Convenient hyaluronidase biosensors based on the target-trigger enhancing of the permeability of a membrane using an electronic balance as a readout.
Convenient hyaluronidase biosensors based on the target-trigger enhancing of the permeability of a membrane using an electronic balance as a readout.
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DOI：
10.1039/d1an00257k
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发表时间：
2021-05
期刊：
影响因子：
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通讯作者：
N. Dong;Qing Cai;Zhixin Li;Liangkui Xu;Huping Wu;Zhirong Lin;Bin Qiu;Cheng Li;Zhen Lin
中科院分区：
文献类型：
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作者：
N. Dong;Qing Cai;Zhixin Li;Liangkui Xu;Huping Wu;Zhirong Lin;Bin Qiu;Cheng Li;Zhen Lin
透明质酸酶（HAase）的变化与玻璃体结构的特异性变化有关，有必要开发简单而灵敏的HAase检测方法。在这项工作中，透明质酸(HA)-聚乙烯亚胺（PEI）水凝胶制备的薄膜首先覆盖在混合纤维素微孔膜（MCEM）上，形成HA-PEI-MCEM，然后将其应用于过滤系统。过滤膜的渗透性对一定时间内通过的水量有很大的影响，用简单的电子天平可以很容易地收集和定量测量水。HA-PEI-MCEM的低渗透率允许少量水排出。但加入能水解水凝胶中HA的HAase后，膜的通透性增加。因此，通过HA-PEI-MCEM复合膜的水量也相应增加。优化了复合膜的组成，并对加入HAase后的反应条件进行了优化。在优化条件下，5 min内取水量与HAase浓度在1.0 ~ 36 U mL-1范围内呈线性关系，检出限为0.35 U mL-1。该方法已用于玻璃体样品中HAase的检测，结果令人满意。
The change in hyaluronidase (HAase) is related to specific changes in the structure of vitreous, and it is necessary to develop simple but sensitive methods for HAase detection. In this work, a thin film fabricated from a hyaluronic acid (HA)-polyethyleneimine (PEI) hydrogel has been covered on a mixed cellulose microporous membrane (MCEM) to form a HA-PEI-MCEM firstly and it was then applied in a filtration system. The permeability of the filter membrane greatly affects the amount of water passing through within a certain time and the water can be collected and quantitatively measured with a simple electronic balance easily. The low permeability of the HA-PEI-MCEM allows a small amount of water to be drained. But after the addition of HAase, which can hydrolyze HA in the hydrogel, the permeability of the membrane increased. Therefore, the amount of water passing through the HA-PEI-MCEM composite membrane increased accordingly. The composite of the membrane, and the reaction conditions after the addition of HAase were optimized. Under the optimized conditions, the amount of water collected within 5 min showed a linear relationship with the HAase concentration in a range of 1.0-36 U mL-1 with a limit of detection of 0.35 U mL-1.The proposed method has been applied to detect HAase in vitreous samples with satisfactory results.