Fan Fan, Ma Jiangang, Lu Xiaoyun, Niu Teng, Wu Zhimin
Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, Shaanxi, China.
Sheng Wu Gong Cheng Xue Bao. 2017 Jun 25;33(6):1028-1036. doi: 10.13345/j.cjb.160454.
PHA granule binding protein phasin (PhaP) has a high affinity for hydrophobic materials and can bind to hydrophobic polymers via strong hydrophobic interaction. In this study, an EGFR-targeting peptide (ETP) was fused with PhaP and the fusion protein ETP-PhaP was produced in recombinant Escherichia coli BL21 (DE3) (pPI-ETP-P) and then purified by Ni affinity purification. The tumor targeting PHBHHx nanoparticles were developed based on PhaP mediated ETP immobilization and the cellular uptake of the ETP-PhaP modified PHBHHx NPs and none modified PHBHHx NPs by cervical cancer cell lines SiHa (EGFR over expressed) and CaSKi (EGFR low expressed) were analyzed. The purified ETP-PhaP could be adsorbed onto the hydrophobic surface of PHBHHx NPs. The ETP-PhaP modified PHBHHx NPs could target to EGFR over expressed cervical cancer cells SiHa more efficiently than to the EGFR low expressed CaSKi cells. These results demonstrated the advantage in effectiveness and convenience of PhaP mediated ETP adsorption on PHBHHx nanoparticles, providing a novel strategy for hydrophobic nanocarrier surface modification.
聚羟基脂肪酸酯（PHA）颗粒结合蛋白相蛋白（PhaP）对疏水材料具有高亲和力，并且可以通过强疏水相互作用与疏水聚合物结合。在本研究中，将表皮生长因子受体（EGFR）靶向肽（ETP）与PhaP融合，并在重组大肠杆菌BL21（DE3）（pPI - ETP - P）中产生融合蛋白ETP - PhaP，然后通过镍亲和纯化进行纯化。基于PhaP介导的ETP固定化开发了肿瘤靶向性聚（3 - 羟基丁酸酯 - 共 - 3 - 羟基己酸酯）（PHBHHx）纳米颗粒，并分析了ETP - PhaP修饰的PHBHHx纳米颗粒和未修饰的PHBHHx纳米颗粒被宫颈癌细胞系SiHa（EGFR高表达）和CaSki（EGFR低表达）的细胞摄取情况。纯化的ETP - PhaP可以吸附到PHBHHx纳米颗粒的疏水表面。ETP - PhaP修饰的PHBHHx纳米颗粒对EGFR高表达的宫颈癌细胞SiHa的靶向效率高于EGFR低表达的CaSki细胞。这些结果证明了PhaP介导的ETP吸附在PHBHHx纳米颗粒上在有效性和便利性方面的优势，为疏水纳米载体表面修饰提供了一种新策略。
