Chemically modified porcine hemoglobins and their biological properties.
Hemoglobin cross-linked with small molecular modifiers turns out to be more stable. Modifications of proteins with polyethylene glycol (PEG) have been proven to enlarge the molecular size of proteins, to prolong their retention time in the circulation as well as blunt immune reactions. In the present study, the optimal conditions for porcine hemoglobin (pHb) modification with bis (3, 5-dibromosalicyl) fumarate (DBBF) and PEG were evaluated. The derivative of DBBF cross-linked pHb (DBBF-pHb) showed improved oxygen affinity and the ability to resist the dissociation of the alpha2beta2 tetramer compared with the natural protein. DBBF-pHb was then bound to the activated PEG. The results indicated that the pHb modified with DBBF and PEG had more stable tetrameric conformation with a molecular weight of 107000. Their oxygen half-saturation pressure (P50) is around 3.33 kPa, which approximates the physiological P50 of human red blood cells. Both routine and reinforced immunizing methods were adopted to study the immunogenicity of modified products and the results showed that the products had very low immunogenicity evaluated by enzyme-linked immunoadsordent assay (ELISA). Somewhat beneficial effects were shown in the treatment of hemorrhagic shock where modified hemoglobin solutions were used as resuscitation fluids in the hemorrhagic shock Sprague-Dawley (SD) rats model.
College of Material and Chemical Engineering, Zhejiang University, Hangzhou 310027, China.
Cai
C
Jin
Meng
M
Wen-Fang
Hong
H
Min
Cen
C
Pei-Lin
Yuan
Y
Zhong-Yi
D000494:Allosteric Regulation; D000818:Animals; D001241:Aspirin / Q000031:analogs & derivatives* / Q000737:chemistry*; D001802:Blood Substitutes / Q000737:chemistry / Q000378:metabolism; D003432:Cross-Linking Reagents / Q000737:chemistry; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D006454:Hemoglobins / Q000737:chemistry* / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D008297:Male; D008970:Molecular Weight; D010100:Oxygen / Q000378:metabolism; D011092:Polyethylene Glycols / Q000737:chemistry*; D000091402:Prohibitins; D051381:Rats; D017207:Rats, Sprague-Dawley; D012771:Shock, Hemorrhagic / Q000473:pathology; D013552:Swine
D001802:Blood Substitutes; D003432:Cross-Linking Reagents; D006454:Hemoglobins; C000717328:PHB protein, human; C000717332:Phb protein, rat; D000091402:Prohibitins; C021630:bis(3,5-dibromosalicyl)fumarate; D011092:Polyethylene Glycols; D001241:Aspirin; D010100:Oxygen