Comprehensive characterization of tense and relaxed quaternary state glutaraldehyde polymerized bovine hemoglobin as a function of cross-link density.,Biotechnology and Bioengineering

当前位置： X-MOL 学术 › Biotechnol. Bioeng. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Comprehensive characterization of tense and relaxed quaternary state glutaraldehyde polymerized bovine hemoglobin as a function of cross-link density.
Biotechnology and Bioengineering ( IF 3.5 ) Pub Date : 2020-05-30 , DOI: 10.1002/bit.27382
Xiangming Gu ₁ , Crystal Bolden-Rush ₁ , Clayton T Cuddington ₁ , Donald A Belcher ₁ , Chintan Savla ₁ , Ivan S Pires ₁ , Andre F Palmer ₁

Affiliation

Previously, our lab developed high molecular weight (MW) tense (T) quaternary state glutaraldehyde polymerized bovine hemoglobins (PolybHbs) that exhibited reduced vasoactivity in several small animal models. In this study, we prepared PolybHb in the T and relaxed (R) quaternary state with ultrahigh MW (>500 kDa) with varying cross‐link densities, and investigated the effect of MW on key biophysical properties (i.e., O₂ affinity, cooperativity (Hill) coefficient, hydrodynamic diameter, polydispersity, polymer composition, viscosity, gaseous ligand‐binding kinetics, auto‐oxidation, and haptoglobin [Hp]‐binding kinetics). To further optimize current PolybHb synthesis and purification protocols, we performed a comprehensive meta‐data analysis to evaluate correlations between procedural parameters (i.e., cross‐linker:bovine hemoglobin (bHb) molar ratio, gas‐liquid exchange time, temperature during sodium dithionite addition, and number of diafiltration cycles) and the biophysical properties of both T‐ and R‐state PolybHbs. Our results showed that, the duration of the fast‐step auto‐oxidation phase of R‐state PolybHb increased with decreasing glutaraldehyde:bHb molar ratio. Additionally, T‐state PolybHbs exhibited significantly higher bimolecular rate constants for binding to Hp and unimolecular O₂ offloading rate constants compared to R‐state PolybHbs. The methemoglobin (metHb) level in the final product was insensitive to the molar ratio of glutaraldehyde to bHb for all PolybHbs. During tangential flow filtration processing of the final product, 14 diafiltration cycles was found to yield the lowest metHb level.

中文翻译：

作为交联密度函数的紧张和松弛四元戊二醛聚合牛血红蛋白的综合表征。

以前，我们的实验室开发了高分子量 (MW) 时态 (T) 季戊二醛聚合牛血红蛋白 (PolybHbs)，其在几种小动物模型中表现出降低的血管活性。在这项研究中，我们制备了具有不同交联密度的超高分子量（>500 kDa）的 T 和松弛 (R) 四元状态的 PolybHb，并研究了 MW 对关键生物物理特性（即 O ₂亲和性、协同性 (Hill) 系数、流体动力学直径、多分散性、聚合物组成、粘度、气态配体结合动力学、自氧化和触珠蛋白 [Hp] 结合动力学）。为了进一步优化当前的 PolybHb 合成和纯化方案，我们进行了全面的元数据分析以评估程序参数（即交联剂：牛血红蛋白 (bHb) 摩尔比、气液交换时间、连二亚硫酸钠添加过程中的温度）之间的相关性和渗滤循环次数）以及 T 态和 R 态 PolybHbs 的生物物理特性。我们的结果表明，随着戊二醛：bHb 摩尔比的降低，R 态 PolybHb 的快速自动氧化阶段的持续时间增加。此外，_{2 个}卸载速率常数与 R 态 PolybHbs 相比。对于所有 PolybHb，最终产品中的高铁血红蛋白 (metHb) 水平对戊二醛与 bHb 的摩尔比不敏感。在最终产品的切向流过滤加工过程中，发现 14 个渗滤循环产生最低的甲基化血红蛋白水平。

更新日期：2020-07-13

点击分享查看原文

点击收藏

公开下载

阅读更多本刊最新论文本刊介绍/投稿指南11