Enzyme-Mediated Methodology for the Site-Specific Radiolabeling of Antibodies Based on Catalyst-Free Click Chemistry

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• 作者：Brian M. Zeglis ; Charles B. Davis ; Robert Aggeler ; Hee Chol Kang ; Aimei Chen ; Brian J. Agnew ; Jason S. Lewis
• 刊名：Bioconjugate Chemistry
• 出版年：2013
• 出版时间：June 19, 2013
• 年：2013
• 卷：24
• 期：6
• 页码：1057-1067
• 全文大小：327K
• 年卷期：v.24,no.6(June 19, 2013)
• ISSN：1520-4812

文摘

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 highly selective tumor uptake (67.5 卤 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.