Reduction of Non-Specific Protein Adsorption Using Poly(ethylene) Glycol (PEG) Modified Polyacrylate Hydrogels In Immunoassays for Staphylococcal Enterotoxin B Detection.
Reduction of Non-Specific Protein Adsorption Using Poly(ethylene) Glycol (PEG) Modified Polyacrylate Hydrogels In Immunoassays for Staphylococcal Enterotoxin B Detection.
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使用聚（乙烯）乙二醇（PEG）修饰的聚丙烯酯水凝胶中的非特异性蛋白吸附降低免疫测定中的葡萄球菌肠毒素B检测。
DOI：
10.3390/s90100645
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发表时间：
2009
期刊：
影响因子：
--
通讯作者：
Taitt CR
中科院分区：
文献类型：
--
作者：
Charles PT;Stubbs VR;Soto CM;Martin BD;White BJ;Taitt CR
将三种聚乙二醇单分子(聚乙二醇甲基丙烯酸酯、聚乙二醇二丙烯酸酯和聚二甲基丙烯酸酯)引入半乳糖基聚丙烯酸酯水凝胶中，测定了它们在免疫分析中减少非特异性蛋白质吸附的相对能力。形成了含胺封端官能团的高度交联水凝胶，并用于共价连接葡萄球菌肠毒素B(SEB)特异性抗体。用含有微通道的PDMS模板在聚乙二醇改性的水凝胶中创建固定抗体的图案化阵列，用于夹心免疫分析以检测SEB。将不同浓度的毒素应用于水凝胶阵列，然后使用针对这两种毒素的Cy3标记示踪抗体。示踪剂分子的荧光激光扫描共聚焦显微镜提供了检测灵敏度和由于聚乙二醇掺入而减少的非特异性结合的定性和定量测量。结果表明，聚乙二醇改性的水凝胶显著减少了非特异性蛋白的结合，SEB的检出限为1 ng/mL。荧光信号显示SEB的非特异性结合减少了10倍，而特异性结合增加了6倍。
Three PEG molecules (PEG-methacrylate, -diacrylate and -dimethacrylate) were incorporated into galactose-based polyacrylate hydrogels and their relative abilities to reduce non-specific protein adsorption in immunoassays were determined. Highly crosslinked hydrogels containing amine-terminated functionalities were formed and used to covalently attach antibodies specific for staphylococcal enterotoxin B (SEB). Patterned arrays of immobilized antibodies in the PEG-modified hydrogels were created with a PDMS template containing micro-channels for use in sandwich immunoassays to detect SEB. Different concentrations of the toxin were applied to the hydrogel arrays, followed with a Cy3-labeled tracer antibody specific for the two toxins. Fluorescence laser scanning confocal microscopy of the tracer molecules provided both qualitative and quantitative measurements on the detection sensitivity and the reduction in non-specific binding as a result of PEG incorporation. Results showed the PEG-modified hydrogel significantly reduced non-specific protein binding with a detection limit for SEB of 1 ng/mL. Fluorescence signals showed a 10-fold decrease in the non-specific binding and a 6-fold increase in specific binding of SEB.
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影响因子：
7.4
作者：
Rowe, CA;Tender, LM;Ligler, FS
通讯作者：
Ligler, FS
影响因子：
10.8
作者：
Bures, P;Huang, YB;Peppas, NA
通讯作者：
Peppas, NA
影响因子：
2.9
作者：
Angenendt, P;Glökler, J;Cahill, DJ
通讯作者：
Cahill, DJ
影响因子：
2.9
作者：
Itle, LJ;Pishko, MV
通讯作者：
Pishko, MV
影响因子：
2.2
作者：
WEDEGE, E;SVENNEBY, G
通讯作者：
SVENNEBY, G