生产近红外敏感、核心壳疫苗输送平台
Summary
本文介绍了用于生产新型疫苗输送平台”多泡”的协议，以实现延迟突发释放。聚酯（乳酸共糖酸）和多丙酮用于形成多泡，小分子和抗原用作货物。
Abstract
疫苗交付战略可以限制货物接触有机溶剂，同时实现新的释放概况，对于提高全球免疫覆盖率至关重要。这里引进了一种新型的注射、紫外线固化和延迟突发释放-使疫苗输送平台，称为多泡。货物被注入聚酯基聚泡，形成于10%的碳基纤维素为基础的水溶液。本文包括保持多泡球形和优化货物放置和保留的协议，以最大限度地提高多泡内的货物量。为确保安全，使用中子活化分析对多泡中的氯化溶剂含量进行了分析。释放研究以小分子作为货物在多泡内进行，以确认延迟的爆裂释放。为了进一步展示货物按需交付的潜力，将金纳米棒混合在聚合物壳中，以实现近红外激光活化。
Introduction
免疫覆盖率有限导致300万人死亡，具体由疫苗可预防疾病引起1。储存和运输条件不足导致功能性疫苗的浪费，从而导致全球免疫接种减少。此外，由于未遵守要求的疫苗计划而不完全接种疫苗也会导致疫苗覆盖率有限，特别是在发展中国家。在接受强化注射的建议期间内，需要多次看望医务人员，从而限制完全接种疫苗的人口比例。因此，有必要为控制性疫苗的交付制定新的战略，以规避这些挑战。
目前开发疫苗交付技术的努力包括基于乳液的聚合物系统3，4。3,然而，货物往往暴露在更多的有机溶剂，可能会导致聚集和变性，特别是在蛋白质为基础的货物5，6,的背景下。我们开发了一种新型的疫苗输送平台”多泡”，可以潜在地装着多个货舱，同时最大限度地减少暴露于溶剂7中的货物数量。例如，在我们的多泡芯壳平台中，一个直径为 0.38 mm （SEM） 的货物口袋被注入 1 mm 多泡的中心。在这种情况下，暴露于有机溶剂的货物表面积约为0.453 mm2。在考虑球体（微粒）的包装密度后（货物库），可放入仓库的微粒（直径为10 μm）的实际体积为0.17 mm3。一个微粒的体积为5.24×10-8 mm3，因此可以容纳仓库的颗粒数量为+3.2×106颗粒。-8如果每个微粒有 20 个直径为 0 . 25 μm的货物口袋（由于液），则暴露于有机溶剂中的货物表面积为 1274 mm2。因此，与微粒中有机溶剂暴露的货物相比，多泡内的货库暴露于有机溶剂的表面面积要少2800倍。因此，我们的聚酯平台可能会减少暴露于有机溶剂的货物数量，否则会导致货物聚集和不稳定。
聚泡基于相分离原理形成，有机相中的聚酯被注入水溶液，形成球形气泡。水相中的货物可以注入多泡的中心。通过将不同的货物与聚合物壳混合，可以在多泡体中实现另一个货舱。在这个阶段，多泡将是可塑的，然后将被固化，导致一个坚实的多泡结构与货物在中间。选择球面多泡而不是其他几何形状，以增加多泡内的货物容量，同时最小化多泡的整体大小。选择在中心有货物的多泡来演示延迟爆裂释放。多泡体还与近红外（NIR）-敏感（即启用辐射）剂（即金纳米棒（AuNR）结合使用，导致多泡温度升高。这种效应可能促进更快的降解，并可用于控制未来应用中的动力学。在这篇论文中，我们描述了我们形成和描述多泡的方法，实现从多泡的延迟突发释放，并将AuNR纳入多泡中，导致NR激活。
Protocol
Representative Results
Discussion
当前技术和挑战
基于乳化的微颗粒和纳米粒子通常被用作药物输送载体。虽然从这些装置释放货物的动力学已经进行了广泛的研究，控制爆裂释放动力学一直是一个重大挑战11。由于货物接触过量的水性和有机溶剂，基于乳液的系统也限制了货物的多功能性和功能性。由于货物变性和聚合的可能性，基于蛋白质的货物通常与微粒子和纳米粒子不兼容</…
Divulgazioni
The authors have nothing to disclose.
Acknowledgements
我们要感谢B bryan E. Tomlin博士，他隶属于TAMU化学系的元素分析实验室，他协助了中子活化分析（NAA）。
Materials
| 1-Step Ultra Tetramethylbezidine (TMB)-Enzyme-Linked Immunosorbent Assay (ELISA) Substrate Solution | Thermo scientific | 34028 | |
| 2-Hydroxy-2-methylpropiophenone | TCI AMERICA | H0991 | |
| 450 nm Stop Solution for TMB Substrate | Abcam | ab17152 | |
| Acryloyl chloride | Sigma Aldrich | A24109-100G | |
| Acriflavine | Chem-Impex International | 22916 | |
| Anhydrous ethyl ether | Fisher Chemical | E138-500 | |
| Anti-HIV1 gp120 antibody conjugated to horseradish peroxidase (HRP) | |||
| Bovine serum albumin (BSA) | Fisher BioReagents | BP9700100 | |
| BSA-CF488 dye conjugates | Invitrogen | A13100 | |
| Bromosalicylic acid | Acros Organics | AC162142500 | |
| Carboxymethylcellulose (CMC) | Millipore Sigma | 80502-040 | |
| Centrimonium bromide (CTAB) | MP Biomedicals | ICN19400480 | |
| Chloroform | Fisher Chemical | C2984 | |
| Coating buffer | Abcam | ab210899 | |
| Dichloromethane (DCM) | Sigma Aldrich | 270997-1L | |
| Diethyl ether | Fisher Chemical | E1384 | |
| Dodeacyl Amine | Acros Organics | AC117665000 | |
| Doxorubicin hydrochloride | Fisher BioReagents | BP251610 | |
| L-ascorbic acid | Acros Organics | A61 100 | |
| Legato 100 Syringe Pump | KD Scientific | 14 831 212 | |
| mPEG thiol | Laysan Bio | NC0702454 | |
| Nonfat dry milk | Andwin Scientific | NC9022655 | |
| Potassium carbonate | Acros Organics | AC424081000 | |
| Phosphate saline buffer | Fisher BioReagents | BP3991 | |
| (Poly(caprolactone) | Sigma Aldrich | 440744-250G | |
| (Poly(caprolactone) triol | Acros Organics | AC190730250 | |
| Poly (lactic-co-glycolic acid) diacrylate | CMTec | 280050 | |
| Potassium carbonate | Acros Organics | AC424081000 | |
| Recombinant HIV1 gp120 + gp41 protein | Abcam | ab49054 | |
| Silver nitrate | Acros Organics | S181 25 | |
| Sodium borohydride | Fisher Chemical | S678 10 | |
| Tetrachloroauric acid | Fisher Chemical | G54 1 | |
| Trehalose | Acros Organics | NC9022655 | |
| Triethyl amine | Acros Organics | AC157910010 |
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