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聚羟基脂肪酸(PHA)颗粒表面结合蛋白Pha P具有与疏水性高分子材料表面紧密结合的能力,本研究将EGFR靶向多肽(ETP)与PhaP进行融合表达,构建了ETP-PhaP融合蛋白表达的重组工程菌Escherichia coli BL21(DE3)(pPI-ETP-P)。经对工程菌株的诱导表达及ETP-PhaP融合蛋白的纯化后,通过PhaP蛋白介导能够有效地将ETP-PhaP融合蛋白修饰于3-羟基丁酸-3-羟基己酸共聚酯(PHBHHx)纳米微球表面,构建成为具有EGFR靶向作用的药物递送载体。分别检测宫颈癌细胞系SiHa(EGFR高表达)和CaSKi(EGFR低表达)对ETP-PhaP修饰的PHBHHx纳米药物载体和未经修饰的纳米药物载体的吞噬情况。结果显示,纯化的ETP-PhaP融合蛋白能够很好地吸附于PHBHHx颗粒的表面,经ETP-PhaP融合蛋白修饰的PHBHHx纳米药物载体对EGFR高表达的宫颈癌Si Ha细胞的靶向效果强于EGFR低表达的CaSKi细胞系。这一结果表明了PhaP介导的PHBHHx纳米微球表面EGFR靶向多肽修饰具有简便、高效的优势,为疏水性纳米药物载体表面功能多肽修饰提供了一种新策略。
The PHA particle surface binding protein Pha P has the ability to bind tightly to the hydrophobic polymer surface. In this study, EGFP targeting peptide (ETP) was fused with PhaP to construct the recombinant ETP-PhaP fusion protein Recombinant engineering bacteria Escherichia coli BL21 (DE3) (pPI-ETP-P). After induced by engineering strain and purification of ETP-PhaP fusion protein, ETP-PhaP fusion protein can be effectively modified into 3-hydroxybutyrate-3-hydroxycaproic acid copolyester (PHBHHx) The surface of nanospheres was constructed as a drug delivery vehicle with EGFR targeting effect. Phagocytosis of cervical cancer cell lines SiHa (EGFR overexpression) and CaSKi (EGFR overexpression) were examined for ETP-PhaP-modified PHBHHx nanomedicine carriers and unmodified nanophase carriers, respectively. The results showed that the purified ETP-PhaP fusion protein could adsorb well on the surface of PHBHHx particles. The targeting effect of PHBHHx nano-drug carrier modified by ETP-PhaP fusion protein on Si Ha cells with high EGFR expression was stronger than that of EGFR Low expression of CaSKi cell line. This result shows that the Phap-mediated modification of EGFR-targeting peptides on the surface of PHBHHx nanospheres has the advantages of simplicity and efficiency, and provides a new strategy for the modification of functional polypeptides on the surface of hydrophobic nanocarriers.