Tandem conjugation of enzyme and antibody on silica nanoparticle for enzyme immunoassay |
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Authors: | Rongqin Ke Wei Yang Xiaohu Xia Qingge Li |
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Institution: | a Molecular Diagnostics Laboratory, Department of Biomedical Sciences and Key Laboratory of Ministry of Education for Cell Biology and Tumor Cell Engineering, School of Life Sciences, Xiamen University, Xiamen, Fujian 361005, China b Key Laboratory of Chemical Biology of Fujian, Xiamen, Fujian 361005, China |
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Abstract: | We present a new type of enzyme-antibody conjugate that simplifies the labeling procedure and increases the sensitivity of enzyme-linked immunosorbent assay (ELISA). The conjugates were prepared through layer-by-layer immobilization of enzyme and antibody on a silica nanoparticle scaffold. A maximal amount of enzyme was immobilized on the nanoparticle, followed by antibody linkage through Dextran 500. The conjugate could be easily purified from unreacted reagents by simple centrifugations. In comparison with the conventional antibody-enzyme conjugate used in ELISA, which often has one or two enzyme molecules per antibody, the new type of conjugate contained more enzyme molecules per antibody and provided a much higher signal and increased sensitivity. When used in an ELISA detection of the hepatitis B surface antigen (HBsAg), the detection limit was three times lower than that of the commercially available ELISA kit. |
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Keywords: | ELISA Silica nanoparticles Tandem conjugation Hepatitis B surface antigen |
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