Single chain fragment variable recombinant antibody functionalized gold nanoparticles for a highly sensitive colorimetric immunoassay.
Single chain fragment variable recombinant antibody functionalized gold nanoparticles for a highly sensitive colorimetric immunoassay.
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DOI：
10.1016/j.bios.2009.02.016
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发表时间：
2009-05-15
影响因子：
12.6
通讯作者：
Zeng X
中科院分区：
文献类型：
--
作者：
Liu Y;Liu Y;Mernaugh RL;Zeng X
在本报告中，连接scFv VH和VL结构域的肽接头经遗传修饰以含有不同的氨基酸（即半胱氨酸（scFv-cys）或组氨酸（scFv-his）），以使scFv能够吸附或自组装到金纳米颗粒（NP）上。将scFv-cys稳定的金纳米颗粒用于开发高灵敏度的比色免疫传感器。通过紫外-可见光谱、透射电子显微镜（TEM）和红外光谱（FT-IR）对scFv-cys稳定的金纳米颗粒进行表征，加入兔IgG抗原后，scFv-cys稳定的金纳米颗粒溶液由于金纳米颗粒的聚集而出现明显的可见颜色变化，由深红色变为浅紫色。基于scFv-cys稳定的金纳米颗粒的比色聚集，免疫传感器具有较高的灵敏度，检测限为1.7 nM，并具有良好的特异性。该免疫传感器的良好性能归因于scFv的小尺寸以及scFv与金纳米颗粒之间的共价连接，其改善了更好的取向并提高了探针密度。基于功能化单链抗体稳定的金纳米颗粒的比色免疫测定具有快速、简单和特异性的优点，代表了蛋白质分析和临床诊断的一种有前途的方法。
In this report, the peptide linker connecting scFv VH and VL domains were genetically modified to contain different amino acids (i.e. cysteine (scFv-cys) or histidines ( scFv-his)) to enable the scFv to adsorb or self-assemble onto the gold nanoparticles (NPs). The scFv-cys stabilized gold NPs were used to develop a highly sensitive colorimetric immunosensor. The scFv-cys stabilized gold NPs were characterized by UV-vis spectra, transmission electron microscope (TEM) and FT-IR. After adding the antigen rabbit IgG, the solution of scFv-cys stabilized gold NPs shows obvious visible color change from deep red to light purple due to the aggregation of the gold nanoparticles. Based on the colorimetric aggregation of scFv-cys stabilized gold NPs, the immunosensor exhibits high sensitivity with detection limit of 1.7 nM and good specificity. The good properties of the colorimetric aggregation immunosensor would be attributed to the small size of scFv and the covalent link between the scFv and gold NPs that improve the better orientation and enhance the probe density. With the advantages of speed, simplicity and specificity, the colorimetric immunoassay based on the functionalized scFv stabilized gold NPs represents a promising approach for protein analysis and clinical diagnostics.
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DOI：
10.1073/pnas.0504917102
发表时间：
2005-10-11
影响因子：
11.1
作者：
Backmann, N;Zahnd, C;Gerber, C
通讯作者：
Gerber, C
影响因子：
7.4
作者：
Shen, ZH;Mernaugh, RL;Zeng, XQ
通讯作者：
Zeng, XQ
影响因子：
2.9
作者：
Zhang, CX;Zhang, Y;Lu, ZH
通讯作者：
Lu, ZH
影响因子：
15
作者：
Liu, JW;Lu, Y
通讯作者：
Lu, Y
影响因子：
7.4
作者：
Huang, CC;Huang, YF;Chang, HT
通讯作者：
Chang, HT