当前位置: X-MOL 学术Soft Matter › 论文详情
Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)
Surface plasmon resonance study for a reliable determination of the affinity constant of multivalent grafted beads
Soft Matter ( IF 2.9 ) Pub Date : 2021-07-12 , DOI: 10.1039/d1sm00591j
Giuseppina Simone 1
Affiliation  

In this study, galactose-grafted beads were prepared using the main design principle of the cluster effect. Galactose was chosen as the sugar for investigation because it acts as the main building block of long glycan chains and because a simple and fast protocol is still required for its immobilization. For the analysis, the lectin, ligand of the galactose, was immobilized on a gold plasmonic substrate. After preliminary characterization of the galactose-grafted beads, the investigation of the surface plasmon surface behavior of the system was carried out, for studying the affinity constant of the multivalent beads. The results of steady-state and of the kinetics analysis evidenced a higher affinity of the galactose-grafted beads over the beadless galactose solution. For the association kinetics analysis, a Langmuir isotherm was applied to the data. The analysis of the rate of dissociation evidenced the most important differences between the two samples, based on the more difficult release of the galactose-grafted beads during washing. To confirm the influence of the glycoside cluster effect, a low-density lectin substrate was tested, and the results evidenced that the characteristic size of the molecules determines a threshold for the cluster density. The calculated detection limit and dissociation constants were 3.5 μM and 40.2 μM, respectively. Considering those results, the evaluation of the affinities toward the receptors depends on the cluster density and then, it should be designed for mimicking the biological samples.

中文翻译:

用于可靠测定多价接枝珠的亲和常数的表面等离子体共振研究

在本研究中,使用簇效应的主要设计原理制备了半乳糖接枝珠。选择半乳糖作为研究的糖是因为它是长聚糖链的主要组成部分,并且因为它的固定仍然需要一个简单快速的方案。为了进行分析,半乳糖的配体凝集素被固定在金等离子体基质上。在对半乳糖接枝珠进行初步表征后,对系统的表面等离子体表面行为进行了研究,以研究多价珠的亲和常数。稳态和动力学分析的结果证明半乳糖接枝珠比无珠半乳糖溶液具有更高的亲和力。对于缔合动力学分析,Langmuir 等温线应用于数据。基于半乳糖接枝珠在洗涤过程中更难释放,解离速率分析证明了两个样品之间最重要的差异。为了确认糖苷簇效应的影响,测试了低密度凝集素底物,结果证明分子的特征大小决定了簇密度的阈值。计算出的检测限和解离常数分别为 3.5 μM 和 40.2 μM。考虑到这些结果,对受体亲和力的评估取决于簇密度,然后,它应该被设计为模拟生物样品。基于在洗涤过程中更难释放半乳糖接枝的珠子。为了确认糖苷簇效应的影响,测试了低密度凝集素底物,结果证明分子的特征大小决定了簇密度的阈值。计算出的检测限和解离常数分别为 3.5 μM 和 40.2 μM。考虑到这些结果,对受体亲和力的评估取决于簇密度,然后,它应该被设计为模拟生物样品。基于在洗涤过程中更难释放半乳糖接枝的珠子。为了确认糖苷簇效应的影响,测试了低密度凝集素底物,结果证明分子的特征大小决定了簇密度的阈值。计算出的检测限和解离常数分别为 3.5 μM 和 40.2 μM。考虑到这些结果,对受体亲和力的评估取决于簇密度,然后,它应该被设计为模拟生物样品。结果证明分子的特征大小决定了簇密度的阈值。计算出的检测限和解离常数分别为 3.5 μM 和 40.2 μM。考虑到这些结果,对受体亲和力的评估取决于簇密度,然后,它应该被设计为模拟生物样品。结果证明分子的特征大小决定了簇密度的阈值。计算出的检测限和解离常数分别为 3.5 μM 和 40.2 μM。考虑到这些结果,对受体亲和力的评估取决于簇密度,然后,它应该被设计为模拟生物样品。
更新日期:2021-07-12
down
wechat
bug