Sánchez-Sánchez Lorena, Tapia-Moreno Alejandro, Juarez-Moreno Karla, Patterson Dustin P, Cadena-Nava Ruben D, Douglas Trevor, Vazquez-Duhalt Rafael
Instituto de Biotecnología, Universidad Nacional Autónoma de México, 62250, Cuernavaca, Morelos, Mexico.
Centro de Nanociencias y Nanotecnología, Universidad Nacional Autónoma de México, Km 107 carretera Tijuana-Ensenada, 22860, Ensenada, Baja California, Mexico.
J Nanobiotechnology. 2015 Oct 9;13:66. doi: 10.1186/s12951-015-0127-z.
BACKGROUND: The intracellular delivery of enzymes for therapeutic use has a promising future for the treatment of several diseases such as genetic disorders and cancer. Virus-like particles offer an interesting platform for enzymatic delivery to targeted cells because of their great cargo capacity and the enhancement of the biocatalyst stability towards several factors important in the practical application of these nanoparticles. RESULTS: We have designed a nano-bioreactor based on the encapsulation of a cytochrome P450 (CYP) inside the capsid derived from the bacteriophage P22. An enhanced peroxigenase, CYPBM3, was selected as a model enzyme because of its potential in enzyme prodrug therapy. A total of 109 enzymes per capsid were encapsulated with a 70 % retention of activity for cytochromes with the correct incorporation of the heme cofactor. Upon encapsulation, the stability of the enzyme towards protease degradation and acidic pH was increased. Cytochrome P450 activity was delivered into Human cervix carcinoma cells via transfecting P22-CYP nanoparticles with lipofectamine. CONCLUSION: This work provides a clear demonstration of the potential of biocatalytic virus-like particles as medical relevant enzymatic delivery vehicles for clinical applications.
背景:用于治疗用途的酶的细胞内递送对于治疗多种疾病(如遗传疾病和癌症)具有广阔的前景。病毒样颗粒因其巨大的载物能力以及生物催化剂对这些纳米颗粒实际应用中重要的多种因素的稳定性增强,为将酶递送至靶细胞提供了一个有趣的平台。 结果:我们设计了一种基于将细胞色素P450(CYP)封装在源自噬菌体P22的衣壳内的纳米生物反应器。由于其在酶前药疗法中的潜力,选择了一种增强型过氧化物酶CYPBM3作为模型酶。每个衣壳共封装了109个酶,对于正确掺入血红素辅因子的细胞色素,其活性保留率为70%。封装后,酶对蛋白酶降解和酸性pH的稳定性增加。通过用脂质体转染P22-CYP纳米颗粒,细胞色素P450活性被递送至人宫颈癌细胞中。 结论:这项工作清楚地证明了生物催化病毒样颗粒作为临床应用中与医学相关的酶递送载体的潜力。
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