Chemoenzymatic strategy for the synthesis of site-specifically labeled immunoconjugates for multimodal PET and optical imaging.
Chemoenzymatic strategy for the synthesis of site-specifically labeled immunoconjugates for multimodal PET and optical imaging.
复制标题
DOI：
10.1021/bc500499h
复制
发表时间：
2014-12-17
影响因子：
4.7
通讯作者：
Lewis, Jason S.
中科院分区：
文献类型：
--
作者：
Zeglis, Brian M.;Davis, Charles B.;Abdel-Atti, Dalya;Carlin, Sean D.;Chen, Aimei;Aggeler, Robert;Agnew, Brian J.;Lewis, Jason S.
正电子发射断层扫描 (PET) 和光学成像 (OI) 的互补性质激发了人们对开发可用于癌症诊断、分期和手术治疗的多模式 PET/OI 探针的兴趣。由于其高选择性和亲和力，抗体已成为开发混合 PET/OI 试剂的有前途的平台。然而，许多生物缀合反应缺乏特异性可能会威胁免疫反应性并导致结构不明确。为了解决这个问题，我们开发了一种化学酶策略来构建多模式 PET/OI 免疫缀合物，这些免疫缀合物已在重链聚糖上进行了位点特异性标记。该方法由四个步骤组成：（1）酶法去除重链聚糖上的末端半乳糖残基； (2) 将带有叠氮化物的半乳糖 (GalNAz) 残基酶促掺入重链聚糖中； (3)菌株促进的螯合剂和荧光团修饰的二苯并环辛炔与叠氮修饰的糖的点击缀合； (4)免疫缀合物的放射性标记。为了进行概念验证，使用结直肠癌靶向抗体 huA33、螯合剂去铁胺 (DFO)、正电子发射放射性金属 89Zr 和近红外荧光染料 Alexa Fluor 680 创建了一个模型系统。生物缀合策略稳健且可重复，能够可靠地产生用 89Zr、Alexa Fluor 标记的明确且具有免疫反应性的缀合物。 680，或两个报告器的轻松且精确调整的混合。在体内 PET 和荧光成像实验中，混合 89Zr 和 Alexa Fluor 680 标记的 huA33 缀合物在携带表达 A33 抗原的 SW1222 结直肠癌异种移植物的无胸腺裸鼠中表现出高水平的特异性摄取 (> 45% ID/g)。
The complementary nature of positron emission tomography (PET) and optical imaging (OI) has fueled increasing interest in the development of multimodal PET/OI probes that can be employed during the diagnosis, staging, and surgical treatment of cancer. Due to their high selectivity and affinity, antibodies have emerged as promising platforms for the development of hybrid PET/OI agents. However, the lack of specificity of many bioconjugation reactions can threaten immunoreactivity and lead to poorly defined constructs. To circumvent this issue, we have developed a chemoenzymatic strategy for the construction of multimodal PET/OI immunoconjugates that have been site-specifically labeled on the heavy chain glycans. The methodology consists of four steps: (1) the enzymatic removal of the terminal galactose residues on the heavy chain glycans; (2) the enzymatic incorporation of azide-bearing galactose (GalNAz) residues into the heavy chain glycans; (3) the strain-promoted click conjugation of chelator- and fluorophore-modified dibenzocyclooctynes to the azide-modified sugars; and (4) the radiolabeling of the immunoconjugate. For proof-of-concept, a model system was created using the colorectal cancer-targeting antibody huA33, the chelator desferrioxamine (DFO), the positron-emitting radiometal 89Zr, and the near-infrared fluorescent dye Alexa Fluor 680. The bioconjugation strategy is robust and reproducible, reliably producing well-defined and immunoreactive conjugates labeled with 89Zr, Alexa Fluor 680, or an easily and precisely tuned mixture of the two reporters. In in vivo PET and fluorescence imaging experiments, a hybrid 89Zr- and Alexa Fluor 680-labeled huA33 conjugate displayed high levels of specific uptake (>45% ID/g) in athymic nude mice bearing A33 antigen-expressing SW1222 colorectal cancer xenografts.
登录
查看更多内容
影响因子：
62.1
作者：
Louie, Angelique Y.
通讯作者：
Louie, Angelique Y.
影响因子：
14.8
作者：
Cohen, Ruth;Vugts, Danielle J.;van Dongen, Guus A. M. S.
通讯作者：
van Dongen, Guus A. M. S.
影响因子：
--
作者：
Seibold U;Wängler B;Schirrmacher R;Wängler C
通讯作者：
Wängler C
影响因子：
16.6
作者：
Ning, Xinghai;Guo, Jun;Boons, Geert-Jan
通讯作者：
Boons, Geert-Jan
影响因子：
4.7
作者：
Hussain, Ahmad Fawzi;Kampmeier, Florian;Barth, Stefan
通讯作者：
Barth, Stefan