Name: high-throughput synthesis of carbohydrates and functionalization of polyanhydride nanoparticles
Uploaded: 2012-07-06T12:00:00
Description: Watch this Scientific Journal Video about High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles at JoVE.com

Forfatterne vil gerne takke den amerikanske hær Medical Research og Materiel Kommando (Grant # W81XWH-10-1-0806) og National Institutes of Health (Grant # U19 AI091031-01 og Grant # 1R01GM090280) for finansiel støtte. BN anerkender den Balloun professoratet i kemiske og biologiske Engineering og NLBP anerkender Wilkinson professor i Interdisciplinary Engineering. Vi takker Julia Vela for sin bistand til gennemførelsen af ​​nanopartikel funktionalisering eksperimenter.

1. High-throughput Carbohydrate Syntese <ol><li> Før den automatiserede syntese af dimannoside, et passende beskyttet sukker donor, typisk trichloracetimidat og acceptor, fortrinsvis en alkenylgruppe fluorous alkohol, syntetiseres på bench-top. </li><li> Et program skrevet til automatiseret syntese af dimannoside. En skematisk fremstilling af den grundlæggende automatiseret procedure er vist i figur 2. I programmet, sikres det, at før tilsætning af promotoren, er blandingen af ​​donor-og acce...

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Effekten af kulhydrater som målrettende midler til direkte nanopartikel interaktioner til immunceller er tidligere blevet demonstreret 10, 11. Tidligere forskning i vore laboratorier har vist, at specifikke sukkere knyttet til polyanhydrid nanopartikler stand til at målrette forskellige CLR på antigenpræsenterende celler (APC&#39;er) og dermed forbedre aktivering af immunceller, som kan være vigtige for yderligere T-celleaktivering 10, 11. Men for at opnå optimal målrette flere parametre, s?...

<table border="1"><tbody><tr><td> Navn </td><td> Firma </td><td> Katalog nr. </td></tr><tr><td> Motoriseret XYZ Stage: 3x T-LSM050A, 50 mm vandring pr akse </td><td> Zaber Technologies </td><td> T-XYZ-LSM050A-KT04 </td></tr><tr><td> NE-1000 Single sprøjtepumpe </td><td> New Era Pumpesystemer </td><td> NE-1000 </td></tr><tr><td> Pyrex * Vista * Uindfattede Genanvendelige Glas Kultur Tubes </td><td> Corning </td><td> 07-250-125 </td></tr><tr><td> ASW 1000 </td><td> Chemspeed Technologies </td><td></td></tr><tr><td> LabVIEW </td><td> National Instruments </td><td> 776671-35 </td></tr><tr><td> SGE gastætte Sprøjter, Luer Loc </td><td> Sigma Aldrich </td><td> 509507 </td></tr><tr><td> XL-2000 Sonicator </td><td> Qsonica </td><td> Q55 </td></tr><tr><td> Mini-rør rotator </td><td> Fisher Scientific </td><td> 05-450-127 </td></tr></tbody></table>

high-throughput synthesis of carbohydrates and functionalization of polyanhydride nanoparticles

Tværfaglig tilgang med inddragelse af områder som materielt design, nanoteknologi, kemi og immunologi skal udnyttes til rationelt at designe effektive vacciner luftfartsselskaber. Nanopartikel-baserede platforme kan forlænge eksistensen af vaccineantigener, hvilket kan forbedre vaccinens immunogenicitet 1. Flere biologisk nedbrydelige polymerer har været undersøgt som vaccine leveringsvehikler 1, i særdeleshed, har polyanhydrid partikler demonstreret evne til at tilvejebringe langvarig frigivelse af stabile protein-antigener, og at aktivere antigenpræsenterende celler og modulere immunreaktioner 2-12. Den molekylære konstruktion af disse vaccine bærere må integrere en rationel udvælgelse af polymere egenskaber såvel som inkorporering af passende målrettende midler. Høj gennemløb automatiseret fremstilling af målretning ligander og funktionaliserede partikler er et kraftfuldt værktøj, der vil øge muligheden for at studere en bred range af egenskaber og vil føre til udformningen af ​​reproducerbare vaccine tilførselsanordninger. Tilsætningen af målretning ligander, der kan genkendes af specifikke receptorer på immunceller er blevet vist at modulere og skræddersy immunresponser 10,11,13 C-type lectin receptorer (CLR) er mønstergenkendelse receptorer (PRRS), som genkender kulhydrater til stede på overflade af patogener. Stimulering af immunceller via CLR tillader forøget internalisering af antigen og efterfølgende præsentation for yderligere T-celleaktivering 14,15. Derfor carbohydratmolekyler spiller en vigtig rolle i studiet af immunresponser, men anvendelsen af ​​disse biomolekyler ofte lider af manglende tilgængelighed af strukturelt veldefinerede og rene kulhydrater. En automatisering platform baseret på iterativ løsning-fase reaktioner kan aktivere hurtig og kontrolleret syntese af disse syntetisk udfordrende molekyler ved hjælp af betydeligt lavere bPBYGNING blok mængder end traditionelle fastfase-metoder 16,17. Heri vi rapportere en protokol for automatiseret løsning-fase syntese af oligosaccharider, såsom mannose-baserede rettet ligander med fluorous fast-fase-ekstraktion for mellemliggende rensning. Efter udvikling af automatiserede metoder til at gøre det kulhydrat-baserede targetingmiddel, beskriver vi fremgangsmåder til deres fastgørelse på overfladen af polyanhydrid nanopartikler anvendelse af et automatiseret robot, der er nedsat drives LabVIEW som tidligere beskrevet 10. Overflade funktionalisering med kulhydrater har vist effekt i at målrette CLR 10,11 og øge gennemløb af fabrikation metode til at afdække kompleksiteten forbundet med en multi-parametrisk system vil være af stor værdi (figur 1a).

Watch this Scientific Journal Video about High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles at JoVE.com

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

I denne artikel er en high throughput fremgangsmåde præsenteres til syntese af oligosaccharider og deres fastgørelse til overfladen af ​​polyanhydrid nanopartikler til yderligere anvendelse i målretning specifikke receptorer på antigenpræsenterende celler.

High-throughput Syntese af kulhydrater og Funktionalisering af polyanhydridet Nanopartikler

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design ...

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