ELISA-mimic screen for synthetic polymer nanoparticles with high affinity to target proteins.
ELISA-mimic screen for synthetic polymer nanoparticles with high affinity to target proteins.
复制标题
DOI：
10.1021/bm300986j
复制
发表时间：
2012-09-10
影响因子：
6.2
通讯作者：
Shea, Kenneth J.
中科院分区：
文献类型：
--
作者：
Yonamine, Yusuke;Hoshino, Yu;Shea, Kenneth J.
对蛋白质靶点表现出高亲和力的合成聚合物纳米粒子（NP）作为蛋白质捕获剂或抗体（“塑料抗体”）的功能替代品在医学和生物技术应用中具有巨大的潜力。在本研究中，我们将免疫学测定（酶联免疫吸附测定：ELISA）修改为高通量筛选方法，以选择与目标蛋白具有高亲和力的纳米颗粒。选择组蛋白和纤维蛋白原作为靶蛋白来证明这一概念。选择过程利用了由功能单体组合构建的生物素化纳米粒子库。筛选鉴定出具有独特官能团组成的纳米粒子，这些纳米粒子对组蛋白或纤维蛋白原表现出高亲和力。蛋白质亲和力随着 NP 中官能团的性质和数量的变化而变化，为蛋白质-NP 结合的主要决定因素提供了化学见解。通过改变 NP 浓度，使用 ELISA 模拟测定法对 NP 亲和力进行半定量。发现筛选结果与基于溶液的测定结果相关。这种利用生物素化纳米粒子的筛选系统是优化功能单体组合物的通用方法，可用于快速搜索对目标生物大分子具有高（或低）亲和力的合成聚合物。
Synthetic polymer nanoparticles (NPs) that display high affinity to protein targets have significant potential for medical and biotechnological applications as protein capture agents or functional replacements of antibodies (“plastic antibodies”). In this study, we modified an immunological assay (enzyme-linked immunosorbent assay: ELISA) into a high-throughput screening method to select nanoparticles with high affinity to target proteins. Histone and fibrinogen were chosen as target proteins to demonstrate this concept. The selection process utilized a biotinylated NP library constructed with combinations of functional monomers. The screen identified NPs with distinctive functional group compositions that exhibited high affinity to either histone or fibrinogen. The variation of protein affinity with changes in the nature and amount of functional groups in the NP provided chemical insight into the principle determinants of protein-NP binding. The NP affinity was semi-quantified using the ELISA-mimic assay by varying the NP concentrations. The screening results were found to correlate with solution-based assay results. This screening system utilizing a biotinalated NP is general approach to optimize functional monomer compositions and can be used to rapidly search for synthetic polymers with high (or low) affinity for target biological macromolecules.
登录
查看更多内容
影响因子：
--
作者：
Schrader, Thomas;Koch, Sebastian
通讯作者：
Koch, Sebastian
DOI：
10.1016/0005-2795(80)90287-1
发表时间：
1980-01-01
期刊：
BIOCHIMICA ET BIOPHYSICA ACTA
影响因子：
--
作者：
RAMONDABAN, J;DOLCAGUASCH, M
通讯作者：
DOLCAGUASCH, M
影响因子：
15
作者：
Renner, C;Piehler, J;Schrader, T
通讯作者：
Schrader, T
影响因子：
3.4
作者：
Zhou, XC;Zhou, JZ
通讯作者：
Zhou, JZ
影响因子：
4.7
作者：
Han, G;Chari, NS;Rotello, VM
通讯作者：
Rotello, VM