Yazarlar, ABD Ordusu Tıbbi Araştırma ve Malzeme Komutanlığı teşekkür (Grant # W81XWH-10-1-0806) ve mali destek için Ulusal Sağlık Enstitüleri (Grant # U19 AI091031-01 ve Grant # 1R01GM090280) olacaktır. BN Kimya ve Biyoloji Mühendisliği Balloun Profesörlük kabul eder ve NLBP Disiplinlerarası Mühendislik Wilkinson Profesörlük kabul eder. Biz nanoparçacık fonksiyonlandırmalar deneyler ona yardım için Julia Vela teşekkür ederim.

1. Yüksek verim Karbonhidrat Sentez <ol><li> Önce dimannoside ve otomatik sentezi, uygun bir şekilde korunan şeker donör, genellikle trichloroacetimidate, ve alıcı, özellikle bir alkenil fluorous alkol için tezgah-üst sentezlenir. </li><li> Bir program dimannoside arasında otomatik sentezi için yazılmıştır. Temel otomatikleştirilmiş prosedürün bir şematik Şekil 2 &#39;de gösterilmiştir. Program, bu promotör eklenmesinden önce, verici ve alıcı bir karışımı, en azından 30 dakika süreyle karıştırıldı sağlanır. </li><li> Sentetik donör, alıcı ve trimethylsilyltrifluoromethanesulfonate çözeltileri diklorometan içinde yapılır. Toluen ve diklorometan glikozilasyon reaksiyonlar için, en sık kullanılmaktadır. </li><li> Ayrıca,% 80 metanol ve% 100 metanol içindeki geçici koruyucu gruplarının korumasının kaldırılması için reaktifler çözeltileri hazırlanır. </li><li> Programın başlamasından önce, emin relatOdadaki ive nem otomasyonu odacık içinde% 30 ya da daha düşüktür. Yüksek nem glikozilasyon reaksiyonlar için zararlıdır. </li><li> Program başladıktan sonra, robot kol sırayla reaksiyon şişeye donör ve alıcı çözümleri aktarır. Daha sonra karışım, 30 dakika boyunca karıştırıldı. </li><li> Robotik kol karışımı içine trimethylsilyltrifluoromethanesulfonate arasında 0,2-0,3 benzerleri aktarır Sonra, tipik olarak oda sıcaklığında, -20 gibi düşük sıcaklıklarda ° C de elde edilebilir. Reaksiyon karışımı 30 dakika boyunca karıştırıldı. </li><li> 30 dk sonra reaksiyon durduruldu ve küçük bir alikotu reaksiyon ilerlemesini izlemek için çıkarılır. Tamamlanmaz, reaksiyon devam edilebilir ve sonuçta gerekli süreyi modifiye edilebilir. </li><li> Reaksiyon tamamlandıktan sonra, reaksiyon karışımı saflaştırılması için C&#39;de 8 F 17-değiştirilmiş silika jel içeren fluorous katı faz ekstraksiyonu (FSPE) kartuşlar transfer edilir. </li><li> Sepetisırtlar birinci non-fluorous fraksiyonu kurtulmak için% 80 metanol-su karışımı (8 mL) ile yıkanır. </li><li> Sonra kartuşlar istenen fluorous-etiketli ürünü elde etmek için% 100 metanol ile yıkanır. Ek saflaştırma isteniyorsa, makinenin durduruldu ve reaksiyon ürünü (ler) ilave yoluyla saflaştırma için çıkarılabilir. </li><li> Arıtma döngüsü sonra, robot kol reaksiyon şişeye sodyum metoksid saçıyor. Reaksiyon 2 saat boyunca karıştırıldı. Tam edilmediğinde, reaksiyon yeniden daha uzun bir süre boyunca devam edilebilir ve sonuçta gerekli programlanmış bir süre modifiye edilebilir. </li><li> Reaksiyonun tamamlanmasından sonra, ürün FSPE ile saflaştırılır ve daha sonra kalan suyu uzaklaştırmak için ardından buharlaştırma ile susuz tolüen içinde çözünmesi tabi tutuldu. </li><li> İstenen zincir uzunluğu hedef molekül elde edilir kadar daha sonra çevrimi (adım 6-13 arası) tekrar edilir. </li><li> Otomasyonu elde edilen ürün th korumalı olduğuen fazla saflaştırılmış ve tamamen gibi nükleer manyetik rezonans (NMR) spektroskopisi gibi teknikler ile karakterize edilmektedir. Genellikle patlayıcı hidrojen gazı ve paladyum içerir, çünkü son hedef moleküle korumasının giderilmesinden tam (tüm kalan koruyucu grubun çıkartılması) daha sonra, bir kural olarak otomasyon platformu dışında tamamlanır. Nihai korunmasına son verme işlemi otomasyon platformu dışında tezgah üstü üzerinde gerçekleştirildi. Birinci aşama, bir karboksilik asit ile üretilen aldehit oksidasyonu ile takip fluorous etiketi çift bağın ozonlanmasından idi. Ürün kolon kromatografi ile saflaştırıldı. Son adım paladyum katalizli hidrojenasyon ile benzil eter gruplarının korumasının kaldırılması idi. Ürün, saf nihai ürün elde etmek için paladyum kurtulmak için selit yastığı içinden geçirilmiştir. </li></ol> 2. Yüksek verim Nanopartikül Yüzey fonksiyonlandırmalar <ol><li> Yüksek verim polimer sentezi ve nanoparçacık fabrikasyon aynı pr takip yapılırotocol ve robotik kadar Petersen ve ark 19 tarafından tarif ayarlanır. partikül imalatı için kullanılan kopolimer sistemleri sebasik asit (SA) ve 1,6-bis (para-karboksifenoksi) heksan (CPH), ve 1,8-bis (dayanmaktadır para-karboksifenoksi) -3,6-dioxaoctane (CPTEG) ve CPH. Kullanılmıştır robotik yerleştirme tertibatı şematik temsili Şekil 1b sunulmuştur. </li><li> Nanoparçacık fabrikasyon ardından, nanoparçacık kütüphane ile tüpler içeren tutucu lineer aktüatör sahneye tutturulur. </li><li> Polianhidrid parçacıklarının yüzeyine karbonhidrat tutturulması için, iki ardışık reaksiyon oluşan bir amin-karboksilik asit bağlama reaksiyonu 20 gerçekleştirilir. </li><li> Birinci reaksiyon için, ilk programlanabilir şırınga pompası şırınga 1-etil-3-(3-dimetilaminopropil) ve benzerleri 10 (denklem) (parçacığın yüzey üzerinde ortalama mol karboksilik asit konsantrasyonunun eşdeğer) ile doldurulmuştur-carbodiimide hidroklorür (EDC) ve 10 eq. bir sulu çözelti içinde etilendiaminin, ikinci programlanabilir şırınga pompası şırınganın 12 eq ile yüklenir iken. sulu çözelti içinde N-hidroksisüksinimid (NHS). </li><li> LabVIEW programı kullanarak, reaktif süspansiyonlar nanoparçacık kütüphane * yatırılır. </li><li> Sonra, her bir numune sonicated edilir (40 Hz 30 s) ve tüp tutucu robot platformu kopar. </li><li> Nanoparçacık süspansiyonlar 4 ° C&#39;de sabit dönüş ile 9 saat süreyle inkübe edilir ** </li><li> Reaksiyon süresi tamamlandıktan sonra, tüpler santrifüjlendi (5 dakika boyunca 12.000 x g) ve iki yıkama işlemleri uygulamaya robotik istasyonundan geri gönderilir. </li><li> Ikinci bir şırınga pompası şırınga soğuk su ile doldurulur ise yıkama için, bir şırınga boş ve ilk programlanabilir şırınga pompası yerleştirilmiş kalır. Her bir tüp içinde süpernatan boş bir şırınga pompası ve ikinci yataklarının soğuk su içine geri çekilir. </li><li> Nano HomojenizasyonAdım 2,6 tarif edildiği gibi parçacık süspansiyonu yapılır. Tüpler daha sonra santrifüj edilir (5 dakika boyunca 12.000 x g) ve 2,9 adım tarif edildiği gibi ikinci bir yıkama adımı gerçekleştirilir. </li><li> İkinci reaksiyon için, iki biriktirme adımları kullanılır. Birinci yerleştirme adımda, 12 eq. EDC bir pompa ve 12 eq ile yüklenir. NHS ikinci pompa ile yüklenir. </li><li> İkinci yerleştirme basamağı 10 eq içerir. Birinci ve ikinci pompaları (yani, galaktoz, laktoz ya da di-mannoz) ve *** 10 eq ile üçüncü bir pompa, bir spesifik bir sakarit. ve glikolik asit (kontrol olarak kullanılmıştır ****). </li><li> 4 ° C&#39;de adım 2.6 &#39;da açıklanan ve sürekli rotasyon ile 9 saat inkübe olarak Nanopartikül süspansiyonlar homojenize edilir </li><li> Reaksiyon süresi tamamlandıktan sonra adım 2.8, 2.9 ve 2.10 &#39;da anlatıldığı gibi, bir yıkama adımı gerçekleştirilir. </li><li> Fonksiyonlandırılmış nanoparçacık kütüphanesi sonra, en azından 2 saat boyunca kurumaya bir vakum hücresinin yerleştirilir. </li><li> Fonksiyonlandırılmış nanopartiküller sonra karakter olanX-elektron Spektroskopisi ve sırasıyla yüzey bileşimi ve sakarit konsantrasyonunu belirlemek için yüksek bir verim fenol-sülfürik asit deneyine göre ized. Elektron mikroskobu ve dinamik ışık saçılımı Tarama tane boyutu, boyut dağılımı ve yüzey yükü belirlemek için kullanılır. </li></ol> Notlar: * Biriktirme hacmi her tüpte bulunan nanoparçacıkların kütlesi ile değişir. Birinci ve ikinci reaksiyonlar için ** Reaksiyon süresi, nihai konsantrasyonu sakarit ayarlamak için değiştirilebilir. *** Her sakkarit istenen gruba bağlı olarak test tüpleri içine yatırılır. Koruması giderilmiş sakaritlerin zaten nanoparçacık yüzeyine daha takılabilir kovalent olarak bağlanmış bu molekülün, beri **** karbonhidrat tutturulması için bu çalışmada kullanılan belli bir tepkime için, glikolik asit, bir bağlayıcının kontrol olarak kullanılmıştır. 3. Temsilcisi Sonuçlar FulŞekil 2 de gösterildiği ly korunan dimannoside otomasyon platformu kullanılarak sentezlenmiştir. Sentezli bir bileşim çözücü olarak kullanarak, bir CDCl3 VXR 400 MHz spektrometre 1 H NMR ile karakterize edilmiştir. NMR spektrum Şekil 3 &#39;de gösterilmiştir. Polianhidrid yüksek verimlilik nanoparçacık imalat ve işlevsel kullanmak burada açıklanan nanoparçacıklar, dimannose, laktoz ve galaktoz eki başarıyla 10, 11 yapılmıştır. Bu set up kullanarak, optimum reaksiyon koşulları (yani, reaksiyon sıcaklığı ve zaman) istenen nanoparçacık fonksiyonlandırmalar ve morfolojisi ulaşmak için belirlenmiştir. Reaksiyon 4 azından gerçekleştirildiği zaman ° C yerine, oda sıcaklığı arasında, nanoparçacık yığılmasında bir azalma ile SEM ile gözlenmiştir (veri gösterilmemiştir). Di-mannoz veya biriyle CPH nanopartiküller: Tablo 1 fonksiyonlandırılmış 50:50 CPTEG karakterizasyonu temsilcisi sonuçlarını gösterir4 de sentezlenen laktoz, ° C. Veri fonksiyonlandırmalar nedeniyle ortalama nanoparçacık çapı küçük bir artış gösterir. Olmayan fonksiyonlandırılmış nanopartiküller yaklaşık bir negatif zeta potansiyeline sahipti. -20 MV, fonksiyonlandırılmış parçacıklar nanoparçacık yüzeyinin başarılı fonksiyonlandırmalar gösteren, pozitif zeta potansiyel değeri gösterdi. Laktoz ve di-mannoz hem de nötr şeker olmakla birlikte, etilen gelen serbest amin gruplarının sakaritlerin bağlamak için kullanılan bağlayıcı pozitif zeta potansiyelinin sorumlu olabilir diamin. Reaksiyon zamanı nanopartiküllerin son morfolojisi ve elde şeker bağlanma derecesi hem de etkileyebilecek bir diğer değişkendir. Reaksiyon süresi ayarlayarak, nanopartiküller yüzeyine iliştirilmiş nihai şekeri konsantrasyonları Şekil 4A&#39;da gösterildiği gibi, kontrol edilebilir. Beklenildiği gibi, 50:50 CPTEG yüzeyinde dimannose konsantrasyonu: CPH nanoparçacıklar artmıştırReaksiyon süresi ve toplam 18 saat sonra maksimum ulaşmıştır. 24 saat toplam reaksiyon zamanı ile fonksiyonelleştirilmiş Nanopartiküller fare kemik iliği kökenli dendritik hücreler (DC) üzerine CLRs hedef yeteneklerini değerlendirmek için kullanıldı. Flow sitometri iki CL reseptörleri (yani, Cire (CD209, DC-SIGN) ve mannoz reseptörü (CD206)) non-fonksiyonlandırılmış ile uyarılması sonrasında ve laktoz ve di-mannoz fonksiyonlandırılmış nanopartiküller (Şekil 4B) ifadesi değerlendirmek için kullanıldı. Hücreler laktoz ve di-mannoz fonksiyonlandırılmış nanopartiküller her ikisi ile uyarılan edildiğinde etkili hedefleme bir göstergesidir Her iki reseptör daha yüksek bir ifadesi, elde edildi. Bununla birlikte, di-mannoz-fonksiyonlu parçacıklar incelenmiştir reseptörleri için, bu bir ligant spesifite gösteren ifade daha yüksek bir seviyeye gösterdi. <table border="1"><tbody><tr><td> Nanopartikül tipi </td><td> Ortalama Partikül Çapı (nm) </td><td> Aveöfke Parçacık ζ-Potansiyeli (mV) </td></tr><tr><td> Sigara fonksiyonlandırılmış </td><td> 162 ± 43 </td><td> -20 ± 0.6 </td></tr><tr><td> Laktoz </td><td> 235 ± 34 </td><td> 26 ± 2.4 </td></tr><tr><td> Di-mannoz </td><td> 243 ± 32 </td><td> 30 ± 4.2 </td></tr></tbody></table> Tablo 1. Nanopartikül karakterizasyonu. Sigara fonksiyonlandırılmış ve fonksiyonlandırılmış yarı elastik ışık saçılması ve zeta potansiyel ölçümleri ile karakterize edildi. Parçacık boyutu verileri ortalama değer ± üç bağımsız deneyde toplanan dinamik ışık saçılımı verilerinin standart sapma (SD) temsil eder. Zeta potansiyel veriler ortalama değer ± SD üç bağımsız okuma temsil eder. CPH nanoparçacık yüzey: zeta potansiyelin işareti olarak değiştirin ki şeker 50:50 CPTEG verimli konjuge göstermektedir. <img alt="Şekil 1" src<html> = &quot;/ Files/ftp_upload/3967/3967fig1.jpg&quot; /&gt; Şekil 1. Karbonhidrat polianhidrid nanopartiküllerin fonksiyonlandırmalar ve açıklanan yüksek verimlilik yaklaşımı ile tasarlanmış olabilir fonksiyonlandırılmış nanoparçacık kütüphanelerin bir örnek ile izlenen yaklaşımın (A) grafiksel gösterimi. (B) otomatik yerleştirme tertibatı şematik gösterimi (i) Üç KD 1000 pompaları oluşan parçacık fonksiyonlandırmalar için kullanılan, (ii) bir robot aşamada iki aktüatörler (Zaber) tarafından entegre: x yönünde hareket için bir ve diğer iki komşu raf (tüpler ve küvetler için uygun) üç aktüatörler, her yön için bir (x, y ve z) oluşan ile (iii) bir ikinci aşama robotik; y yönde hareket için. Pompalar ve beş aktüatörler toplam seri olarak bağlanmış. Aktüatörler ve pompalar LabVIEW yazılımı kullanarak bir bilgisayar tarafından işletilmektedir. Bu şema ölçek değildir.arge.jpg &quot;target =&quot; _blank &quot;&gt; büyük rakamı görmek için buraya tıklayın. <img alt="Şekil 2" src="/files/ftp_upload/3967/3967fig2.jpg" /> Şekil 2. Otomatik grafiksel gösterimi örnek olarak mannoz kullanarak karbonhidrat sentezi iteratif. <img alt="Şekil 3" src="/files/ftp_upload/3967/3967fig3.jpg" /> Şekil 3. Korunan dimannoside 1 H-NMR. <img alt="Şekil 4" src="/files/ftp_upload/3967/3967fig4.jpg" /> Şekil 4. (A) Sakkarit ve nanoparçacık yüzey konsantrasyonu reaksiyon süresi etkisi. Gösterilen veriler, 50:50 CPTEG: CPH nanopartiküller farklı reaksiyon bazen dimannose ile fonksiyonelleştirilmiş edildi ve reaksiyon 4 gerçekleştirildi ° C. İki bağımsız fonksiyonlandırmalar deneylerin ortalama ve standart hata gösterilir. (B) Laktoz ve di-mannoz fonksiyonlandırılmış nanopartiküllerelde ifadesi ile karşılaştırıldığında CPH nanopartiküller: Kemik fonksiyonlandırılmış 50:50 CPTEG ile uyarıldıktan sonra bu iki belirteçlerin gelişmiş ifadesi gösterdiği gibi iliği kökenli dendritik hücreler üzerinde etkili hedef DC-SIGN (Cire, CD209) ve mannoz reseptörü (CD206) non-fonksiyonlu partikülleri ile.

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A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+dendritic+cells+with+biomaterials:+developing+the+next+generation+of+vaccines.">Targeting dendritic cells with biomaterials: developing the next generation of vaccines.</a> Trends Immunol. 27, 573-580 (2006).</li><li>Higashi, N., Fujioka, K., Denda-Nagai, K., Hashimoto, S., Nagai, S., Sato, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+macrophage+C-type+lectin+specific+for+galactose/N-acetylgalactosamine+is+an+endocytic+receptor+expressed+on+monocyte-derived+immature+dendritic+cells.">The macrophage C-type lectin specific for galactose/N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells.</a> J. Biol. Chem. 277, 20686 (2002).</li><li>Geijtenbeek, T. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Signalling+through+C-type+lectin+receptors:+shaping+immune+responses.">Signalling through C-type lectin receptors: shaping immune responses.</a> Nat. Rev. Immunol. 9, 465-479 (2009).</li><li>Seeberger, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+oligosaccharide+synthesis.">Automated oligosaccharide synthesis.</a> Chem. Soc. Rev. 37, 19-28 (2008).</li><li>Seeberger, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+Carbohydrate+Synthesis+as+Platform+to+Address+Fundamental+Aspects+of+Glycobiology-Current+Status+and+Future+Challenges.">Automated Carbohydrate Synthesis as Platform to Address Fundamental Aspects of Glycobiology-Current Status and Future Challenges.</a> Carb. Res. 343, 1889-1896 (2008).</li><li>Jaipuri, F. A., Pohl, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+solution-phase+automated+iterative+synthesis:+fluorous-tag+assisted+solution-phase+synthesis+of+linear+and+branched+mannose+oligomers.">Toward solution-phase automated iterative synthesis: fluorous-tag assisted solution-phase synthesis of linear and branched mannose oligomers.</a> Org. Biomol. Chem. 6, 2686-2691 (2008).</li><li>Petersen, L. K., Chavez-Santoscoy, A., 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+synthesis+of+and+high-throughput+protein+release+from+polymer+film+and+nanoparticle+libraries.">Combinatorial synthesis of and high-throughput protein release from polymer film and nanoparticle libraries.</a> J. Vis. Exp. , (2011).</li><li>Song, E. -. H., Osanya, A. O., Petersen, C. A., Pohl, N. L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+multivalent+tuberculosis+and+Leishmania-associated+capping+carbohydrates+reveals+structure-dependent+responses+allowing+immune+evasion.">Synthesis of multivalent tuberculosis and Leishmania-associated capping carbohydrates reveals structure-dependent responses allowing immune evasion.</a> J. Am. Chem. Soc. 132, 11428-11430 (2010).</li><li>Hakamori, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Aberrant+glycosylation+in+tumor+and+tumor+associated+carbohydrate+antigens.">Aberrant glycosylation in tumor and tumor associated carbohydrate antigens.</a> Adv. Cancer Res. 59, 257-331 (1989).</li><li>Atherton, T., Sheppard, R. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Solid-phase peptide synthesis: a practical approach. , (1999).</li><li>Caruthers, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gene+synthesis+machines:+DNA+chemistry+and+the+uses.">Gene synthesis machines: DNA chemistry and the uses.</a> Science. 230, 281-285 (1985).</li><li>Plante, O. J., Palmacci, E. R., Seeberger, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+solid-+phase+synthesis+of+oligosaccharides.">Automated solid- phase synthesis of oligosaccharides.</a> Science. 291, 1523-1527 (2001).</li><li>Ko, K. -. S., Park, G., Yu, Y., Pohl, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protecting+group-based+colorimetric+monitoring+of+fluorous-phase+and+solid-phase+synthesis+of+oligoglucosamines.">Protecting group-based colorimetric monitoring of fluorous-phase and solid-phase synthesis of oligoglucosamines.</a> Org. Lett. 10, 5381-5384 (2008).</li><li>Pohl, N. L., Chen, X. H. R., Wang, G. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Automated+solution-phase+oligosaccharide+synthesis+and+carbohydrate+microarrays:+development+of+fluorous-based+tools+for+glycomics.">Automated solution-phase oligosaccharide synthesis and carbohydrate microarrays: development of fluorous-based tools for glycomics.</a> Chemical Glycobiology. , 272-287 (2008).</li></ol>

NLBP kurucularından ve karbonhidrat şirket LuCella Biosciences, Inc özkaynak tutar

Bağışıklık hücreleri için doğrudan nanoparçacık etkileşimler maddeleri hedeflemek olarak karbonhidrat etkinliği, daha önce 10, 11 gösterilmiştir. Laboratuvarlarımızda Önceki araştırma ve böylece daha fazla T hücre aktivasyonunun 10, 11 için önemli olabilir bağışıklık hücreleri aktivasyonu arttırılması, polianhidrid nanopartiküller bağlı belirli şekerler antijen sunan hücreler (APC) üzerinde farklı CLRs hedeflemek için mümkün olduğunu göstermiştir. Ancak...

<table border="1"><tbody><tr><td> Isim </td><td> Şirket </td><td> Katalog numarası </td></tr><tr><td> Motorlu XYZ Aşama: 3x T-LSM050A, eksen başına 50 mm seyahat </td><td> Zaber Teknolojileri </td><td> T-XYZ-LSM050A-KT04 </td></tr><tr><td> KD-1000 Tek Şırınga Pompası </td><td> New Era Pompa Sistemleri </td><td> KD-1000 </td></tr><tr><td> Pyrex * Vista * Rimless Yeniden Cam Kültür Tüpler </td><td> Corning </td><td> 07-250-125 </td></tr><tr><td> ASW 1000 </td><td> Chemspeed Teknolojileri </td><td></td></tr><tr><td> LabVIEW </td><td> National Instruments </td><td> 776671-35 </td></tr><tr><td> SGE Gaz Geçirmez Şırınga, 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 tüp rotator </td><td> Fisher Scientific </td><td> 05-450-127 </td></tr></tbody></table>

high-throughput synthesis of carbohydrates and functionalization of polyanhydride nanoparticles

Bu malzeme tasarım, nanoteknoloji, kimya ve immünoloji gibi alanlarda ilgili Transdisipliner yaklaşımlar rasyonel etkili aşılar taşıyıcıları tasarlamak için kullanılmıştır gerekir. Nanopartikül-tabanlı platformlar aşı immünojenitesi 1 artırabilirsiniz aşı antijenleri sebat uzatabilir. Çeşitli biyo-bozunabilir polimerler aşı dağıtım araçları 1 olarak ele alınmıştır, özellikle de, polianhidrid partikülleri stabil protein antijenleri arasında sürekli bir şekilde tahliye edilmesini sağlayacak ve antijen sunan hücreler aktif hale getirmek ve bağışıklık yanıtı 2-12 modüle etmek için yeteneğini göstermişlerdir. Bu aşı taşıyıcıların moleküler tasarım polimerin özelliklerinin rasyonel seçimi gibi uygun bir hedef ajanların eklenmesi entegre gerekiyor. Ligandlar ve fonksiyonlandırılmış parçacıklar hedefliyorsunuz Yüksek throughput otomatik imalat geniş bir r çalışma yeteneğini artıracak güçlü bir araçtırözelliklerinin ange ve tekrarlanabilir bir aşı uygulama araçları tasarımına yol açacaktır. Bağışıklık hücreleri belirli reseptörleri tarafından kabul olma yeteneğine sahip ligandların hedefleme Ayrıca 10,11,13 C-tipi lektin reseptörleri (CLRs) üzerine karbonhidrat mevcut tanıması örüntü tanıma reseptörleri (PRR) olan modüle ve kişiye bağışıklık yanıtlarını gösterilmiştir patojenlerin yüzey. CLRs ile bağışıklık hücreleri antijene uyarılması ve daha da T hücresi aktivasyonu için 14,15 sonraki sunum geliştirilmiş interne sağlar. Bu nedenle, karbonhidrat molekülleri immün yanıtın çalışmada önemli bir rol oynar, ancak bu biyomoleküllerin kullanımı sıklıkla yapısal olarak iyi tanımlanmış ve saf karbonhidrat kullanılabilirlik eksikliğinden muzdarip. Çözüm fazlı reaksiyonlar iteratif dayalı bir otomasyon platformu anlamlı derecede düşük b kullanarak bu zorlu sentetik moleküllerin hızlı ve kontrollü sentezi etkinleştirebilirsinizGeleneksel bir katı-faz yöntemleri 16,17 &#39;den blok miktarlarda uilding. Bu yazıda böyle bir ara arıtma için fluorous katı-faz ekstraksiyon ile mannoz tabanlı ligandları olarak oligosakkarid otomatikleştirilmiş bir çözüm-faz sentezi için bir protokol rapor. Karbonhidrat bazlı hedeflemeyi ajan yapmak için otomatik yöntemler geliştirilmesi sonra, daha önce açıklandığı gibi 10 LabVIEW tarafından işletilen bir otomatik robotlu kurulum istihdam polianhidrid Nano partikülleri yüzeyinde bağlılığını yöntemleri açıklanmaktadır. Karbonhidrat ile yüzeyi fonksiyonlandırmalar CLRs 10,11 hedefleme ve ortaya çıkarmak bir çoklu-parametrik sistemi ile ilişkili karmaşıklıklara büyük bir değer (Şekil 1a) olacak kadar imalat yönteminin verim artan etkinlik göstermiştir.

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

Bu makale, yüksek bir verim yöntemi antijen sunan hücreler üzerinde belirli reseptörlerine yönelik olarak kullanım için polianhidrid Nanopartiküllerin yüzeyine oligosaccharidler ve ekin sentezi için sunulmuştur.

Polianhidrid Nanopartiküller Yüksek throughput Karbonhidrat sentezi ve işlevsel

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

high-throughput-synthesis-carbohydrates-functionalization

Research

JoVE Journal

Bioengineering

11.3K Görüntüleme.  Iowa State University. Bu makale, yüksek bir verim yöntemi antijen sunan hücreler üzerinde belirli reseptörlerine yönelik olarak kullanım için polianhidrid Nanopartiküllerin yüzeyine oligosaccharidler ve ekin sentezi için sunulmuştur.

Video: High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

Polymer Microarrays for High Throughput Discovery of Biomaterials

The discovery of novel biomaterials that are optimized for a specific biological application is readily achieved using polymer microarrays, which allows a combinatorial library of materials to be screened in a parallel, high throughput format1. Herein is described the formation and characterization of a polymer microarray using an on-chip photopolymerization technique 2. This involves mixing monomers at varied ratios to produce a library of monomer solutions, transferring the solution to a glass slide format using a robotic printing device and curing with UV irradiation. This format is readily amenable to many biological assays, including stem cell attachment and proliferation, cell sorting and low bacterial adhesion, allowing the ready identification of 'hit' materials that fulfill a specific biological criterion3-5. Furthermore, the use of high throughput surface characterization (HTSC) allows the biological performance to be correlated with physio-chemical properties, hence elucidating the biological-material interaction6. HTSC makes use of water contact angle (WCA) measurements, atomic force microscopy (AFM), X-ray photoelectron spectroscopy (XPS) and time-of-flight secondary ion mass spectrometry (ToF-SIMS). In particular, ToF-SIMS provides a chemically rich analysis of the sample that can be used to correlate the cell response with a molecular moiety. In some cases, the biological performance can be predicted from the ToF-SIMS spectra, demonstrating the chemical dependence of a biological-material interaction, and informing the development of hit materials5,3.

A description of the formation of a polymer microarray using an on-chip photopolymerization technique. The high throughput surface characterization using atomic force microscopy, water contact angle measurements, X-ray photoelectron spectroscopy and time of flight secondary ion mass spectrometry and a cell attachment assay is also described.

Biyomalzemelerin Yüksek Verimli Discovery için Polimer Mikroarray&#39;ler

The discovery of novel biomaterials that are optimized for a specific biological application is readily achieved using polymer microarrays, which allows a ...

Biyomühendislik

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

Polyanhydrides are a class of biomaterials with excellent biocompatibility and drug delivery capabilities. While they have been studied extensively with conventional one-sample-at-a-time synthesis techniques, a more recent high-throughput approach has been developed enabling the synthesis and testing of large libraries of polyanhydrides1. This will facilitate more efficient optimization and design process of these biomaterials for drug and vaccine delivery applications. The method in this work describes the combinatorial synthesis of biodegradable polyanhydride film and nanoparticle libraries and the high-throughput detection of protein release from these libraries. In this robotically operated method (Figure 1), linear actuators and syringe pumps are controlled by LabVIEW, which enables a hands-free automated protocol, eliminating user error. Furthermore, this method enables the rapid fabrication of micro-scale polymer libraries, reducing the batch size while resulting in the creation of multivariant polymer systems. This combinatorial approach to polymer synthesis facilitates the synthesis of up to 15 different polymers in an equivalent amount of time it would take to synthesize one polymer conventionally. In addition, the combinatorial polymer library can be fabricated into blank or protein-loaded geometries including films or nanoparticles upon dissolution of the polymer library in a solvent and precipitation into a non-solvent (for nanoparticles) or by vacuum drying (for films). Upon loading a fluorochrome-conjugated protein into the polymer libraries, protein release kinetics can be assessed at high-throughput using a fluorescence-based detection method (Figures 2 and 3) as described previously1. This combinatorial platform has been validated with conventional methods2 and the polyanhydride film and nanoparticle libraries have been characterized with 1H NMR and FTIR. The libraries have been screened for protein release kinetics, stability and antigenicity; in vitro cellular toxicity, cytokine production, surface marker expression, adhesion, proliferation and differentiation; and in vivo biodistribution and mucoadhesion1-11. The combinatorial method developed herein enables high-throughput polymer synthesis and fabrication of protein-loaded nanoparticle and film libraries, which can, in turn, be screened in vitro and in vivo for optimization of biomaterial performance.

This method describes the combinatorial synthesis of biodegradable polyanhydride film and nanoparticle libraries and the high-throughput detection of protein release from these libraries.

Kombinatorik Sentezi ve Polimer Film ve Nanoparçacık Kütüphaneler gelen Yüksek throughput Protein Yayın

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

Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles

Biodegradable nanoparticles have emerged as a versatile platform for the design and implementation of new intranasal vaccines against respiratory infectious diseases. Specifically, polyanhydride nanoparticles composed of the aliphatic sebacic acid (SA), the aromatic 1,6-bis(p-carboxyphenoxy)hexane (CPH), or the amphiphilic 1,8-bis(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) display unique bulk and surface erosion kinetics1,2 and can be exploited to slowly release functional biomolecules (e.g., protein antigens, immunoglobulins, etc.) in vivo3,4,5. These nanoparticles also possess intrinsic adjuvant activity, making them an excellent choice for a vaccine delivery platform6,7,8.
	In order to elucidate the mechanisms governing the activation of innate immunity following intranasal mucosal vaccination, one must evaluate the molecular and cellular responses of the antigen presenting cells (APCs) responsible for initiating immune responses. Dendritic cells are the principal APCs found in conducting airways, while alveolar macrophages (AM&#632;) predominate in the lung parenchyma9,10,11. AM&#632; are highly efficient in clearing the lungs of microbial pathogens and cell debris12,13. In addition, this cell type plays a valuable role in the transport of microbial antigens to the draining lymph nodes, which is an important first step in the initiation of an adaptive immune response9. AM&#632; also express elevated levels of innate pattern recognition and scavenger receptors, secrete pro-inflammatory mediators, and prime na&iuml;ve T cells12,14. A relatively pure population of AM&#632; (e.g., greater than 80%) can easily be obtained via lung lavage for study in the laboratory. Resident AM&#632; harvested from immune competent animals provide a representative phenotype of the macrophages that will encounter the particle-based vaccine in vivo. Herein, we describe the protocols used to harvest and culture AM&#632; from mice and examine the activation phenotype of the macrophages following treatment with polyanhydride nanoparticles in vitro.

Herein, we describe protocols for harvesting murine alveolar macrophages, which are resident innate immune cells in the lung, and examining their activation in response to co-culture with polyanhydride nanoparticles.

Polianhidrid Nanopartiküller Bağlı Mürin Alveoler Makrofajlar Hasat ve Değerlendirme Hücresel Etkinleştirme

Biodegradable nanoparticles have emerged as a  versatile platform for the design and implementation of new intranasal vaccines  against respiratory ...

High Throughput Single-cell and Multiple-cell Micro-encapsulation

Microfluidic encapsulation methods have been previously utilized to capture cells in picoliter-scale aqueous, monodisperse drops, providing confinement from a bulk fluid environment with applications in high throughput screening, cytometry, and mass spectrometry. We describe a method to not only encapsulate single cells, but to repeatedly capture a set number of cells (here we demonstrate one- and two-cell encapsulation) to study both isolation and the interactions between cells in groups of controlled sizes. By combining drop generation techniques with cell and particle ordering, we demonstrate controlled encapsulation of cell-sized particles for efficient, continuous encapsulation. Using an aqueous particle suspension and immiscible fluorocarbon oil, we generate aqueous drops in oil with a flow focusing nozzle. The aqueous flow rate is sufficiently high to create ordering of particles which reach the nozzle at integer multiple frequencies of the drop generation frequency, encapsulating a controlled number of cells in each drop. For representative results, 9.9 &mu;m polystyrene particles are used as cell surrogates. This study shows a single-particle encapsulation efficiency Pk=1 of 83.7% and a double-particle encapsulation efficiency Pk=2 of 79.5% as compared to their respective Poisson efficiencies of 39.3% and 33.3%, respectively. The effect of consistent cell and particle concentration is demonstrated to be of major importance for efficient encapsulation, and dripping to jetting transitions are also addressed. 
Introduction 
Continuous media aqueous cell suspensions share a common fluid environment which allows cells to interact in parallel and also homogenizes the effects of specific cells in measurements from the media. High-throughput encapsulation of cells into picoliter-scale drops confines the samples to protect drops from cross-contamination, enable a measure of cellular diversity within samples, prevent dilution of reagents and expressed biomarkers, and amplify signals from bioreactor products. Drops also provide the ability to re-merge drops into larger aqueous samples or with other drops for intercellular signaling studies.1,2 The reduction in dilution implies stronger detection signals for higher accuracy measurements as well as the ability to reduce potentially costly sample and reagent volumes.3 Encapsulation of cells in drops has been utilized to improve detection of protein expression,4 antibodies,5,6 enzymes,7 and metabolic activity8 for high throughput screening, and could be used to improve high throughput cytometry.9 Additional studies present applications in bio-electrospraying of cell containing drops for mass spectrometry10 and targeted surface cell coatings.11 Some applications, however, have been limited by the lack of ability to control the number of cells encapsulated in drops. Here we present a method of ordered encapsulation12 which increases the demonstrated encapsulation efficiencies for one and two cells and may be extrapolated for encapsulation of a larger number of cells.
To achieve monodisperse drop generation, microfluidic "flow focusing" enables the creation of controllable-size drops of one fluid (an aqueous cell mixture) within another (a continuous oil phase) by using a nozzle at which the streams converge.13 For a given nozzle geometry, the drop generation frequency f and drop size can be altered by adjusting oil and aqueous flow rates Qoil and Qaq. As the flow rates increase, the flows may transition from drop generation to unstable jetting of aqueous fluid from the nozzle.14 
When the aqueous solution contains suspended particles, particles become encapsulated and isolated from one another at the nozzle. For drop generation using a randomly distributed aqueous cell suspension, the average fraction of drops Dk containing k cells is dictated by Poisson statistics, where Dk = &lambda;k exp(-&lambda;)/(k!) and &lambda; is the average number of cells per drop. The fraction of cells which end up in the "correctly" encapsulated drops is calculated using Pk = (k x Dk)/&Sigma;(k' x Dk'). The subtle difference between the two metrics is that Dk relates to the utilization of aqueous fluid and the amount of drop sorting that must be completed following encapsulation, and Pk relates to the utilization of the cell sample. As an example, one could use a dilute cell suspension (low &lambda;) to encapsulate drops where most drops containing cells would contain just one cell. While the efficiency metric Pk would be high, the majority of drops would be empty (low Dk), thus requiring a sorting mechanism to remove empty drops, also reducing throughput.15
Combining drop generation with inertial ordering provides the ability to encapsulate drops with more predictable numbers of cells per drop and higher throughputs than random encapsulation. Inertial focusing was first discovered by Segre and Silberberg16 and refers to the tendency of finite-sized particles to migrate to lateral equilibrium positions in channel flow. Inertial ordering refers to the tendency of the particles and cells to passively organize into equally spaced, staggered, constant velocity trains. Both focusing and ordering require sufficiently high flow rates (high Reynolds number) and particle sizes (high Particle Reynolds number).17,18 Here, the Reynolds number Re =uDh/&nu; and particle Reynolds number Rep =Re(a/Dh)2, where u is a characteristic flow velocity, Dh [=2wh/(w+h)] is the hydraulic diameter, &nu; is the kinematic viscosity, a is the particle diameter, w is the channel width, and h is the channel height. Empirically, the length required to achieve fully ordered trains decreases as Re and Rep increase. Note that the high Re and Rep requirements (for this study on the order of 5 and 0.5, respectively) may conflict with the need to keep aqueous flow rates low to avoid jetting at the drop generation nozzle. Additionally, high flow rates lead to higher shear stresses on cells, which are not addressed in this protocol. The previous ordered encapsulation study demonstrated that over 90% of singly encapsulated HL60 cells under similar flow conditions to those in this study maintained cell membrane integrity.12 However, the effect of the magnitude and time scales of shear stresses will need to be carefully considered when extrapolating to different cell types and flow parameters. The overlapping of the cell ordering, drop generation, and cell viability aqueous flow rate constraints provides an ideal operational regime for controlled encapsulation of single and multiple cells.
Because very few studies address inter-particle train spacing,19,20 determining the spacing is most easily done empirically and will depend on channel geometry, flow rate, particle size, and particle concentration. Nonetheless, the equal lateral spacing between trains implies that cells arrive at predictable, consistent time intervals. When drop generation occurs at the same rate at which ordered cells arrive at the nozzle, the cells become encapsulated within the drop in a controlled manner. This technique has been utilized to encapsulate single cells with throughputs on the order of 15 kHz,12 a significant improvement over previous studies reporting encapsulation rates on the order of 60-160 Hz.4,15 In the controlled encapsulation work, over 80% of drops contained one and only one cell, a significant efficiency improvement over Poisson (random) statistics, which predicts less than 40% efficiency on average.12
In previous controlled encapsulation work,12 the average number of particles per drop &lambda; was tuned to provide single-cell encapsulation. We hypothesize that through tuning of flow rates, we can efficiently encapsulate any number of cells per drop when &lambda; is equal or close to the number of desired cells per drop. While single-cell encapsulation is valuable in determining individual cell responses from stimuli, multiple-cell encapsulation provides information relating to the interaction of controlled numbers and types of cells. Here we present a protocol, representative results using polystyrene microspheres, and discussion for controlled encapsulation of multiple cells using a passive inertial ordering channel and drop generation nozzle.

Combining monodisperse drop generation with inertial ordering of cells and particles, we describe a method to encapsulate a desired number of cells or particles in a single drop at kHz rates. We demonstrate efficiencies twice exceeding those of unordered encapsulation for single- and double-particle drops.

Yüksek Verim Micro-encapsulation Tek hücreli ve Çoklu-hücre

Microfluidic encapsulation methods have been previously utilized to capture cells in picoliter-scale aqueous, monodisperse drops, providing confinement ...

Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry

Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases1 and as a technology for regenerative medicine2. Unlike viruses, which have significant safety issues, polymeric nanoparticles can be designed to be non-toxic, non-immunogenic, non-mutagenic, easier to synthesize, chemically versatile, capable of carrying larger nucleic acid cargo and biodegradable and/or environmentally responsive. Cationic polymers self-assemble with negatively charged DNA via electrostatic interaction to form complexes on the order of 100 nm that are commonly termed polymeric nanoparticles. Examples of biomaterials used to form nanoscale polycationic gene delivery nanoparticles include polylysine, polyphosphoesters, poly(amidoamines)s and polyethylenimine (PEI), which is a non-degradable off-the-shelf cationic polymer commonly used for nucleic acid delivery1,3 . Poly(beta-amino ester)s (PBAEs) are a newer class of cationic polymers4 that are hydrolytically degradable5,6 and have been shown to be effective at gene delivery to hard-to-transfect cell types such as human retinal endothelial cells (HRECs)7, mouse mammary epithelial cells8, human brain cancer cells9 and macrovascular (human umbilical vein, HUVECs) endothelial cells10.
A new protocol to characterize polymeric nanoparticles utilizing nanoparticle tracking analysis (NTA) is described. In this approach, both the particle size distribution and the distribution of the number of plasmids per particle are obtained11. In addition, a high-throughput 96-well plate transfection assay for rapid screening of the transfection efficacy of polymeric nanoparticles is presented. In this protocol, poly(beta-amino ester)s (PBAEs) are used as model polymers and human retinal endothelial cells (HRECs) are used as model human cells. This protocol can be easily adapted to evaluate any polymeric nanoparticle and any cell type of interest in a multi-well plate format.

A protocol for nanoparticle tracking analysis (NTA) and high-throughput flow cytometry to evaluate polymeric gene delivery nanoparticles is described. NTA is utilized to characterize the nanoparticle particle size distribution and the plasmid per particle distribution. High-throughput flow cytometry enables quantitative transfection efficacy evaluation for a library of gene delivery biomaterials.

Polimerik Gen Taşıyıcı Değerlendirilmesi Nanoparçacık İzleme Analizi ve Yüksek throughput Flow Sitometri tarafından Nanopartiküller

Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases1 and as a ...

High Throughput Microfluidic Rapid and Low Cost Prototyping Packaging Methods

In this work, 3 different packaging and assembly techniques are presented. They can be classified into two categories: one-time use and reusable packaging techniques.
The one-time use packaging technique employs UV-based and temperature curing epoxies to connect microtubes to access holes, wire-bonding for integrated circuit connections, and silver epoxy for electrical connections. This method is based on a robust assembly technique that can support relatively high pressure close to 1 psi and does not need any support to strengthen the microfluidic architecture.
Reusable packaging techniques consist of PDMS-based microtube interconnectors and anisotropic adhesive films for electrical connections. These devices are more sensitive and fragile. Consequently, Plexiglas support is added to the microfluidic structure to improve the electrical contact when anisotropic adhesive films are used, and also to strengthen the microfluidic architecture. In addition, a micromanipulator is needed to maintain tubes while using a thin PDMS layer to connect them to the access holes. Different PDMS layer thicknesses, ranging from 0.45-3 mm, are tested to compare the best adherence versus injection rates. Applied injection rates are varied from 50-300 &#956;l/hr for 0.45-3 mm PDMS layers, respectively. These techniques are mainly applicable for low-pressure applications. However, they can be extended for high-pressure ones through plasma-oxygen process to permanently seal the PDMS to glass substrates. The main advantage of this technique, besides the fact that it is reusable, consists of keeping the device observable when the microchannel length is very short (in the range of 3 mm or lower).

In this article we describe different techniques for microfluidic rapid prototyping platforms. The proposed techniques are based on ultraviolet (UV) sensitive and temperature curing epoxies, polydimethylsiloxane (PDMS) based tubing, wire-bonding, and anisotropic adhesive films. The assembling procedures presented are developed for both one-time use devices as well as reusable microfluidic systems.

In this work, 3 different packaging and assembly techniques are presented. They can be classified into two categories: one-time use and reusable packaging ...

Microwave-driven Synthesis of Iron Oxide Nanoparticles for Fast Detection of Atherosclerosis

A fast and reproducible microwave-driven protocol has been developed for the synthesis of neridronate-functionalized nanoparticles. Starting from the synthesis of hydrophobic nanoparticles, our method is based on an adaptation from thermal decomposition method to microwave driven synthesis. The new methodology produces a decrease in the reaction times in comparison with traditional procedures. Moreover, the use of the microwave technology increases the reproducibility of the reactions, something important from the point of view of clinical applications. The novelty of this iron oxide nanoparticle is the attachment of Neridronate. The use of this molecule leads a bisphosphonate moiety towards the outside of the nanoparticle that provides Ca2+ binding properties in vitro and selective accumulation in vivo in the atheroma plaque. The protocol allows the synthesis and plaque detection in about 3 hr since the initial synthesis from organic precursors. Their accumulation in the atherosclerotic area in less than 1 hr provides a contrast agent particularly suitable for clinical applications.

Microwave technology enables extremely fast synthesis of iron oxide nanoparticles for atherosclerosis plaque characterization. The use of an aminobisphosphonate in the external side of the nanoparticle provides a fast accumulation in the atherosclerotic area.

Ateroskleroz Hızlı Algılama Demir Oksit Nanopartiküller Mikrodalga odaklı sentezi

A fast and reproducible microwave-driven protocol has been developed for the synthesis of neridronate-functionalized nanoparticles. Starting from the ...

High-throughput Identification of Bacteria Repellent Polymers for Medical Devices

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as contributing to antimicrobial resistance. One possible avenue for the reduction of device-associated infections is the identification of bacteria-repellent polymer coatings for these devices, which would prevent bacterial binding at the initial attachment step. A method for the identification of such repellent polymers, based on the parallel screening of hundreds of polymers using a microarray, is described here. This high-throughput method resulted in the identification of a range of promising polymers that resisted binding of various clinically relevant bacterial species individually and also as multi-species communities. One polymer, PA13 (poly(methylmethacrylate-co-dimethylacrylamide)), demonstrated significant reduction in attachment of a number of hospital isolates when coated onto two commercially available central venous catheters. The method described could be applied to identify polymers for a wide range of applications in which modification of bacterial attachment is important.

A high-throughput microarray method for the identification of polymers which reduce bacterial surface binding on medical devices is described.

Tıbbi Cihazlar Bakterilerin yüksek verimlilik Kimlik Savar Polimerler

Medical devices are often associated with hospital-acquired infections, which place enormous strain on patients and the healthcare system as well as ...

Surface Functionalization of Hepatitis E Virus Nanoparticles Using Chemical Conjugation Methods

Virus-like particles (VLPs) have been used as nanocarriers to display foreign epitopes and/or deliver small molecules in the detection and treatment of various diseases. This application relies on genetic modification, self-assembly, and cysteine conjugation to fulfill the tumor-targeting application of recombinant VLPs. Compared with genetic modification alone, chemical conjugation of foreign peptides to VLPs offers a significant advantage because it allows a variety of entities, such as synthetic peptides or oligosaccharides, to be conjugated to the surface of VLPs in a modulated and flexible manner without alteration of the VLP assembly.
Here, we demonstrate how to use the hepatitis E virus nanoparticle (HEVNP), a modularized theranostic capsule, as a multifunctional delivery carrier. Functions of HEVNPs include tissue-targeting, imaging, and therapeutic delivery. Based on the well-established structural research of HEVNP, the structurally independent and surface-exposed residues were selected for cysteine replacement as conjugation sites for maleimide-linked chemical groups via thiol-selective linkages. One particular cysteine-modified HEVNP (a Cys replacement of the asparagine at 573 aa (HEVNP-573C)) was conjugated to a breast cancer cell-specific ligand, LXY30 and labeled with near-infrared (NIR) fluorescence dye (Cy5.5), rendering the tumor-targeted HEVNPs as effective diagnostic capsules (LXY30-HEVNP-Cy5.5). Similar engineering strategies can be employed with other macromolecular complexes with well-known atomic structures to explore potential applications in theranostic delivery.

We have engineered the capsid protein of hepatitis E virus as a theranostic nanoparticle (HEVNP). HEVNP self-assembles into a stable icosahedral cage in mucosal delivery. Here, we describe the modification of HEVNPs for tumor targeting by mutating surface-exposed residues to cysteines, which conjugate synthetic ligands that specifically bind tumor cells.

Hepatit E virüs nano tanecikleri kimyasal konjugasyon yöntemlerle yüzey Functionalization

Virus-like particles (VLPs) have been used as nanocarriers to display foreign epitopes and/or deliver small molecules in the detection and treatment of ...

Synthesis of Multi-walled Carbon Nanotubes Modified with Silver Nanoparticles and Evaluation of Their Antibacterial Activities and Cytotoxic Properties

In this study, multi-walled carbon nanotubes (MWCNTs) were treated with an aqueous sulfuric acid solution to form an oxygen-based functional group. Silver MWCNTs were prepared by the reductive deposition of silver from an aqueous solution of AgNO3 on the oxidized MWCNTs. Given the unique color of the CNTs, it was not possible to apply them to the minimum inhibitory concentration or mitochondrial toxicity assays to evaluate the toxicity and antibacterial properties, since they would interfere with the assays. The inhibition zone and minimum bactericidal concentration for the Ag-MWCNTs were measured and Live/Dead and Trypan Blue assays were used to measure the toxicity and antibacterial properties without interfering with the color of the CNTs.

In this study, antimicrobial nanomaterials were synthesized by acidic oxidation of multiwalled carbon nanotubes and subsequent reductive deposition of silver nanoparticles. Antimicrobial activity and cytotoxicity tests were performed with the as-prepared nanomaterials.

Gümüş nano tanecikleri ve onların antibakteriyel faaliyetleri ve sitotoksik özellikleri değerlendirilmesi ile modifiye çok duvarlı karbon nanotüpler sentezi

In this study, multi-walled carbon nanotubes (MWCNTs) were treated with an aqueous sulfuric acid solution to form an oxygen-based functional group. Silver ...