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Design of functional cationic microgels as conjugation scaffolds

机译：缀合支架的功能阳离子微凝胶设计

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摘要

We present the development and detailed characterization of a range of amine functionalized microgels for utilization in conjugation reactions. Cationic N-isopropylmethacrylamide (NIPMAm) based microgels were synthesized through copolymerization with a primary amine containing monomer, N-(3-aminopropyl) methacrylamide hydrochloride (APMA). A range of synthesis conditions and monomer feed ratios generated microgels of diverse architectures, in different size ranges and with varying amounts of incorporated primary amines. The efficiency of amine incorporation was quantified using a fluorescence-based assay in order to determine the potential applicability of these particles for controlled bioconjugation reactions. The pH responsivity of all microgels was studied via dynamic light scattering and their height profile was investigated through atomic force microscopy following deposition on a functionalized flat substrate. Tunable resistive pulse sensing was employed to characterize microgels with respect to their number densities and molecular weights. These microgels were then conjugated to two dyes, malachite green and rose bengal with the purpose of investigating accessibility of primary amine groups for conjugation reactions. The microgel-dye constructs were then analyzed for dye content/ microgel. Finally, the viability of NIH 3T3 fibroblasts incubated in the presence of varying concentrations of non-conjugated and dye conjugated microgels was studied. Confocal imaging revealed low cellular toxicity under conditions of incubation with low concentrations of microgel-dye conjugates, which is promising for eventual utilization of these constructs in bioimaging applications.
机译：我们介绍了一系列胺官能化微凝胶的开发和详细表征，以利用共轭反应。通过与含有伯胺单体，N-（3-氨基丙基）甲基丙烯酰胺（APMA）的丙酰胺（APMA）共聚合成阳离子N-异丙基甲基丙烯酰胺（NIPMAM）的微凝胶。一系列合成条件和单体饲料比产生多种架构的微凝胶，不同的尺寸范围和不同的掺入伯胺的不同量。使用基于荧光的测定来定量胺掺入的效率，以确定这些颗粒对受控生物缀合物的潜在适用性。通过动态光散射研究所有微凝胶的pH响应度，并通过在官能化的扁平基板上沉积后通过原子力显微镜研究其高度分布。使用可调谐电阻脉冲感测来表征相对于它们数量密度和分子量的微凝胶。然后将这些微凝胶缀合至两种染料，孔雀石绿色和玫瑰瓣，目的是研究伯胺基团的可及性缀合反应。然后分析微凝胶 - 染料构建体用于染料含量/微凝胶。最后，研究了在不同浓度的非共轭和染料共轭微凝胶存在下孵育的NIH 3T3成纤维细胞的活力。共聚焦成像在与低浓度的微凝胶 - 染料缀合物温育条件下显示出低细胞毒性，这是有希望最终利用这些构建体在生物分析应用中。

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  • 来源
    《RSC Advances》 |2016年第38期|共13页
  • 作者

    Kodlekere Purva; Cartelle Anabel Liyen; Lyon L. Andrew;

  • 作者单位

    Georgia Inst Technol Sch Chem &

    Biochem Atlanta GA 30332 USA;

    Georgia Inst Technol Sch Chem &

    Biochem Atlanta GA 30332 USA;

    Georgia Inst Technol Sch Chem &

    Biochem Atlanta GA 30332 USA;

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  • 正文语种 eng
  • 中图分类 化学;
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