और उच्च throughput बहुलक फिल्म और Nanoparticle पुस्तकालय से प्रोटीन रिलीज की मिश्रित संश्लेषण
Summary
इस विधि biodegradable polyanhydride फिल्म और nanoparticle पुस्तकालयों की मिश्रित संश्लेषण और उच्च throughput इन पुस्तकालयों से प्रोटीन रिहाई का पता लगाने का वर्णन करता है.
Abstract
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.
Protocol
Discussion
ज्ञान आवश्यक संश्लेषण शर्तों और जा रहा संश्लेषित पॉलिमर के कांच संक्रमण तापमान (टी एंड जी) के पुस्तकालय के निर्माण के लिए आवश्यक हैं. यदि टी छ कमरे के तापमान से नीचे हैं, nanoparticle निर्माण कदम के लिए ?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
लेखकों ONR – मूरी (NN00014-06-1-१,१७६) और वित्तीय सहायता के लिए अमेरिकी सेना चिकित्सा अनुसंधान और materiel कमान (अनुदान सं W81XWH-10-1-0806) पुरस्कार को स्वीकार करते हैं. इस सामग्री अनुदान सं 0552584 और 0851519 EEC के तहत राष्ट्रीय विज्ञान फाउंडेशन द्वारा समर्थित काम पर आधारित 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 |
| Glass cuvettes | Scientific Strategies | G102 |
| LabVIEW | National Instruments | 776671-35 |
| SGE Gas Tight Syringes, Luer Loc | Sigma Aldrich | 509507 |
| U96 DeepWell Plates 1.3 ml & 2.0 ml | Thermo Scientific: Nunc | 278743 |
| Well cap mats | Thermo Scientific: Nunc | 276000 |
| Typhoon 9400 | GE Healthcare | 63-0055-79 |
| Whatman Grade 50 Circles 90 mm | Whatman | 1450-090 |
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