Characterization methods for studying protein adsorption on nano-polystyrene beads
Characterization methods for studying protein adsorption on nano-polystyrene beads
复制标题
DOI：
10.1016/j.chroma.2019.460383
复制
发表时间：
2019-11-22
影响因子：
4.1
通讯作者：
Calzolai, Luigi
中科院分区：
文献类型：
--
作者：
Contado, Catia;Mehn, Dora;Calzolai, Luigi
这项工作是关于使用聚苯乙烯(PS)纳米颗粒作为生物分析特定相互作用的底物。使用不同的技术对100 nm和196 nm的PS纳米颗粒进行精确表征，然后在它们上覆盖一层抗E型免疫球蛋白(AIGE)的抗体，赋予颗粒特定的功能。通过离心颗粒分离(CPS)和离心场流动分馏(CF3)实验对AIGE吸附层的形成进行了监测，以确定其尺寸变化和吸附质量。颗粒大小也用DLS测量，DLS既可作为独立仪器使用，也可与CF3(CF3-DLS)耦合使用。从CPS和CF3-DLS测量中获得的补充信息使我们能够估计Aige壳的密度。表面固定的蛋白质完全保持了它们的活性，功能化的PS-AIGE颗粒与特意制备的悬浮液中分散的E型免疫球蛋白(IgE)之间的反应证明了这一点。(C)2019年提交人。爱思唯尔出版公司(Elsevier B.V.)
This work is dealing with the use of polystyrene (PS) nanoparticles as substrates for bioanalytical specific interactions. Different techniques were used for the accurate characterization of the PS nanoparticles of 100 nm and 196 nm before coating them with a layer of antibodies against immunoglobulins of type E (aIgE), giving to the particle a specific functionality. The formation of the aIgE adsorbed layer was monitored using centrifugal particle separation (CPS) and centrifugal field flow fractionation (CF3) experiments, which allowed to determine the size changes and the adsorbed mass. Particle sizes were also measured with DLS, used both as stand-alone instrument and coupled to CF3 (CF3-DLS). The complementary information obtained from the CPS and CF3-DLS measurements allowed the estimation of the density of the aIgE shell. The proteins immobilized at the surface fully retained their activity, as proven by the reactions between the functionalized PS-aIgE particles and immunoglobulins of type E (IgE) dispersed in suspensions prepared on purpose. (C) 2019 The Authors. Published by Elsevier B.V.