Fully automated immunoassay for detection of prostate-specific antigen using nano-magnetic beads and micro-polystyrene bead composites, 'Beads on Beads' |
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Authors: | Matsunaga Tadashi Maeda Yoshiaki Yoshino Tomoko Takeyama Haruko Takahashi Masaaki Ginya Harumi Aasahina Junko Tajima Hideji |
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Institution: | a Department of Biotechnology, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan b Precision System Science Co. Ltd., 88 Kamihongo, Matsudo, Chiba 271-0064, Japan |
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Abstract: | Magnetic beads have served as a conventional bioassay platform in biotechnology. In this study, a fully automated immunoassay was performed using novel nano- and microbead-composites constructed by assembling nano-magnetic beads onto polystyrene microbeads, designated ‘Beads on Beads’. Nano-sized bacterial magnetic particles (BacMPs) displaying the immunoglobulin G (IgG)-binding domain of protein A (ZZ domain) were used for the construction of ‘Beads on Beads’ via the interaction of biotin-streptavidin. The efficient assembly of ‘Beads on Beads’ was performed by gradual addition of biotin-labeled BacMPs onto streptavidin-coated polystyrene microbeads. Approximately 2000 BacMPs were uniformly assembled on a single microbead without aggregation. The constructed ‘Beads on Beads’ were magnetized and separated from the suspension by using an automated magnetic separation system with a higher efficiency than BacMPs alone. Furthermore, fully automated detection of prostate-specific antigens was performed with the detection limit of 1.48 ng mL−1. From this preliminary assay, it can be seen that ‘Beads on Beads’ could be a powerful tool in the development of high-throughput, fully automated multiplexed bioassays. |
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Keywords: | Fully automated immunoassay Prostate-specific antigen Bacterial magnetic particle Polystyrene microbeads |
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