分子印迹二氧化硅与西尼罗河抗体模板显示特异性和选择性结合免疫分析。

Journal of biotechnology & biomaterials Pub Date : 2017-01-01 Epub Date: 2017-06-13 DOI:10.4172/2155-952x.1000260
Julio E Rincon, Fabio Diaz Santillan, Pedro M Palermo Infante, Douglas M Watts, Thomas Boland
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引用次数: 1

摘要

研究了分子印迹聚合物颗粒(MIPs)的分子印迹技术。颗粒通过溶胶-凝胶法合成,其中活性单体的相对数量与西尼罗河抗体模板的表面电荷的相对数量是互补的。合成的MIPs与亲和纯化的西尼罗河多克隆抗体(WNA)特异性结合,负载能力为80µg/mg,而MIPs吸收非特异性蛋白的负载能力为28µg/mg。结合等温线测得解离常数Kd=57.45 μM。MIPs选择性吸收的WNA比白蛋白或免疫球蛋白多27倍,而MIPs选择性吸收的WNA比非印迹颗粒(NIPs)多16倍。最后,荧光标记的mip在高结合的96孔板中孵育,预先装载模板,白蛋白或免疫球蛋白作为免疫测定试验。与任何其他对照相比,荧光MIPs与WNA的结合明显更多。因此,未来有可能开发出新的负担得起的、强大的免疫测定方法。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

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Molecular Imprinted Silica with West Nile Antibody Templates show Specific and Selective Binding in Immunoassays.

A new molecular imprinting technique was developed for molecularly imprinted polymer particles (MIPs). Particles were synthesized using organic silane chemistries by a sol-gel process, where the relative amount of active monomers was complementary matched to the relative amount of surface charges of the West Nile antibody template. Synthesized MIPs showed specific binding to affinity purified polyclonal West Nile antibodies (WNA) with a loading capacity of 80 µg/mg, while MIPs absorbed non-specific proteins at a loading capacity of 28 µg/mg. A dissociation constant of Kd=57.45 μM was measured from the binding isotherms. MIPs selectively absorbed 27 times more WNA than either albumin or immunoglobulin, while MIPs absorbed 16 times more WNA than non- imprinted particles (NIPs). Finally, fluorescently labeled MIPs were incubated in a high bind 96 well plate previously loaded with template, albumin, or immunoglobulin as an immunoassay test. Fluorescent MIPs significantly bound more to wells with WNA than any other control. Thus, the development of new affordable and robust immunoassays with MIPs would be possible in the future.

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