Vauthier Christine
Institut Galien Paris Sud, UMR CNRS 8612, University of Paris-Sud, Chatenay-Malabry, France.
Methods Mol Biol. 2019;1974:181-194. doi: 10.1007/978-1-4939-9220-1_13.
This chapter describes the preparation of chitosan-coated poly(isobutylcyanoacrylate) nanoparticles as a suitable carrier to deliver siRNAs to two types of xenograft tumor models of mice. The nanoparticles are prepared by a method of emulsion polymerization that includes steps of polymerization and purification. The polymerization method is carried out in a single pot in an aqueous medium. siRNAs are coupled with the nanoparticles at the end of the preparation by adsorption. The protocol also explains how to determine optimum yield/the titer of association of siRNA with the nanoparticles. It is described for a preparation scale at 4 mL of nanoparticle dispersion at a concentration of 42-46 mg nanoparticles/mL. Optimal loading capacity of the nanoparticles with the siRNA can be achieved by performing an association yield above 90% using a mass ratio of 1 mg siRNA/50 mg of nanoparticles (20 μg siRNA/mg nanoparticles, 1 nmol siRNA (M 14 kDa)/mg nanoparticles).
本章描述了壳聚糖包被的聚（异丁基氰基丙烯酸酯）纳米颗粒的制备方法，该纳米颗粒是一种合适的载体，可将小干扰RNA（siRNA）递送至两种小鼠异种移植肿瘤模型。纳米颗粒通过乳液聚合法制备，该方法包括聚合和纯化步骤。聚合方法在水性介质中的单锅中进行。在制备结束时，通过吸附将siRNA与纳米颗粒偶联。该方案还解释了如何确定siRNA与纳米颗粒结合的最佳产率/滴度。它描述了在4 mL纳米颗粒分散液（浓度为42 - 46 mg纳米颗粒/mL）的制备规模下的情况。通过使用1 mg siRNA/50 mg纳米颗粒的质量比（20 μg siRNA/mg纳米颗粒，1 nmol siRNA（分子量14 kDa）/mg纳米颗粒）进行结合产率高于90%的操作，可以实现纳米颗粒与siRNA的最佳负载能力。
