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Enabling Indium Channels for Mass Cytometry by Using Reinforced Cyclam-Based Chelating Polylysine.
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2020-06-22 , DOI: 10.1021/acs.bioconjchem.0c00267 Laura Grenier 1 , Maryline Beyler 1 , Taunia Closson 2 , Nick Zabinyakov 2 , Alexandre Bouzekri 2 , Yefeng Zhang 3 , Jothir Mayanantham Pichaandi 3 , Mitchell A Winnik 3 , Peng Liu 2 , Olga I Ornatsky 2 , Vladimir Baranov 2 , Raphaël Tripier 1
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2020-06-22 , DOI: 10.1021/acs.bioconjchem.0c00267 Laura Grenier 1 , Maryline Beyler 1 , Taunia Closson 2 , Nick Zabinyakov 2 , Alexandre Bouzekri 2 , Yefeng Zhang 3 , Jothir Mayanantham Pichaandi 3 , Mitchell A Winnik 3 , Peng Liu 2 , Olga I Ornatsky 2 , Vladimir Baranov 2 , Raphaël Tripier 1
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The synthesis of a polylysine polymer functionalized with the previously reported astonishingly inert [In(cb-te2pa)]+ chelate was performed. A biotin end group allowed the conjugation to biotinylated beads by the intermediary of a fluorescein isothiocyanate/neutravidin receptor. High quality imaging mass cytometry trials, based on 115In detection were performed to highlight the behavior of the material. Anti-CD20 antibody was labeled by the so-obtained In(III)-modified polylysine using the biotin/neutravidin interaction. Ramos (CD20[+]) and HL-60 (CD20[−]) cell lines were costained with the In(III)-modified bioconjugate by finding the best staining conditions. Both immunofluorescence microscopy (IF-M) and mass cytometry analyses confirmed the specific binding of anti-CD20 onto Ramos cells. CyTOF histograms constructed on the 115In detection allowed us to define and to separate, with a good signal-to-noise ratio, two populations (Ramos and HL-60). The inertness of In(III)-MCP-NAv over a three-month storage period was proved by performing new functionality tests involving Jurkat cells (CD20[−]) and multiparametric trials involving the 115In channel. The results ensure a promising future use of the previously announced [In(cb-te2pa)]+ complex-based polymers for mass cytometry.
中文翻译：
通过使用增强的基于Cyclam的螯合聚赖氨酸使质谱分析中的铟通道成为可能。
进行了用先前报道的惊人惰性[In（cb-te2pa）] +螯合物官能化的聚赖氨酸聚合物的合成。生物素端基通过异硫氰酸荧光素/中性亲和素受体的介导而与生物素化的珠结合。基于115的高质量成像大规模细胞术试验在检测中进行以突出材料的行为。使用生物素/中性亲和素相互作用，通过如此获得的In（III）修饰的聚赖氨酸标记抗CD20抗体。通过寻找最佳染色条件，将Ramos（CD20 [+]）和HL-60（CD20 [-]）细胞系与In（III）修饰的生物缀合物共染色。免疫荧光显微镜（IF-M）和质谱分析均证实了抗CD20与Ramos细胞的特异性结合。在115 In检测中构建的CyTOF直方图使我们能够以良好的信噪比定义和分离两个种群（Ramos和HL-60）。通过进行涉及Jurkat细胞（CD20 [-]）的新功能测试以及涉及Jurkat细胞（CD20 [-]）的多参数试验，证明了In（III）-MCP-NAv在三个月储存期内的惰性。115在频道中。结果确保了以前宣布的[In（cb-te2pa）] +基于复杂物的聚合物在细胞计数中的应用前景广阔。
更新日期：2020-06-22
中文翻译：
通过使用增强的基于Cyclam的螯合聚赖氨酸使质谱分析中的铟通道成为可能。
进行了用先前报道的惊人惰性[In（cb-te2pa）] +螯合物官能化的聚赖氨酸聚合物的合成。生物素端基通过异硫氰酸荧光素/中性亲和素受体的介导而与生物素化的珠结合。基于115的高质量成像大规模细胞术试验在检测中进行以突出材料的行为。使用生物素/中性亲和素相互作用，通过如此获得的In（III）修饰的聚赖氨酸标记抗CD20抗体。通过寻找最佳染色条件，将Ramos（CD20 [+]）和HL-60（CD20 [-]）细胞系与In（III）修饰的生物缀合物共染色。免疫荧光显微镜（IF-M）和质谱分析均证实了抗CD20与Ramos细胞的特异性结合。在115 In检测中构建的CyTOF直方图使我们能够以良好的信噪比定义和分离两个种群（Ramos和HL-60）。通过进行涉及Jurkat细胞（CD20 [-]）的新功能测试以及涉及Jurkat细胞（CD20 [-]）的多参数试验，证明了In（III）-MCP-NAv在三个月储存期内的惰性。115在频道中。结果确保了以前宣布的[In（cb-te2pa）] +基于复杂物的聚合物在细胞计数中的应用前景广阔。
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