Name: combinatorial synthesis of and high-throughput protein release from polymer film and nanoparticle libraries
Uploaded: 2012-09-06T12:00:00
Description: Watch this Scientific Journal Video about Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries at JoVE.com

Forfatterne erkjenner ONR-MURI Award (NN00014-06-1-1176) og US Army Medical Research og forsyningskommando Command (Grant No W81XWH-10-1-0806) for økonomisk støtte. Dette materialet er basert på arbeid støttet av National Science Foundation i henhold Grant No EEC 0552584 og 0851519.

1. Kombinatorisk Polymer Library Synthesis (Varierende i Polymerkjemi) - se figur 1 for Robotic Setup <ol><li> Oppløs hver monomer i passende oppløsningsmiddel (konsentrasjon = 25 mg / ml) og laste hver inn i en 10cc gasstett sprøyte. </li><li> Fest løsemiddelbestandig lokke lås kapillarrør til slutten av hver sprøyte. </li><li> Plasser sprøyter på sprøyten pumper (New Era Programmerbare sprøyte pumper) og låses i posisjon. </li><li> Still lineære aktuatorer (Zaber) til startposisjon. </l...

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	<li>Petersen, L. K., Sackett, C. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+novel,+high-throughput+method+to+study+in+vitro+protein+release+from+polymer+nanospheres.">A novel, high-throughput method to study in vitro protein release from polymer nanospheres.</a> J. Comb. Chem. 12, 51-56 (2010).</li><li>Petersen, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Activation+of+innate+immune+responses+in+a+pathogen-mimicking+manner+by+amphiphilic+polyanhydride+nanoparticle+adjuvants.">Activation of innate immune responses in a pathogen-mimicking manner by amphiphilic polyanhydride nanoparticle adjuvants.</a> Biomaterials. 32, 6815-6822 (2011).</li><li>Vogel, B. M., Cabral, J. T., Eidelman, N., Narasimhan, B., Mallapragada, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Parallel+synthesis+and+high-throughput+dissolution+testing+of+biodegradable+polyanhydride+copolymers.">Parallel synthesis and high-throughput dissolution testing of biodegradable polyanhydride copolymers.</a> J. Comb. Chem. 7, 921-928 (2005).</li><li>Petersen, L. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+evaluation+of+in+vivo+biodistribution+of+polyanhydride+nanoparticles.">High-throughput evaluation of in vivo biodistribution of polyanhydride nanoparticles.</a> Adv. Healthcare Mater. , (2012).</li><li>Petersen, L. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combinatorial+design+of+biomaterials+for+drug+delivery:+opportunities+and+challenges.">Combinatorial design of biomaterials for drug delivery: opportunities and challenges.</a> Expert Opin. Drug Deliv. 5, 837-846 (2008).</li><li>Petersen, L. K., Oh, J., Sakaguchi, D. S., Mallapragada, S. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amphiphilic+polyanhydride+films+promote+neural+stem+cell+adhesion+and+differentiation.">Amphiphilic polyanhydride films promote neural stem cell adhesion and differentiation.</a> Tissue Eng. 17, 2533-2541 (2011).</li><li>Petersen, L. K., Sackett, C. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+analysis+of+protein+stability+in+polyanhydride+nanoparticles.">High-throughput analysis of protein stability in polyanhydride nanoparticles.</a> Acta Biomater. 6, 3873-3881 (2010).</li><li>Petersen, L. K., Xue, L., Wannemuehler, M. J., Rajan, K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+simultaneous+effect+of+polymer+chemistry+and+device+geometry+on+the+in+vitro+activation+of+murine+dendritic+cells.">The simultaneous effect of polymer chemistry and device geometry on the in vitro activation of murine dendritic cells.</a> Biomaterials. 30, 5131-5142 (2009).</li><li>Thorstenson, J. B., Petersen, L. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Combinatorial/high-throughput+methods+for+the+determination+of+polyanhydride+phase+behavior.">Combinatorial/high-throughput methods for the determination of polyanhydride phase behavior.</a> J. Comb. Chem. 11, 820-828 (2009).</li><li>Xue, L., Petersen, L., Broderick, S., Narasimhan, B., Rajan, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identifying+factors+controlling+protein+release+from+combinatorial+biomaterial+libraries+via+hybrid+data+mining+methods.">Identifying factors controlling protein release from combinatorial biomaterial libraries via hybrid data mining methods.</a> ACS Comb. Sci. 13, 50-58 (2011).</li><li>Adler, A. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-throughput+cell-based+screening+of+biodegradable+polyanhydride+libraries.">High-throughput cell-based screening of biodegradable polyanhydride libraries.</a> Comb. Chem. High Through. Screen. 12, 634-645 (2009).</li><li>Determan, A. S., Trewyn, B. G., Lin, V. S., Nilsen-Hamilton, M., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encapsulation,+stabilization,+and+release+of+BSA-FITC+from+polyanhydride+microspheres.">Encapsulation, stabilization, and release of BSA-FITC from polyanhydride microspheres.</a> J. Control. Release. 100, 97-109 (2004).</li><li>Determan, A. S., Wilson, J. H., Kipper, M. J., Wannemuehler, M. J., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein+stability+in+the+presence+of+polymer+degradation+products:+consequences+for+controlled+release+formulations.">Protein stability in the presence of polymer degradation products: consequences for controlled release formulations.</a> Biomaterials. 27, 3312-3320 (2006).</li><li>Torres, M. P., Determan, A. S., Anderson, G. L., Mallapragada, S. K., Narasimhan, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Amphiphilic+polyanhydrides+for+protein+stabilization+and+release.">Amphiphilic polyanhydrides for protein stabilization and release.</a> Biomaterials. 28, 108-116 (2007).</li><li>Torres, M. P., Vogel, B. M., Narasimhan, B., Mallapragada, S. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+characterization+of+novel+polyanhydrides+with+tailored+erosion+mechanisms.">Synthesis and characterization of novel polyanhydrides with tailored erosion mechanisms.</a> J. Biomed. Mater. Res. A. 76, 102-110 (2006).</li><li>Carrillo-Conde, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Encapsulation+into+amphiphilic+polyanhydride+microparticles+stabilizes+Yersinia+pestis+antigens.">Encapsulation into amphiphilic polyanhydride microparticles stabilizes Yersinia pestis antigens.</a> Acta Biomater. 6, 3110-3119 (2010).</li></ol>

Kjennskap nødvendige syntese forhold og glass omvandlingstemperaturer (T g) av polymerer blir syntetisert er avgjørende for biblioteket fabrikasjon. Dersom T g s er under romtemperatur, kan nanopartikkel fabrikasjon trinnet må utføres i en kontrollert temperatur miljø under T g av polymerene. I tillegg bør forsiktighet tas for å sikre at alt utstyr som kommer i kontakt med høye temperaturer og løsemidler må være skikket til å håndtere disse forholdene. Flere av parametrene i...

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combinatorial synthesis of and high-throughput protein release from polymer film and nanoparticle libraries

Polyanhydrides er en klasse av biomaterialer med vevsvennligheten og levering av legemidler evner. Mens de har blitt studert med konvensjonell en-utvalgs-at-a-time syntese teknikker, har en nyere high-throughput tilnærming blitt utviklet slik at syntese og testing av store biblioteker av polyanhydrides en. Dette vil lette mer effektiv optimalisering og designprosessen av disse biomaterialer for narkotika og vaksine levering applikasjoner. Metoden i dette arbeidet beskriver kombinatoriske syntese av biologisk nedbrytbart polyanhydride film og nanopartikkel biblioteker og high-throughput påvisning av protein utgivelse fra disse bibliotekene. I denne robotically operert metode (figur 1), er lineære aktuatorer og sprøyte pumper kontrollert av LabVIEW, som muliggjør en hands-free automatisert protokollen, eliminere brukerfeil. Videre gjør denne metoden den raske fabrikasjon av mikroskala polymer biblioteker, røducing batch størrelse mens resulterte i etableringen av multivariant polymer systemer. Dette kombinatoriske tilnærming til polymer syntese forenkler syntese av opptil 15 forskjellige polymerer i en tilsvarende mengde tid det ville ta å syntetisere en polymer konvensjonelt. I tillegg kan polymer kombinatoriske biblioteket bli fabrikkert i blanke eller protein-loaded geometrier inkludert filmer eller nanopartikler Ved oppløsning av polymeren biblioteket i et løsningsmiddel og nedbør inn i en ikke-oppløsningsmiddel (for nanopartikler) eller ved vakuumtørking (for filmer). Etter lasting en fluorokrom-konjugert protein inn i polymeren bibliotekene kan protein frigivelse kinetikk vurderes ved høy gjennomstrømning ved hjelp av en fluorescens-basert deteksjonsmetode (figurene 2 og 3) som beskrevet tidligere 1. Dette kombinatoriske plattformen har blitt validert med konvensjonelle metoder 2 og polyanhydride film og nanopartikkel bibliotekene har vært preget med <sup&gt; 1 H NMR og FTIR. Bibliotekene har blitt vist for protein utgivelsen kinetikk, stabilitet og antigenisitet, in vitro cellulær toksisitet, cytokine produksjon, overflate markør uttrykk, heft, spredning og differensiering, og in vivo biodistribusjon og mucoadhesion 1-11. Den kombinatoriske metode utviklet heri muliggjør high-throughput polymer syntese og produksjon av protein-loaded nanopartikler og film biblioteker, som kan, i sin tur bli vist in vitro og in vivo for optimalisering av biomateriale ytelse.

Watch this Scientific Journal Video about Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries at JoVE.com

Combinatorial Synthesis of and High-throughput Protein Release from Polymer Film and Nanoparticle Libraries

Denne metoden beskriver kombinatoriske syntese av biologisk nedbrytbart polyanhydride film og nanopartikkel biblioteker og high-throughput påvisning av protein utgivelse fra disse bibliotekene.

Kombinatorisk Syntese av og High-throughput Protein År fra Polymer Film og nanopartikler Libraries

Polyanhydrides are a class of biomaterials with excellent biocompatibility and drug delivery capabilities. While they have been studied extensively with ...

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