Adamo Giorgia, Grimaldi Natascia, Sabatino Maria Antonietta, Walo Marta, Dispenza Clelia, Ghersi Giulio
Biol Chem. 2017 Feb 1;398(2):277-287. doi: 10.1515/hsz-2016-0255.
Poly(N-vinyl pyrrolidone)-based-nanogels (NGs), produced by e-beam irradiation, are conjugated with monoclonal antibodies (mAb) for active targeting purposes. The uptake of immuno-functionalized nanogels is tested in an endothelial cell line, ECV304, using confocal and epifluorescence microscopy. Intracellular localization studies reveal a faster uptake of the immuno-nanogel conjugate with respect to the 'bare' nanogel. The specific internalization pathway of these immuno-nanogels is clarified by selective endocytosis inhibition experiments, flow cytometry and confocal microscopy. Active targeting ability is also verified by conjugating a monoclonal antibody which recognizes the αvβ3 integrin on activated endothelial cells. Epifluorescence images of the 'wound healing assay' on ECV304 cells provide evidence of nanogels localization only in the target cells. Therefore, the immuno-nanogels produced have the potential to recognize specific cell types in heterogeneous systems, which makes them promising candidates for targeted drug delivery applications.
通过电子束辐照制备的基于聚(N-乙烯基吡咯烷酮)的纳米凝胶(NGs)与单克隆抗体(mAb)偶联以用于主动靶向目的。使用共聚焦和落射荧光显微镜在内皮细胞系ECV304中测试免疫功能化纳米凝胶的摄取。细胞内定位研究表明,免疫纳米凝胶偶联物比“裸”纳米凝胶摄取更快。通过选择性内吞抑制实验、流式细胞术和共聚焦显微镜阐明了这些免疫纳米凝胶的特定内化途径。通过偶联识别活化内皮细胞上αvβ3整合素的单克隆抗体也验证了主动靶向能力。ECV304细胞上“伤口愈合试验”的落射荧光图像提供了纳米凝胶仅定位在靶细胞中的证据。因此,所制备的免疫纳米凝胶具有识别异质系统中特定细胞类型的潜力,这使其成为靶向药物递送应用的有前景的候选物。