कार्बोहाइड्रेट के संश्लेषण उच्च throughput और Polyanhydride नैनोकणों functionalization

Summary

इस अनुच्छेद में, एक उच्च throughput विधि oligosaccharides और उनके लगाव की प्रतिजन कोशिकाओं पर विशेष रिसेप्टर्स लक्ष्यीकरण में आगे उपयोग के लिए polyanhydride नैनोकणों के सतह के संश्लेषण के लिए प्रस्तुत किया है.

Abstract

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 responses^2-12.

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 responses^10,11,13 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 activation^14,15. 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 methods^16,17.

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 CLRs^10,11 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).

Protocol

1. उच्च throughput कार्बोहाइड्रेट संश्लेषण स्वचालित dimannoside का संश्लेषण, एक उपयुक्त संरक्षित चीनी दाता, आम तौर पर trichloroacetimidate, और स्वीकर्ता, मुख्य रूप से एक alkenyl fluorous शराब, पहले बेंच शीर्ष पर संश्लेषित कर रहे हैं. …

Discussion

प्रत्यक्ष nanoparticle बातचीत करने के लिए एजेंटों प्रतिरक्षा कोशिकाओं को लक्षित के रूप में कार्बोहाइड्रेट की प्रभावकारिता पहले किया गया है ^{11 10,} प्रदर्शन. हमारे प्रयोगशालाओं में पिछले अनुसंधान दिखाया है …

Materials

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