使用表面巯基化牛血清白蛋白和 Traut's 试剂通过简便的超声化学合成法制备壳稳定的反应性微泡。
A facile sonochemical synthesis of shell-stabilized reactive microbubbles using surface-thiolated bovine serum albumin with the Traut's reagent.
机构信息
Department of Chemical and Materials Engineering, Donadeo Innovation Centre for Engineering, University of Alberta, 9211 - 116 St NW, Edmonton T6G 1H9, Canada.
Department of Chemical and Materials Engineering, Donadeo Innovation Centre for Engineering, University of Alberta, 9211 - 116 St NW, Edmonton T6G 1H9, Canada.
出版信息
Ultrason Sonochem. 2017 May;36:454-465. doi: 10.1016/j.ultsonch.2016.12.033. Epub 2016 Dec 26.
The short lifetime of proteinaceous microbubbles produced using conventional sonication method has hindered their applications in drug delivery and metal removal from wastewater. In this study, we aimed to synthesize stable proteinaceous microbubbles and to demonstrate their reactivity. Our model protein, bovine serum albumin (BSA) was treated with 2-iminothiolane hydrochloride (Traut's reagent) to convert primary amines to thiols before the synthesis of microbubbles. Microbubbles produced with the Traut's reagent-treated BSA (BSA-SH MBs) were initially concentrated at median sizes of 0.5 and 2.5μm. The 0.5μm portion quickly vanished, and the 2.5μm portion gradually shrank to ∼850nm in ∼3days and became stabilized afterward for several months under 4°C. Characterizations of BSA-SH MBs by Fourier transform infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS) indicated the presence of free unbound thiols and primary amines on their surface, implying the possibility of further surface modification. Based on the zeta potential measurement, the isoelectric point (IEP) of BSA-SH MBs was determined to be 4.5. The attachments of BSA-SH MBs on alumina, silica, and gold surfaces in different pH environments were carried out with a quartz crystal microbalance with dissipation monitoring (QCM-D), demonstrating the reactivities of BSA-SH MBs. At pH 6, the negatively charged BSA-SH MBs were adsorbed onto the alumina surface by electrostatic interaction. Analogously, at pH 4, the adsorption of the positively charged BSA-SH MBs on the silica surface was confirmed. Compared with the electrostatic interaction, the adsorption of BSA-SH MBs on the gold surface is attributed to the strong gold-thiol bonding effect. This is the first time that a universal approach for stabilizing protein-shelled microbubbles was reported using only one single step of surface treatment of proteins with the Traut's reagent.
传统超声法制备的蛋白质微泡的短寿命限制了其在药物输送和从废水中去除金属等方面的应用。在本研究中，我们旨在合成稳定的蛋白质微泡并证明其反应性。我们的模型蛋白牛血清白蛋白（BSA）先用 2-亚氨基硫代盐酸盐（Traut 试剂）处理，将伯胺转化为硫醇，然后再合成微泡。用 Traut 试剂处理的 BSA（BSA-SH MBs）制备的微泡最初在 0.5 和 2.5μm 的中值尺寸处浓缩。0.5μm 部分迅速消失，而 2.5μm 部分在大约 3 天内逐渐收缩到约 850nm，此后在 4°C 下稳定几个月。傅里叶变换红外（FTIR）光谱和 X 射线光电子能谱（XPS）对 BSA-SH MBs 的表征表明，其表面存在游离的未结合的硫醇和伯胺，这意味着可以进一步进行表面修饰。根据zeta 电位测量，BSA-SH MBs 的等电点（IEP）确定为 4.5。在不同 pH 环境下，用石英晶体微天平（QCM-D）进行了 BSA-SH MBs 在氧化铝、二氧化硅和金表面的附着实验，证明了 BSA-SH MBs 的反应性。在 pH6 下，带负电荷的 BSA-SH MBs 通过静电相互作用吸附在氧化铝表面上。类似地，在 pH4 下，证实了带正电荷的 BSA-SH MBs 吸附在二氧化硅表面上。与静电相互作用相比，BSA-SH MBs 在金表面的吸附归因于强的金-硫键合效应。这是首次报道仅用 Traut 试剂对蛋白质进行一步表面处理，就可以稳定蛋白质壳微泡的通用方法。
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