Abstract
Sensitive detection of small molecules using surface plasmon resonance (SPR) presents significant challenges as the antigen cannot serve as a signal generator because of its low mass; efficient binding of the target requires the binding event to be spaced from the sensor surface through a specialist linker conjugation. Competitive immunoassay of steroid hormones can be performed by conjugation through a rationally designed linker system at positions distant from existing antigenic functional groups. The binding signal from the primary antibody can then be further enhanced by sequential addition of secondary antibody or conjugated gold nanoparticles which can produce 13-fold signal enhancements through both their mass and co-operative plasmon coupling.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Homola, J. (2008) Surface plasmon resonance sensors for detection of chemical and biological species. Chem. Rev. 108, 462–93.
He, L., Musick, M. D., Nicewarner, S. R., Salinas, F. G., Benkovic, S. J., Natan, M. J., and Keating, C. D. (2000) Colloidal Au-enhanced surface plasmon resonance for ultrasensitive detection of DNA hybridization. J. Am. Chem. Soc. 122, 9071–77.
Kim, S. J., Gobi, K. V., Iwasaka, H., Tanaka, H., and Miura, N. (2007) Novel miniature SPR immunosensor equipped with all-in-one multi-microchannel sensor chip for detecting low-molecular-weight analytes. Biosens. Bioelectron. 23, 701–07.
Bieniarz, C., Husain, M., Barnes, G., King, C. A., and Welch, C. J. (1996) Extended length heterobifunctional coupling agents for protein conjugations. Bioconjug. Chem. 7, 88–95.
Wu, Y., Mitchell, J. S., Cook, C. J., and Main, L. (2002) Evaluation of progesterone-ovalbumin conjugates with different length linkers in enzyme-linked immunosorbent assay and surface plasmon resonance-based immunoassay. Steroids 67, 565–72.
Mitchell, J. S., Wu, Y., Cook, C. J., and Main, L. (2005) Sensitivity enhancement of surface plasmon resonance biosensing of small molecules. Anal. Biochem. 343, 125–35.
Otsuka, H., Nagasaki, K., and Kataoka, K. (2003) PEGylated nanoparticles for biological and pharmaceutical applications. Adv. Drug Deliv. Rev. 55, 403–19.
Mitchell, J. S., Wu, Y., Cook, C. J., and Main, L. (2006) Estrogen conjugation and antibody binding interactions in surface plasmon resonance biosensing. Steroids 71, 618–31.
Lyon, L. A., Musick, M. D., and Natan, M. J. (1998) Colloidal Au-enhanced surface plasmon resonance immunosensing. Anal. Chem. 70, 5177–83.
Sharpe, J. C., Mitchell, J. S., Lin, L., Sedoglavich, N., and Blaikie, R. (2008) Gold nanohole array substrates as immunobiosensors. Anal. Chem. 80, 2244–49.
Robertson, J. (2000) Protecting Group Chemistry. Oxford, New York.
Johnsson, B., Löfås, S., and Lindquist, G. (1991) Immobilization of proteins to a carboxymethyldextran-modified gold surface for biospecific interaction analysis in surface plasmon resonance sensors. Anal. Biochem. 198, 268–77.
Frens, G. (1973) Controlled nucleation for the regulation of the particle size in monodisperse gold suspensions. Nature. 241, 20–22.
Oliver, C. (1999) Preparation of colloidal gold. Methods Mol. Biol. 115, 327–30.
Takae, S., Akiyama, Y., Otsuka, H., Nakamura, T., Nagasaki, Y., and Kataoka, K. (2005) Ligand density effect on biorecognition by PEGylated gold nanoparticles: regulated interaction of RCA120 lectin with lactose installed to the distal end of tethered PEG strands on gold surface. Biomacromolecules 6, 818–24.
Acknowledgments
The authors would like to thank Associate Professor Lyndsay Main and Dr. Christian Cook for their advice and encouragement. This work was funded by the New Zealand Foundation for Research Science and Technology.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Mitchell, J.S., Wu, Y. (2010). Surface Plasmon Resonance Signal Enhancement for Immunoassay of Small Molecules. In: Mol, N., Fischer, M. (eds) Surface Plasmon Resonance. Methods in Molecular Biology, vol 627. Humana Press. https://doi.org/10.1007/978-1-60761-670-2_7
Download citation
DOI: https://doi.org/10.1007/978-1-60761-670-2_7
Published:
Publisher Name: Humana Press
Print ISBN: 978-1-60761-669-6
Online ISBN: 978-1-60761-670-2
eBook Packages: Springer Protocols