Suleiman Ehsan, Damm Dominik, Batzoni Mirjam, Temchura Vladimir, Wagner Andreas, Überla Klaus, Vorauer-Uhl Karola
Polymun Scientific Immunbiologische Forschung GmbH, 3400 Klosterneuburg, Austria.
Department of Biotechnology, University of Natural Resources and Life Sciences, 1190 Vienna, Austria.
Pharmaceutics. 2019 Nov 18;11(11):619. doi: 10.3390/pharmaceutics11110619.
Since the first use of liposomes as carriers for antigens, much work has been done to elucidate the mechanisms involved in the encapsulation of vaccine-relevant biomolecules. However, only a few studies have specifically investigated the encapsulation of hydrophilic, non-conformational peptide epitopes. We performed comprehensive and systematic screening studies, in order to identify conditions that favor the electrostatic interaction of such peptides with lipid membranes. Moreover, we have explored bi-terminal sequence extension as an approach to modify the isoelectric point of peptides, in order to modulate their membrane binding behavior and eventually shift/expand the working range under which they can be efficiently encapsulated in an electrostatically driven manner. The findings of our membrane interaction studies were then applied to preparing peptide-loaded liposomes. Our results show that the magnitude of membrane binding observed in our exploratory in situ setup translates to corresponding levels of encapsulation efficiency in both of the two most commonly employed methods for the preparation of liposomes, i.e., thin-film hydration and microfluidic mixing. We believe that the methods and findings described in the present studies will be of use to a wide audience and can be applied to address the ongoing relevant issue of the efficient encapsulation of hydrophilic biomolecules.
自从首次将脂质体用作抗原载体以来，人们开展了大量工作以阐明与疫苗相关生物分子包封有关的机制。然而，仅有少数研究专门考察了亲水性、非构象性肽表位的包封情况。我们进行了全面而系统的筛选研究，以确定有利于此类肽与脂质膜发生静电相互作用的条件。此外，我们探索了通过双末端序列延伸来修饰肽的等电点，从而调节其膜结合行为，并最终改变/扩大其能够以静电驱动方式高效包封的工作范围。然后，我们将膜相互作用研究的结果应用于制备载肽脂质体。我们的结果表明，在我们探索性的原位实验装置中观察到的膜结合程度，在两种最常用的脂质体制备方法（即薄膜水化法和微流混合法）中都转化为相应水平的包封效率。我们相信，本研究中描述的方法和发现将对广大读者有用，并且可用于解决当前有关亲水性生物分子高效包封的相关问题。
