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Fully Automated Chemiluminescence Immunoassay of Insulin Using Antibody−Protein A−Bacterial Magnetic Particle Complexes
Abstract
We report a fully automated sandwich immunoassay for the determination of human insulin using antibody-protein A-bacterial magnetic particle (BMP) complexes and an alkaline phosphatase-conjugated secondary antibody. BMPs bearing protein A-MagA inserted on the external surface of the membrane were prepared in the Magnetospirillum sp. AMB-1 transconjugant for a protein A-magA fusion gene. MagA protein was used as an anchor to attach protein A onto the membrane. Protein A-BMP complexes harvested from transconjugant AMB-1 were subsequently complexed with anti-human insulin antibodies by specific binding between the Z domain of protein A and the Fc component of IgG to form the antibody-protein A-BMP complexes. The complexes were quite monodisperse after the binding of the antibody. The BMPs' monodispersity resulted in high signal and low noise in the immunoassay. The luminescence intensity ((kilocounts/s)/microg of antibody) from antibody-protein A-BMP complexes after immunoreaction was higher than that from BMPs chemically conjugated to an antibody. This was explained by a difference in dispersion. The fully automated sandwich immunoassay system using antibody-protein A-BMP complexes made possible precise assays of human insulin in serum.
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Citations
Cited by
Liu, J.-F., Zhao, Z.-S., & Jiang, G.-B. (2008) · Environmental Science & Technology · doi:10.1021/es800924c · Cited by 1,084
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Yi, D. K., Selvan, S. T., Lee, S. S., Papaefthymiou, G. C., Kundaliya, D., & Ying, J. Y. (2005) · Journal of the American Chemical Society · doi:10.1021/ja0428863 · Cited by 851
Nanoparticle Architectures Templated by SiO2/Fe2O3 Nanocomposites
Yi, D. K., Lee, S. S., Papaefthymiou, G. C., & Ying, J. Y. (2006) · Chemistry of Materials · doi:10.1021/cm0512979 · Cited by 381
Biotechnological applications of bioluminescence and chemiluminescence
RODA, A. (2004) · Trends in Biotechnology · doi:10.1016/j.tibtech.2004.03.011 · Cited by 357
Zhao, X., Shi, Y., Cai, Y., & Mou, S. (2008) · Environmental Science & Technology · doi:10.1021/es071817w · Cited by 295
Metadata
- Authors
- Tanaka, T., & Matsunaga, T.
- Venue
- Analytical Chemistry
- Publisher
- American Chemical Society (ACS)
- Published
- 2000-06-24
- Volume
- 72
- Issue
- 15
- Pages
- 3518-3522
- ISSN
- 0003-2700 (ISSN-L)
- 1520-6882
- OpenAlex
- W2011715535
- Cited by
- 250
- References
- 0
- FWCI
- 2.70
- Type
- journal article
- Language
- en
- Open access
- closed
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- Tanaka, T., & Matsunaga, T. (2000). Fully Automated Chemiluminescence Immunoassay of Insulin Using Antibody−Protein A−Bacterial Magnetic Particle Complexes. Analytical Chemistry. https://doi.org/10.1021/ac9912505
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