Synthesis of a Functional Metal-Chelating Polymer and Steps toward Quantitative Mass Cytometry Bioassays

被引：106

作者：

Majonis, Daniel ^{[1
]}

Herrera, Isaac ^{[1
]}

Ornatsky, Olga ^{[1
]}

Schulze, Maren ^{[1
]}

Lou, Xudong ^{[1
]}

Soleimani, Mohsen ^{[1
]}

Nitz, Mark ^{[1
]}

Winnik, Mitchell A. ^{[1
]}

机构：

[1] Univ Toronto, Dept Chem, Toronto, ON M5S 3H6, Canada

来源：

ANALYTICAL CHEMISTRY | 2010年 / 82卷 / 21期

基金：

加拿大自然科学与工程研究理事会;

关键词：

DIETHYLENETRIAMINEPENTAACETIC ACID; POLY(ACRYLIC ACID); NANOPARTICLES; SPECTROMETRY; LYMPHOCYTES; BIOMARKERS; ANTIBODY; BEHAVIOR; BINDING;

D O I：

10.1021/ac101901x

中图分类号：

O65 [分析化学];

学科分类号：

070302 ; 081704 ;

摘要：

We describe the synthesis and characterization of metal-chelating polymers with a degree of polymerization of 67 and 79, high diethylenetriaminepentaacetic acid (DTPA) functionality, M-w/M-n <= 1.17, and a maleimide as an orthogonal functional group for conjugation to antibodies. The polymeric disulfide form of the DPn = 79 DTPA polymer was analyzed by thermogravimetric analysis to determine moisture and sodium-ion content and by isothermal titration calorimetry (ITC) to determine the Gd3+ binding capacity. These results showed each chain binds 68 +/- 7 Gd3+ per chain. Secondary goat antimouse IgG was covalently labeled with the maleimide form of the DTPA polymer (DPn = 79) carrying Tb-159. Conventional ICPMS analysis of this conjugate showed each antibody carried an average of 161 +/- 4 Tb-159 atoms. This result was combined with the ITC result to show there are an average of 2.4 +/- 0.3 polymer chains attached to each antibody. Eleven monoclonal primary antibodies were labeled with different lanthanide isotopes using the same labeling methodology. Single cell analysis of whole umbilical cord blood stained with a mixture of 11. metal-tagged antibodies was performed by mass cytometry.

引用

页码：8961 / 8969

页数：9

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