Karir Tarveen, Hassan P A, Kulshreshtha S K, Samuel G, Sivaprasad N, Meera V
Radiopharmaceuticals Program, Board of Radiation and Isotope Technology, Department of Atomic Energy, Navi Mumbai, India.
Anal Chem. 2006 Jun 1;78(11):3577-82. doi: 10.1021/ac052032g.
The selection of an appropriate surface as a solid phase for coupling antibodies is a critical step in the development of solid-phase immunoassays. Availability of a new method of preactivating the surface of polystyrene tubes with a layer of another polymer for enhanced immobilization of antibodies seems to be promising. In this paper, we report the activation of a polystyrene surface using a layer of polyaniline and its effect on immobilizing antibodies for use as a solid phase in a T3 immunoassay. The modified surface on the polystyrene was characterized by optical absorption, X-ray photoelectron spectroscopy (XPS), and atomic force microscopy (AFM). The modified tubes were coated with antibody and evaluated for their performance in the assay and validated for radioimmunoassay of T3. AFM images of the modified surface showed an enhancement in the surface roughness (Ra of 20.2 nm), as compared to an unmodified surface (Ra of 6 nm), allowing more adsorption of antibodies to the surface. XPS revealed the presence of N (binding energy approximately 400 eV) on the modified surface, which could help the antibody molecules to bind to these preactivated (modified) tubes. The modified tubes, when coated with antibody, not only showed an increase in the binding with the radioiodinated tracer but also improved the precision of coating the antibody. The present method of activating polystyrene surfaces is simple, does not involve severe chemical treatment, and may have wide applicability to functionalize other supports for immobilizing biomolecules.
选择合适的表面作为偶联抗体的固相是固相免疫测定发展中的关键步骤。用另一层聚合物对聚苯乙烯管表面进行预活化以增强抗体固定化的新方法似乎很有前景。在本文中,我们报道了用聚苯胺层对聚苯乙烯表面进行活化及其对固定抗体用作T3免疫测定固相的影响。通过光吸收、X射线光电子能谱(XPS)和原子力显微镜(AFM)对聚苯乙烯上的改性表面进行了表征。将改性管涂上抗体,并评估其在测定中的性能,并对T3放射免疫测定进行验证。改性表面的AFM图像显示,与未改性表面(Ra为6nm)相比,表面粗糙度有所增加(Ra为20.2nm),从而使更多抗体吸附到表面。XPS显示改性表面存在N(结合能约为400eV),这有助于抗体分子与这些预活化(改性)管结合。当涂上抗体时,改性管不仅显示出与放射性碘标记示踪剂的结合增加,而且提高了抗体包被的精度。目前活化聚苯乙烯表面的方法简单,不涉及苛刻的化学处理,可能广泛适用于功能化其他用于固定生物分子的载体。