В этой статье, высокая пропускная способность метода представлена для синтеза олигосахаридов и их крепление к поверхности наночастицы polyanhydride для дальнейшего использования в ориентированных на конкретные рецепторы на антиген представляющих клеток.
Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design efficacious vaccines carriers. Nanoparticle-based platforms can prolong the persistence of vaccine antigens, which could improve vaccine immunogenicity1. Several biodegradable polymers have been studied as vaccine delivery vehicles1; in particular, polyanhydride particles have demonstrated the ability to provide sustained release of stable protein antigens and to activate antigen presenting cells and modulate immune responses2-12.
The molecular design of these vaccine carriers needs to integrate the rational selection of polymer properties as well as the incorporation of appropriate targeting agents. High throughput automated fabrication of targeting ligands and functionalized particles is a powerful tool that will enhance the ability to study a wide range of properties and will lead to the design of reproducible vaccine delivery devices.
The addition of targeting ligands capable of being recognized by specific receptors on immune cells has been shown to modulate and tailor immune responses10,11,13 C-type lectin receptors (CLRs) are pattern recognition receptors (PRRs) that recognize carbohydrates present on the surface of pathogens. The stimulation of immune cells via CLRs allows for enhanced internalization of antigen and subsequent presentation for further T cell activation14,15. Therefore, carbohydrate molecules play an important role in the study of immune responses; however, the use of these biomolecules often suffers from the lack of availability of structurally well-defined and pure carbohydrates. An automation platform based on iterative solution-phase reactions can enable rapid and controlled synthesis of these synthetically challenging molecules using significantly lower building block quantities than traditional solid-phase methods16,17.
Herein we report a protocol for the automated solution-phase synthesis of oligosaccharides such as mannose-based targeting ligands with fluorous solid-phase extraction for intermediate purification. After development of automated methods to make the carbohydrate-based targeting agent, we describe methods for their attachment on the surface of polyanhydride nanoparticles employing an automated robotic set up operated by LabVIEW as previously described10. Surface functionalization with carbohydrates has shown efficacy in targeting CLRs10,11 and increasing the throughput of the fabrication method to unearth the complexities associated with a multi-parametric system will be of great value (Figure 1a).
Эффективность углеводов ориентации агентов прямых взаимодействий наночастиц на клетки иммунной системы была продемонстрирована ранее 10, 11. Предыдущие исследования в нашей лаборатории показали, что конкретные сахара придает polyanhydride наночастицы способны ориентированы на разли?…
The authors have nothing to disclose.
Авторы хотели бы поблагодарить армии США медицинских исследований и материального Command (грант № W81XWH-10-1-0806) и Национальные институты здравоохранения (грант № U19 AI091031-01 и грант № 1R01GM090280) за финансовую поддержку. Б. признает Balloun профессора в области химической и биологической инженерии и NLBP признает профессора Уилкинсона междисциплинарных технических наук. Мы благодарим Юлию Вела за помощь в проведении экспериментов наночастиц функционализации.
Name | Company | Catalog number |
Motorized XYZ Stage: 3x T-LSM050A, 50 mm travel per axis | Zaber Technologies | T-XYZ-LSM050A-KT04 |
NE-1000 Single Syringe Pump | New Era Pump Systems | NE-1000 |
Pyrex* Vista* Rimless Reusable Glass Culture Tubes | Corning | 07-250-125 |
ASW 1000 | Chemspeed Technologies | |
LabVIEW | National Instruments | 776671-35 |
SGE Gas Tight Syringes, Luer Loc | Sigma Aldrich | 509507 |
XL-2000 Sonicator | Qsonica | Q55 |
Mini-tube rotator | Fisher Scientific | 05-450-127 |