A recombinant polymeric hemoglobin with conformational, functional, and physiological characteristics of an in vivo O2 transporter

被引:32
作者
Bobofchak, KM
Mito, T
Texel, SJ
Bellelli, A
Nemoto, M
Traystman, RJ
Koehler, RC
Brinigar, WS
Fronticelli, C
机构
[1] Johns Hopkins Univ, Sch Med, Dept Anesthesiol & Crit Care Med, Baltimore, MD 21287 USA
[2] Univ Roma La Sapienza, Inst Mol Biol & Pathol, Consiglio Nazl Ric, Rome, Italy
[3] Univ Roma La Sapienza, Dept Biochem Sci, Rome, Italy
[4] Temple Univ, Dept Chem, Philadelphia, PA 19122 USA
来源
AMERICAN JOURNAL OF PHYSIOLOGY-HEART AND CIRCULATORY PHYSIOLOGY | 2003年 / 285卷 / 02期
关键词
hemoglobin recombinant; hemoglobin synthetic; hemoglobin; polymeric; blood substitutes; transfusional fluids; artificial oxygen carriers; hemoglobin retention time; exchange transfusion; extravasation; focal cerebral ischemia;
D O I
10.1152/ajpheart.00037.2003
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
With the objective of developing a recombinant oxygen carrier suitable for therapeutic applications, we have employed an Escherichia coli expression system to synthesize in high-yield hemoglobin (Hb) Minotaur, containing alpha-human and beta-bovine chains. Polymerization of Hb Minotaur through S-S intermolecular cross-linking was obtained by introducing a Cys at position beta9 and substituting the naturally occurring Cys. This homogeneous polymer, Hb Polytaur, has a molecular mass of similar to500 kDa and was resistant toward reducing agents present in blood. In mice, the circulating half-time (3 h) was fivefold greater than adult human Hb (HbA). The half-time of autooxidation measured in blood (46 h) exceeded the circulating retention time. Hypervolemic exchange transfusion resulted in increased arterial blood pressure similar to that with albumin. The increase in pressure was less than that obtained by transfusion of cross-linked tetrameric Hb known to undergo renovascular extravasation. The nitric oxide reactivity of Hb Polytaur was similar to HbA, suggesting that the diminished pressor response to Hb Polytaur was probably related to diminished extravasation. Transfusion of 3% Hb Polytaur during focal cerebral ischemia reduced infarct volume by 22%. Therefore, site-specific Cys insertion on the Hb surface results in uniform size polymers that do not produce the large pressor response seen with tetrameric Hb. Polymerization maintains physiologically relevant oxygen and heme affinity, stability toward denaturation and oxidation, and effective oxygen delivery as indicated by reduced cerebral ischemic damage.
引用
收藏
页码:H549 / H561
页数:13
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