三维纳米结构稳定磷脂聚合物表面用于高灵敏度免疫分析。

Stabilization of phospholipid polymer surface with three-dimensional nanometer-scaled structure for highly sensitive immunoassay.

机构信息

Department of Materials Engineering, School of Engineering, The University of Tokyo, 7-3-1, Hongo, Bunkyo-ku, Tokyo 113-8656, Japan.

出版信息

Colloids Surf B Biointerfaces. 2010 Jun 1;77(2):263-9. doi: 10.1016/j.colsurfb.2010.02.008. Epub 2010 Feb 11.

DOI:10.1016/j.colsurfb.2010.02.008

PMID:20197230

Abstract

A phospholipid polymer platform and an antibody as a bioaffinity ligand were used to construct a biointerface for a highly sensitive immunoassay. The platform had a nanometer-scaled particle deposition surface and it was constructed with poly[2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate (BMA)-co-p-nitrophenyloxycarbonyl poly(ethylene glycol) methacrylate (MEONP)] (PMBN) by an electrospray deposition (ESD) method. The PMBN surface could immobilize specific antibodies through covalent chemical bonding by the reaction between MEONP units and amino groups in the antibody. In addition, the PMBN could prevent nonspecific protein adsorption from an analyte. However, the nanometer-scaled structure of the PMBN lost its shape after immersion in an aqueous medium. To stabilize the nanometer-scaled structure in an aqueous medium, the PMBN was cross-linked with 1,4-butylenediamine and then heat-treated. These treatments effectively improved the stability of the nanometer-scaled structure, that is, the structure had a high porosity even after immersing in an aqueous medium. The stabilization affected the specific signal in the enzyme-linked immunosorbent assay (ELISA), that is, the specific signal in ELISA was enhanced.

摘要

一种磷脂聚合物平台和一种抗体作为生物亲和配体被用于构建高灵敏度免疫分析的生物界面。该平台具有纳米级颗粒沉积表面，通过电喷雾沉积（ESD）方法用聚[2-（甲基丙烯酰氧基）乙基磷酸胆碱（MPC）-共-正丁基甲基丙烯酸酯（BMA）-共-p-硝基苯氧羰基聚（乙二醇）甲基丙烯酸酯（MEONP）]（PMBN）构建。PMBN 表面可以通过 MEONP 单元与抗体中氨基之间的反应通过共价化学键固定特异性抗体。此外，PMBN 可以防止分析物中的非特异性蛋白质吸附。然而，PMBN 的纳米级结构在浸入水介质后会失去形状。为了在水介质中稳定纳米级结构，PMBN 用 1,4-丁二胺交联，然后进行热处理。这些处理有效地提高了纳米级结构的稳定性，即使在浸入水介质后，结构仍具有高孔隙率。这种稳定作用影响酶联免疫吸附测定（ELISA）中的特异性信号，即 ELISA 中的特异性信号得到增强。