Single chain fragment variable recombinant antibody as a template for Fc sensors.

Single chain fragment variable recombinant antibody as a template for Fc sensors.
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DOI:
10.1021/ac102087w
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发表时间:
2011-01-15
影响因子:
7.4
通讯作者:
Zeng, Xiangqun
Zeng, Xiangqun
中科院分区:
化学1区
文献类型:
--
作者:
Yan, Heping;Shen, Zhihong;Mernaugh, Ray;Zeng, Xiangqun

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研制了一种用于检测细胞表面Fc受体的无标记免疫传感器,并用石英晶体微平衡(QCM)传感器对其进行了表征。利用单链片段变量(single chain fragment variable, scFv)重组抗体的特点和抗体的多效性,将重组抗体固定在预形成的功能化自组装单层(SAMs)模板表面。单分子ScFv可以与任何兔IgG的CH1区结合,形成一个高度定向的IgG层,其Fc部分指向溶液相。这就产生了一种高度定向的Fc传感器,可用于研究免疫球蛋白Fc部分与细胞表面Fc受体(FcR)结合的热力学和动力学,这是免疫系统的一个重要领域。利用Fc传感器研究了金黄色葡萄球菌(S. aureus)与巨噬细胞与Fc受体的结合。用ELISA法对相同样品的细胞表面Fc受体进行平行表征。
A label free immunosensor for detection of Fc receptors expressed on cell surface was developed and characterized by Quartz Crystal Microbalance (QCM) transducer. Taking advantage of the characteristics of single chain fragment variable (scFv) recombinant antibody and the multivalency of an antibody, the engineered recombinant scFv was immobilized onto pre-formed functionalized self-assembled monolayers (SAMs) template surface. The monomeric ScFv can bind with the CH1 region of any rabbit IgG to form a highly oriented IgG layer with its Fc portion pointing toward solution phase. This results in a highly oriented Fc sensor that can be used to study the thermodynamics and kinetics of binding between Fc portion of immunoglobulin and cell surface Fc receptor (FcR), an important area of the immune system. The Fc sensor was used to study the binding between Staphylococcus aureus (S. aureus) and Macrophage with Fc receptor. Parallel characterization of cell surface Fc receptors in the same samples by ELISA was also performed.
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