人重链铁蛋白偶联HER2抗体的纯化及其在成像中的初步应用  

Purification of Human Heavy Chain Ferritin Coupled HER2 Antibody and its Preliminary Application in Imaging

作　　者：殷惠 陈龙 张文远 朱新杰 许诺 王婷婷[1,2] 盛复庚 YIN Hui;CHEN Long;ZHANG Wen-Yuan;ZHU Xin-Jie;XU Nuo;WANG Ting-Ting;SHENG Fu-Geng(Clinical College of 307th Hospital of PLA,Anhui Medical University,Beijing 100071,China;Department of Radiology,Southern Hospital of the Fifth Medical Center of PLA General Hospital,Beijing 100071,China;College of Chemistry and Molecular Engineering,Peking University,Beijing 100871,China)

机构地区：[1]安徽医科大学解放军307临床学院,北京100071 [2]解放军总医院第五医学中心南院区放射科,北京100071 [3]北京大学化学与分子工程学院,北京100871

出　　处：《生物技术通讯》2019年第2期235-241,共7页Letters in Biotechnology

基　　金：国家自然科学基金(21575161)

摘　　要：目的:利用临床常用抗体将人重链铁蛋白(HFn)纳米材料更好地靶向肿瘤,以期改善肿瘤的靶向成像或药物治疗,促进蛋白类纳米材料向临床转化。方法:在大肠杆菌内表达纯化得到HFn,以抗人表皮生长因子受体2抗体(anti-HER2)作为模型抗体,用双功能交联剂马来酰亚胺-聚乙二醇-琥珀酰亚胺琥珀酸酯(Mal-PEG-NHS)将HFn与anti-HER2偶联形成HFn/anti-HER2复合物;标记荧光后,观察其不同时间内在HER2阳性乳腺癌荷瘤小鼠内的分布,判断其作为靶向肿瘤药物的可能性。结果:通过尺寸排阻凝胶色谱纯化的靶向复合物HFn/anti-HER2保留典型的对称球状结构,粒径大小为16.88±0.49 nm,较HFn粒径7.11±0.1 nm增加;HFn/anti-HER2组在肿瘤部位的荧光强度是对照组HFn的3倍,说明可以通过偶联抗体提升HFn的靶向性。结论:构建了人重链铁蛋白与抗体的复合物,提升了铁蛋白的靶向性。抗体铁蛋白复合物为铁蛋白类靶向成像及治疗药物的开发提供了新的思路。Objective:To improve tumor targeting ability of human heavy chain ferritin(HFn)nanomaterials,HFn was conjugated with commercial antibodies in clinical practice,which is expected to improve the target imaging or therapy of tumors and promote the protein-based nanomaterials into clinic in the future.Methods:HFn was expressed and purified in E.coli.Anti-human epidermal growth factor receptor-2 antibody(anti-HER2)was used as model antibody to couple with HFn and form a covalent complex HFn/anti-HER2 by using Mal-PEGNHS as a heterobifunctional crosslinker.With optical imaging,the distribution of HFn/anti-HER2 in HER2-positive breast cancer-bearing mice was observed at different time points.Results:HFn/anti-HER2 was purified by size exclusion gel chromatography and retained a typical symmetrical spherical structure with a particle size of 16.88+0.49 nm,which was increased by 7.11+0.1 nm compared with the HFn particle size.The fluorescence intensity of HFn/anti-HER2 group was 3 times higher than that of the control group,indicating that the targeting of the ferritin could be enhanced by conjugating antibodies.Conclusion:The complex of HFn and anti-HER2 has been successfully conjugated,which greatly improves the targeting of HFn and provides a basic platform for the development of ferritin-targeted imaging and therapeutic drugs in the future.

关 键 词：人重链铁蛋白 抗HER2抗体 乳腺癌 肿瘤靶向 

分 类 号：R392.1[医药卫生—免疫学]