Bionanocapsule-based enzyme–antibody conjugates for enzyme-linked immunosorbent assay

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• 作者：Masumi Iijima ; Takashi Matsuzaki ; Hiroyasu Kadoya ; Satoko Hatahira ; Shingo Hiramatsu ; Giman Jung ; Katsuyuki Tanizawa ; Shun’ ; ichi Kuroda
• 关键词：ELISA ; IgG ; Protein A ; Nanoparticles ; Oriented immobilization
• 刊名：Analytical Biochemistry
• 出版年：2010
• 出版时间：15 January 2010
• 年：2010
• 卷：396
• 期：2
• 页码：257-261
• 全文大小：351 K

文摘

Macromolecules that can assemble a large number of enzyme and antibody molecules have been used frequently for improvement of sensitivities in enzyme-linked immunosorbent assays (ELISAs). We generated bionanocapsules (BNCs) of approximately 30 nm displaying immunoglobulin G (IgG) Fc-binding ZZ domains derived from Staphylococcus aureus protein A (designated as ZZ-BNC). In the conventional ELISA using primary antibody and horseradish peroxidase-labeled secondary antibody for detecting antigen on the solid phase, ZZ-BNCs in the aqueous phase gave an approximately 10-fold higher signal. In Western blot analysis, the mixture of ZZ-BNCs with secondary antibody gave an approximately 50-fold higher signal than that without ZZ-BNCs. These results suggest that a large number of secondary antibody molecules are immobilized on the surface of ZZ-BNCs and attached to antigen, leading to the significant enhancement of sensitivity. In combination with the avidin–biotin complex system, biotinylated ZZ-BNCs showed more significant signal enhancement in ELISA and Western blot analysis. Thus, ZZ-BNC is expected to increase the performance of various conventional immunoassays.