Controlling Ligand Surface Density on Streptavidin-Magnetic Particles by a Simple, Rapid, and Reliable Chemiluminescent Test,Bioconjugate Chemistry

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Controlling Ligand Surface Density on Streptavidin-Magnetic Particles by a Simple, Rapid, and Reliable Chemiluminescent Test
Bioconjugate Chemistry ( IF 4.0 ) Pub Date : 2018-07-10 00:00:00 , DOI: 10.1021/acs.bioconjchem.8b00347
Diego Bouzas-Ramos _{1,

2} , Laura Trapiella-Alfonso _{1,

3} , Kelly Pons _{1,

3} , Jorge Ruiz Encinar ₂ , José M. Costa-Fernández ₂ , Vassilis Tsatsaris ₃ , Nathalie Gagey-Eilstein _{1,

3}

Affiliation

The use of functionalized magnetic particles is increasing because they simplify the analytical process and yield promising results in a wide range of applications. Particularly, streptavidin-coated magnetic beads offer the possibility of rapid and very efficient grafting of biomolecules. Unfortunately, current methods to monitor and compute this grafting process are cumbersome and scarce. We describe herein a simple, rapid, and reliable chemiluminescent assay we have developed to check the grafting rate of functionalized magnetic beads. The power of the assay also relies on its ability to predict the amount of ligands required to obtain a precise grafting rate. In addition, results were correlated with a more general parameter in material functionalization characterization like surface ligand density. Finally, the assay was validated for a wide variety of biotinylated biomolecule sizes, ranging from small molecules (around 200 Da) to antibodies (around 150 kDa). This approach will allow a precise quantification and prediction of the functionalization of magnetic particles that is of enormous importance for quality control in many applications.

中文翻译：

通过简单，快速和可靠的化学发光测试控制链霉亲和素磁性颗粒上的配体表面密度

功能化磁性颗粒的使用正在增加，因为它们简化了分析过程并在广泛的应用中产生了令人鼓舞的结果。特别地，链霉抗生物素蛋白包被的磁珠提供了快速和非常有效的生物分子移植的可能性。不幸的是，用于监视和计算该嫁接过程的当前方法麻烦且稀缺。我们在这里描述了一种简单，快速和可靠的化学发光测定方法，我们已经开发出该方法来检查功能化磁珠的接枝率。测定的能力还取决于其预测获得精确接枝率所需配体数量的能力。此外，结果与材料功能化表征中更通用的参数（如表面配体密度）相关。最后，该方法针对各种生物素化生物分子大小进行了验证，从小分子（约200 Da）到抗体（约150 kDa）不等。这种方法将允许对磁性粒子的功能进行精确的定量和预测，这对于许多应用中的质量控制至关重要。

更新日期：2018-07-10

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