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Disuccinimidyl suberate cross-linked hemoglobin as a novel red blood cell substitute

Science china(Life Sciences) 2005年第1期48卷 49-60页

作者： lu xiuling, zheng chunyang,{2.,{3., chinese academy of Sciences, Beijing 100080, china School of Life Science and Technology, Beijing University of Chemical Technology, Beijing 10002., china national key {2. of {3. engineering Institute of Process Engineering Chinese Academy of Sciences Beijing100080 China School of Life Science and Technology Beijing University of Chemical Technology Beijing100029 China

Disuccinimidyl suberate (DSS) intramolecularly cross-linked hemoglobin (Hb) was developed as a novel red blood cell substitute. A multi-angle laser light scattering detector coupled with size exc{3.sion HPLC was applied to determine the molecular weight of the modified Hb. SDS-PAGE was also used as a complement. It was proved that 83.8% of the product was intramolecularly cross-linked Hb with weight-average molecular weights (Mw) of 67.5 kD, 12. was dimeric Hb with Mw of 146.6 kD, and 4.2. was trimeric Hb with Mw of 3.6.4 kD. The tetramer structure of the cross-linked Hb was stable as shown in size-exc{3.sion chromatography using a mobile phase containing 1 mol/L MgCI2. Analysis by LC-MS demonstrated that the reaction of DSS with Hb mainly took place between the two a subunits within a Hb molecule, resulting in stabilization of the tetramer structure. However, the cross-linking was not site-specific. The P50 of the cross-linked Hb decreased from 2..8 mmHg to 14.3.mmHg, and the Hill coefficient decreased from 2.2. to 1.41. Result of isoelectric focusing showed that the pi of DSS cross-linked Hb was in the range of 4.6-5.2. similar to that of serum albumin. The safety of DSS cross-linked Hb was favored by animal tests on rats and guinea pigs. Exchange transfusion experiment with DSS cross-linked Hb using rats as a model indicated no pressor effect or other significant side effects. The characteristics and properties of DSS cross-linked Hb were also compared with that of diaspirin cross-linked Hb reported in the literature.

关键词： hemoglobin, disuccinimidyl suberate, cross-link, red blood cell substitute.

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