Affinity switching for lysozyme and dual-responsive microgels by stopped-flow technique: Kinetic control and activity evaluation
Affinity switching for lysozyme and dual-responsive microgels by stopped-flow technique: Kinetic control and activity evaluation
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通过停流技术进行溶菌酶和双响应微凝胶的亲和力转换：动力学控制和活性评估
DOI：
10.1007/s10118-017-1948-z
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发表时间：
2017-05
影响因子：
4.3
通讯作者：
Yin Jun
中科院分区：
文献类型：
--
作者：
Wu Yun zhu;Zhang Zhi huang;Han Xin;Zhang Jian;Zhang Wen ming;Yin Jun
利用蛋白质作为纳米医学的治疗手段是一个新兴的研究领域，发展迅速。然而，蛋白质在体内总是容易被肾脏排泄或被蛋白质水解系统消化，这在很大程度上限制了它们的使用。尽管生物相容性聚合物已与蛋白质共价连接，以保护它们不被免疫系统识别并延长其循环时间，但它们的生物活性有时会降低。为了填补这一空白，采用了物理隔离、包装或封装技术。到目前为止，已有各种成熟的案例报道，但对客体分子加载和释放的整个时间尺度，特别是初始快速加载过程的研究却很少。本文合成了一系列双响应的聚n -异丙基丙烯酰胺-共甲基丙烯酸（P(NIPAM-co-MAA)）微凝胶，并在停流仪上研究了溶菌酶在外界刺激调节下的络合和释放动力学，该微凝胶适合于快速动态监测。对吸附动力学早期阶段（< 50 s）的密切观察表明，初始微凝胶在~1 s内发生崩溃，当溶菌酶浓度升高时，观察到更快的转变。所有的动态轨迹都可以用双指数函数很好地拟合，表明弛豫时间分别为快（τ1）和慢（τ2）。然后，利用ph响应特性对微凝胶释放结合溶菌酶进行动力学研究，并在溶血微球菌（M. lysodeikticus）细胞悬液中同步测定释放的溶菌酶的活性。通过拟合记录的动态轨迹计算相应的松弛时间（τ）。我们推测这项工作可以为微凝胶纳米载体用于蛋白质的递送、控释和可能的化学分离提供基本的动力学数据和理论依据。
The use of proteins as therapeutics in nanomedicine is an emerging research field and has developed rapidly. However, proteins are always vulnerable to renal excretion or digestion by the proteolytic systemin vivo, which limits their usage to a large extent. Although biocompatible polymers have been covalently linked to proteins to protect them from recognition by the immune system and prolong their circulation time, the biological activity of them is sometimes decreased. To fill this gap, physical isolation, wrapping, or encapsulation techniques are employed. Up to now, various mature examples were reported, but the whole time scales for guest molecules loading and releasing, especially the initial rapid loading process, were rarely mentioned. Herein, a series of dual-responsive poly(N-isopropylacrylamide-co-methacrylic acid) (P(NIPAM-co-MAA)) microgels were synthesized and employed to investigate the kinetics ofin situcomplexation and release of lysozyme under external stimuli modulation upon a stopped-flow apparatus, which was suitable for rapid dynamic monitoring. Close inspection of the adsorption kinetics during the early stages (< 50 s) revealed that the initial microgel collapse occurred within ~1 s, with more rapid transitions being observed when higher lysozyme concentrations were targeted. All the dynamic traces could be well fitted with a double exponential function, suggesting a fast (τ1) and a slow (τ2) relaxation time, respectively. Then, the kinetics of releasing bound lysozyme from microgels was carried on by utilizing the pH-responsive property, and the evaluation of the activity of released lysozyme was synchronously measured in a Micrococcus lysodeikticus (M. lysodeikticus) cell suspension. The corresponding relaxation time (τ) was also calculated by fitting the recorded dynamic traces. We speculate that this work can provide basic dynamics data and theoretical basis for microgels based nanocarriers to be used for protein delivery, controlled release, and possible chemical separation.
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影响因子：
3.3
作者：
Shuiqin Zhou;B. Chu
通讯作者：
Shuiqin Zhou;B. Chu
影响因子：
6.2
作者：
Hong Zhang;Sawitri Mardyani;W. Chan;E. Kumacheva
通讯作者：
Hong Zhang;Sawitri Mardyani;W. Chan;E. Kumacheva
影响因子：
2.5
作者：
Plamper, FA;Becker, H;Müller, AHE
通讯作者：
Müller, AHE
影响因子：
16.6
作者：
Beierle, John M.;Yoshimatsu, Keiichi;Shea, Kenneth J.
通讯作者：
Shea, Kenneth J.
影响因子：
2.1
作者：
Nakimbugwe, D;Masschalck, B;Michiels, CW
通讯作者：
Michiels, CW