a Department of Chemical Engineering , Villanova University , Villanova , PA , USA.
b Division of Hematology , The Children's Hospital of Philadelphia , Philadelphia , PA , USA.
Artif Cells Nanomed Biotechnol. 2018;46(sup2):1137-1144. doi: 10.1080/21691401.2018.1480491. Epub 2018 Jun 19.
Since donated red blood cells must be constantly refrigerated, they are often unavailable in remote areas and battlefields. The goal of this study was to synthesize a highly stable blood substitute that does not require refrigeration. Specifically, the extracellular haemoglobin (a.k.a. erythrocruorin, Ec) of the earthworm Lumbricus terrestris erythrocruororin (LtEc) was cross-linked with poly(acrylic acid) (PAA) and ethylene diamine (EDA). PAGE analysis of the LtEc nanoparticles reveals cross-linking between subunits, while dynamic light scattering and scanning electron microscopy show that cross-linking significantly increases the size of the LtEc nanoparticles (164 ± 13.9 nm). Cross-linking also significantly increased the thermal stability of the LtEc nanoparticles by 10 °C (T = 72 ± 0.84 °C) relative to native LtEc (T = 62 ± 0.6 °C). In addition, while native LtEc rapidly dissociates at pH 9, the LtEc nanoparticles resist subunit dissociation up to pH 10. The oxygen affinity of the LtEc nanoparticles (P = 6.85 ± 0.13 mm Hg) is much higher than native LtEc (P = 26.67 ± 0.4 mm Hg), but the cooperativity (n = 2.43 ± 0.12) is not affected. Altogether, these results show that cross-linking LtEc with PAA and EDA provides a potential blood substitute with increased stability and oxygen affinity.
由于捐赠的红细胞必须不断冷藏，因此它们在偏远地区和战场上往往无法获得。本研究的目的是合成一种不需要冷藏的高度稳定的血液替代品。具体来说，通过将蚯蚓血影蛋白（即血影红蛋白，Ec）与聚丙烯酸（PAA）和乙二胺（EDA）交联，来合成血影蛋白纳米颗粒。聚丙烯酰胺凝胶电泳（PAGE）分析表明 Ec 亚基之间发生了交联，而动态光散射和扫描电子显微镜显示交联显著增加了 LtEc 纳米颗粒的尺寸（164 ± 13.9nm）。交联还使 LtEc 纳米颗粒的热稳定性显著提高了 10°C（T=72 ± 0.84°C），而天然 LtEc 的热稳定性仅提高了 6°C（T=62 ± 0.6°C）。此外，虽然天然 LtEc 在 pH 9 时迅速解离，但 LtEc 纳米颗粒在 pH 10 时仍能抵抗亚基解离。LtEc 纳米颗粒的氧亲和力（P=6.85 ± 0.13mmHg）远高于天然 LtEc（P=26.67 ± 0.4mmHg），但其协同性（n=2.43 ± 0.12）不受影响。总的来说，这些结果表明，用 PAA 和 EDA 交联 LtEc 提供了一种具有更高稳定性和氧亲和力的潜在血液替代品。
