Zhao Guangzu, Azuar Armira, Toth Istvan, Skwarczynski Mariusz
School of Chemistry and Molecular Biosciences, The University of Queensland, St Lucia, Australia.
Institute for Molecular Bioscience, The University of Queensland, St Lucia, Australia.
Bio Protoc. 2021 Apr 5;11(7):e3973. doi: 10.21769/BioProtoc.3973.
Most vaccines require co-delivery of an adjuvant in order to generate the desired immune responses. However, many currently available adjuvants are non-biodegradable, have limited efficacy, and/or poor safety profile. Thus, new adjuvants, or self-adjuvanting vaccine delivery systems, are required. Here, we proposed a self-adjuvanting delivery system that is fully defined, biodegradable, and non-toxic. The system is produced by conjugation of polyleucine to peptide antigen, followed by self-assembly of the conjugate into nanoparticles. The protocol includes solid-phase peptide synthesis of the vaccine conjugate, purification, self-assembly and physicochemical characterization of the product. Overall, this protocol describes, in detail, the production of a well-defined and effective self-adjuvanting delivery system for peptide antigens, along with tips for troubleshooting.
大多数疫苗需要共同递送佐剂以产生所需的免疫反应。然而，许多目前可用的佐剂不可生物降解，效力有限，和/或安全性差。因此，需要新的佐剂或自佐剂疫苗递送系统。在此，我们提出了一种完全确定、可生物降解且无毒的自佐剂递送系统。该系统通过将聚亮氨酸与肽抗原偶联，然后将偶联物自组装成纳米颗粒来制备。该方案包括疫苗偶联物的固相肽合成、纯化、产物的自组装和理化表征。总体而言，该方案详细描述了一种用于肽抗原的定义明确且有效的自佐剂递送系统的生产方法，以及故障排除提示。
