靶向巨噬细胞的纳米抗体PET分子探针^(68)Ga-NOTA-Nb119的制备方法  

Method of preparing macrophage-targeting nanobody^(68)Ga-NOTA-Nb119 as PET molecular probe

作　　者：张嘉欣 张明如 郭玉呈 庞于成 王馨怡 李璐瑶 郑芳 ZHANG Jiaxin;ZHANG Mingru;GUO Yucheng;PANG Yucheng;WANG Xinyi;LI Luyao;ZHENG Fang(Department of Biochemistry and Molecular Biology,School of Basic Medical Science,Key Laboratory of Environment and Genes Related to Disease of Ministry of Education,Xi’an Jiaotong University Health Science Center,Xi’an 710061;Department of Nuclear Medicine,Xijing Hospital of air Military Medical University,Xi’an 710032,China)

机构地区：[1]西安交通大学医学部基础医学院生物化学与分子生物学系,西安交通大学环境与疾病相关基因教育部重点实验室,陕西西安710061 [2]空军医科大学西京医院核医学科,陕西西安710032

出　　处：《西安交通大学学报（医学版）》2022年第5期752-756,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金面上项目(No.38170187);陕西省自然科学基础研究计划项目(No.2021JM-007);中国医学科学院基金资助(2021-JKCS-008)。

摘　　要：目的用^(68)Ga标记纳米抗体Nb119和NbBCⅡ10,制备靶向巨噬细胞的PET探针。方法将抗Vsig4纳米抗体Nb119和同型对照抗体BCⅡ10与螯合剂NOTA共孵育,纯化后分别与^(68)Ga进行标记。通过SDS-PAGE、MALDI-TOF方法对NOTA-Nb119进行鉴定,利用Radio-HPLC检测^(68)Ga标记后NOTA-Nb119和NOTA-BCⅡ10放化纯度。结果SDS-PAGE结果显示,成功标记的NOTA-Nb119的泳道条带比未标记纳米抗体条带出现明显的滞后性,证明标记后产物分子量变大,纳米抗体被NOTA成功修饰。^(68)Ga能成功标记纳米抗体Nb119和BCⅡ10,经Radio-HPLC检测,^(68)Ga标记的NOTA-Nb119和NOTA-BCⅡ10放化纯度均高于90%,且可稳定存在。结论^(68)Ga标记的两种纳米抗体均具有高放化纯度和高稳定性,说明本研究可推广适用于其他纳米抗体的修饰和标记,为在PET中使用纳米抗体作为分子影像探针提供了切实可行的方法。Objective To prepare positron emission computed tomography(PET)molecular probes targeting macrophages using Nb119 and BCⅡ10 labeled with ^(68)Ga nuclide.Methods To explore the labeling conditions of nanobodies with ^(68)Ga nuclides,first,the anti-Vsig4 nanobody Nb119 and isotype control antibody BCⅡ10 were incubated with NOTA and then purified to obtain the chelating agent-modified NOTA-Nb119 and NOTA-BCⅡ10.The NOTA-Nb119 and NOTA-BCⅡ10 were further incubated with ^(68)Ga.Finally,NOTA-Nb119 was identified by SDS-PAGE and MALDI-TOF methods,and the radiochemical purity was detected by radio-HPLC.Results The results of SDS-PAGE showed that the lanes of the successfully labeled NOTA-Nb119 had apparent hysteresis than the unlabeled nanobody band,which proved that the molecular weight of the labeled product was increased and NOTA successfully modified the nanobody.Subsequently,^(68)Ga nuclides could successfully label Nb119 and BCⅡ10.According to radio-HPLC detection,the radiochemical purity of NOTA-Nb119 and NOTA-BCⅡ10 labeled with ^(68)Ga was greater than 90%and remained stable.Conclusion ^(68)Ga-labeled nanobodies have high radiochemical purity and high stability,indicating that this study can be applied to other nanobodies for modification and labeling and provides a practical and feasible method for the preparation of PET using nanobodies.

关 键 词：纳米抗体 正电子发射断层扫描 NOTA ^(68)Ga标记 Radio-HPLC 

分 类 号：R392[医药卫生—免疫学]