Poly(ethylene glycol)-conjugated human serum albumin including iron porphyrins: surface modification improves the O2-transporting ability.
Poly(ethylene glycol)-conjugated human serum albumin including iron porphyrins: surface modification improves the O2-transporting ability.
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DOI：
10.1021/bc050315
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发表时间：
2006-02
影响因子：
4.7
通讯作者：
Yubin Huang;T. Komatsu;Rong-Min Wang;A. Nakagawa;E. Tsuchida
中科院分区：
文献类型：
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作者：
Yubin Huang;T. Komatsu;Rong-Min Wang;A. Nakagawa;E. Tsuchida
由人血清白蛋白组成的人工载氧血红素蛋白，包括四(邻氨基苯基)卟啉铁(II)（Fe4P或Fe3P）[HSA-FeXP]已被马来酰亚胺或琥珀酰亚胺封端的聚乙二醇（PEG）修饰，并且所形成的PEG生物缀合物已被物理化学表征。 2-亚氨基四氢噻吩 (IMT) 与 Lys 的氨基反应产生活性硫醇基团，该硫醇基团与 α-马来酰亚胺-omega-甲氧基 PEG [分子量：2-kDa (PEG(M2))、5-kDa (PEG(M5))] 结合。另一方面，α-琥珀酰亚胺基-omega-甲氧基 PEG [Mw：2-kDa (PEG(S2))、5-kDa (PEG(S5))] 直接与 Lys 残基结合。 PEG 缀合的 HSA-FeXP 的 MALDI-TOF MS 显示出不同的分子离子峰，这提供了 PEG 链的准确数量。对于 PEG(MY)(HSA-FeXP)，硫醇基团的光谱分析还提供了聚合物结合数的平均值，并且修饰程度由 [IMT]/[HSA] 的比率控制。 2-kDa PEG 缀合物（磷酸盐缓冲盐溶液，[HSA] = 5 g dL(-1)）的粘度和胶体渗透压几乎与未修饰的相同，而 5-kDa PEG 结合增加了流变参数。 HSA 表面上柔性聚合物的存在延缓了 O2 与 FeXP 的缔合反应，并稳定了含氧复合物。此外，PEG(MY)(HSA-FeXP) 在大鼠体内表现出较长的 FeXP 循环寿命（13-16 小时）。基于这些结果，可以得出结论：PEG对HSA-FeXP进行表面修饰，提高了其综合输氧能力。特别是，PEG(MY)(HSA-FeXP) 溶液可能是一种很有前途的材料，可用于完全合成的携氧血浆扩张剂作为红细胞替代品。
Artificial O2-carrying hemoprotein composed of human serum albumin including tetrakis(o-amidophenyl)porphinatoiron(II) (Fe4P or Fe3P) [HSA-FeXP] has been modified by maleimide- or succinimide-terminated poly(ethylene glycol) (PEG), and the formed PEG bioconjugates have been physicochemically characterized. 2-Iminothiolane (IMT) reacted with the amino groups of Lys to create active thiol groups, which bind to alpha-maleimide-omega-methoxy PEG [Mw: 2-kDa (PEG(M2)), 5-kDa (PEG(M5))]. On the other hand, alpha-succinimidyl-omega-methoxy PEG [Mw: 2-kDa (PEG(S2)), 5-kDa (PEG(S5))] directly binds to Lys residues. MALDI-TOF MS of the PEG-conjugated HSA-FeXP showed distinct molecular ion peaks, which provide an accurate number of the PEG chains. In the case of PEG(MY)(HSA-FeXP), the spectroscopic assay of the thiol groups also provided the mean of the binding numbers of the polymers, and the degree of the modification was controlled by the ratio of [IMT]/[HSA]. The viscosity and colloid osmotic pressures of the 2-kDa PEG conjugates (phosphate-buffered saline solution, [HSA] = 5 g dL(-1)) were almost the same as that of the nonmodified one, whereas the 5-kDa PEG binding increased the rheological parameters. The presence of flexible polymers on the HSA surface retarded the association reaction of O2 to FeXP and stabilized the oxygenated complex. Furthermore, PEG(MY)(HSA-FeXP) exhibited a long circulation lifetime of FeXP in rats (13-16 h). On the basis of these results, it can be concluded that the surface modification of HSA-FeXP by PEG has improved its comprehensive O2-transporting ability. In particular the PEG(MY)(HSA-FeXP) solution could be a promising material for entirely synthetic O2-carrying plasma expander as a red cell substitute.