Biotechnology Group, Faculty of Chemical Engineering, Tarbiat Modares University, Tehran, Iran.
Int J Pharm. 2013 Apr 15;447(1-2):62-9. doi: 10.1016/j.ijpharm.2013.02.043. Epub 2013 Feb 27.
Human serum albumin (HSA) nanoparticles represent an attractive strategy for active targeting of therapeutics into tumor cells due to the presence of superficial functional groups. HER2 is highly expressed in a significant proportion of cancers and monoclonal antibodies (mAbs) directed against HER2 hold great promise for effective therapy. Herein, covalent coupling of a novel mAb (1F2) directed against the extracellular domain of HER2 to the surface of HSA nanoparticles was evaluated to obtain nanoparticles with highest cellular uptake. HER2 reactivity of 1F2-conjugated nanoparticles produced under different conditions was screened by an indirect ELISA and flow cytometry techniques. Monoclonal antibody thiolation with 100-fold molar excess of 2-iminothiolane and the ratio of 10:1 for the thiolated 1F2 (μg) to PEGylated nanoparticles (mg), were optimum for the attachment process. Under this condition, 23±4% of 1F2 was conjugated to nanoparticles. The flow cytometry results show that 1F2-modified nanoparticles interact with nearly all HER2 receptors on the surface of BT474 cells. In addition, no cellular uptake was observed on MCF7 cells. In vitro analyses showed no significant cytotoxicity of produced system against BT474 cells. Therefore, 1F2-attached HSA nanoparticles represent a potential delivery system for targeted transport of therapeutic agents into HER2-positive tumor cells.
人血清白蛋白（HSA）纳米颗粒由于其表面存在的功能基团，代表了一种将治疗药物主动靶向肿瘤细胞的有吸引力的策略。HER2 在很大一部分癌症中高度表达，针对 HER2 的单克隆抗体（mAb）为有效治疗提供了巨大的希望。在此，通过间接 ELISA 和流式细胞术技术筛选了针对 HER2 细胞外结构域的新型 mAb（1F2）与 HSA 纳米颗粒表面的共价偶联，以获得具有最高细胞摄取率的纳米颗粒。不同条件下制备的 1F2 偶联纳米颗粒的 HER2 反应性通过间接 ELISA 和流式细胞术技术进行筛选。用 2-亚氨基硫代乙烷的 100 倍摩尔过量进行单克隆抗体巯基化，以及用 PEG 化纳米颗粒（mg）的 10:1 的比例（μg）进行巯基化 1F2（μg），是附着过程的最佳条件。在此条件下，23±4%的 1F2 被偶联到纳米颗粒上。流式细胞术结果表明，1F2 修饰的纳米颗粒与 BT474 细胞表面几乎所有的 HER2 受体相互作用。此外，在 MCF7 细胞上未观察到细胞摄取。体外分析表明，所产生的系统对 BT474 细胞没有明显的细胞毒性。因此，1F2 结合的 HSA 纳米颗粒代表了将治疗剂靶向递送至 HER2 阳性肿瘤细胞的潜在递送系统。
