Solid-phase synthesis of molecularly imprinted polymer nanolabels: Affinity tools for cellular bioimaging of glycans
Solid-phase synthesis of molecularly imprinted polymer nanolabels: Affinity tools for cellular bioimaging of glycans
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DOI：
10.1038/s41598-019-40348-5
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发表时间：
2019-03-08
影响因子：
4.6
通讯作者：
Haupt, Karsten
中科院分区：
文献类型：
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作者：
Rangel, Paulina X. Medina;Lacief, Sylvain;Haupt, Karsten
透明质酸（HA）是一种糖胺聚糖，在健康和疾病中起着许多作用，并且是某些癌症的关键生物标志物。因此，在早期阶段，通过组织化学方法检测其是重要的。然而，细胞内HA可以被其他HA结合大分子掩蔽，使得其可视化在某种程度上有问题。我们发现，荧光分子印迹聚合物纳米凝胶（MIP-NPs），可以定位和检测细胞内的HA。通过在玻璃珠（GB）上的固相合成来合成MIP-NP。通过点击化学将模板分子葡萄糖醛酸的叠氮衍生物固定在末端炔基上，使GB官能化。通过异头碳的固定化使模板的羧基部分游离，以实现与基于苯甲脒的功能单体的强化学计量静电相互作用，从而赋予MIP-NP选择性识别。由于模板的两点取向，所得的MIP-NP被赋予改进的结合位点均一性和特异性，使人联想到单克隆抗体。然后将这些合成抗体用于探测和染色人表皮细胞上的HA，其中葡萄糖醛酸是HA的亚结构（表位）。它们优异的灵敏度、小尺寸和水相容性使得MIP-NP能够可视化HA，如通过共聚焦荧光显微照片所证明的。
Hyaluronic acid (HA) is a glycosaminoglycan that plays many roles in health and disease and is a key biomarker of certain cancers. Therefore, its detection at an early stage, by histochemical methods, is of importance. However, intracellular HA can be masked by other HA-binding macromolecules, rendering its visualization somehow problematic. We show that fluorescent molecularly imprinted polymer nanogels (MIP-NPs), can localize and detect intracellular HA. MIP-NPs were synthesized by solid-phase synthesis on glass beads (GBs). GBs were functionalized with terminal alkyne groups on which an azide derivative of the template molecule glucuronic acid was immobilized via click chemistry. Immobilization via the anomeric carbon left the template's carboxyl moiety free to enable strong stoichiometric electrostatic interactions with a benzamidine-based functional monomer, to confer selective recognition to the MIP-NPs. Due to the two-point orientation of the template, the resulting MIP-NPs were endowed with improved binding site homogeneity and specificity, reminiscent of monoclonal antibodies. These synthetic antibodies were then applied for probing and staining HA, of which glucuronic acid is a substructure (epitope), on human epidermal cells. Their excellent sensitivity, small size and water compatibility, enabled the MIP-NPs to visualize HA, as evidenced by confocal fluorescence micrographs.