xiao-Ming
XM
Yang
Cheng-Min
CM
Yang
D000818:Animals; D005079:Excipients / Q000494:pharmacology; D005260:Female; D006454:Hemoglobins / Q000187:drug effects / Q000302:isolation & purification*; D006358:Hot Temperature; D006801:Humans; D006863:Hydrogen-Ion Concentration; D010920:Placenta / Q000737:chemistry*; D000091402:Prohibitins; D051381:Rats; D019163:Reducing Agents / Q000494:pharmacology; D038301:Virus Inactivation
D005079:Excipients; D006454:Hemoglobins; C000717328:PHB protein, human; C000717332:Phb protein, rat; D000091402:Prohibitins; D019163:Reducing Agents