Enzyme-mediated methodology for the site-specific radiolabeling of antibodies based on catalyst-free click chemistry.
Enzyme-mediated methodology for the site-specific radiolabeling of antibodies based on catalyst-free click chemistry.
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基于无催化剂点击化学的酶介导抗体位点特异性放射性标记方法。
DOI：
10.1021/bc400122c
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发表时间：
2013-06-19
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.;Aggeler, Robert;Kang, Hee Chol;Chen, Aimei;Agnew, Brian J.;Lewis, Jason S.
已开发并验证了酶和点击化学介导的重链聚糖上抗体位点选择性放射性标记方法。为此，采用了基于前列腺特异性膜抗原靶向抗体J591、正电子发射放射性金属89 Zr和螯合剂去铁胺的模型系统。该方法由四个步骤组成：（1）去除抗体重链区上的糖以暴露末端N-乙酰葡糖胺残基;（2）将叠氮化物修饰的N-乙酰半乳糖胺单糖掺入抗体的聚糖中;（3）去铁胺修饰的二苯并环辛炔与携带叠氮化物的糖的无催化剂点击缀合;和（4）用89 Zr放射性标记螯合剂修饰的抗体。位点选择性标记方法已被证明是容易的、可重复的和稳健的，产生89 Zr标记的放射免疫缀合物，其在携带PSMA表达的皮下LNCaP异种移植物的无胸腺裸鼠中除了高选择性肿瘤摄取（67.5 ± 5.0%ID/g）和肿瘤与背景对比之外，还显示出体外高稳定性和免疫反应性（>95%）。最终，这种策略可以发挥关键作用，在新的定义明确和高度免疫反应性的放射免疫偶联物的实验室和临床的发展。
An enzyme- and click chemistry-mediated methodology for the site-selective radiolabeling of antibodies on the heavy chain glycans has been developed and validated. To this end, a model system based on the prostate specific membrane antigen-targeting antibody J591, the positron-emitting radiometal 89Zr, and the chelator desferrioxamine has been employed. The methodology consists of four steps: (1) the removal of sugars on the heavy chain region of the antibody to expose terminal N-acetylglucosamine residues; (2) the incorporation of azide-modified N-acetylgalactosamine monosaccharides into the glycans of the antibody; (3) the catalyst-free click conjugation of desferrioxamine-modified dibenzocyclooctynes to the azide-bearing sugars; and (4) the radiolabeling of the chelator-modified antibody with 89Zr. The site-selective labeling methodology has proven facile, reproducible, and robust, producing 89Zr-labeled radioimmunoconjguates that display high stability and immunoreactivity in vitro (>95%) in addition to high selective tumor uptake (67.5 ± 5.0 %ID/g) and tumor-to-background contrast in athymic nude mice bearing PSMA-expressing subcutaneous LNCaP xenografts. Ultimately, this strategy could play a critical role in the development of novel well-defined and highly immunoreactive radioimmunoconjugates for both the laboratory and clinic.
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影响因子：
7.3
作者：
Gill, Herman S.;Tinianow, Jeff N.;Marik, Jan
通讯作者：
Marik, Jan
影响因子：
4.7
作者：
Hussain, Ahmad Fawzi;Kampmeier, Florian;Barth, Stefan
通讯作者：
Barth, Stefan
影响因子：
3.7
作者：
Holland JP;Caldas-Lopes E;Divilov V;Longo VA;Taldone T;Zatorska D;Chiosis G;Lewis JS
通讯作者：
Lewis JS
影响因子：
4.7
作者：
Li, Lin;Crow, Desiree;Turatti, Fabio;Bading, James R.;Anderson, Anne-Line;Poku, Erasmus;Yazaki, Paul J.;Carmichael, Jenny;Leong, David;Wheatcroft, Michael P.;Raubitschek, Andrew A.;Hudson, Peter J.;Colcher, David;Shively, John E.
通讯作者：
Shively, John E.
DOI：
10.1073/pnas.2335201100
发表时间：
2003-12-09
影响因子：
11.1
作者：
Hang, HC;Yu, C;Bertozzi, CR
通讯作者：
Bertozzi, CR