Abstract
The Z-domains of protein A was expressed as a fusion protein at the outer membrane of E.coli by using the autodisplay technology. Because of the specific affinity towards the Fc region of immunoglobulins (IgG’s), the Z-domains have been used for the orientation control of antibodies in order to improve the sensitivity of immunoassays. In this work, the E.coli with autodispalyed Z-domains was immobilized to the SPR biosensor by the charge interaction. The surface modification was carried out by covalent layering of the poly-L-lysine with amino groups to the parylene-H film with formyl groups. And then, the negatively charged E.coli cells were immobilized by charge interaction with the positivley charged of poly-L-lysine. The effectiveness of this layer for the immobilization of E.coli was estimated by counting the number of E.coli cells in comparison with the bare gold surface and the poly-L-lysine coated gold surface. For the test of feasibility of the immobilized E.coli cells to SPR biosensor, the stability of immobilized E.coli cells was estimated by treatment of salt solutions at the known concentrations to the immobilized E.coli cells which were bound through the charge interaction. From this test, the E.coli cells immobilized to the parylene-H film with poly-L-lysine coating were determined to be stable at the salt concentration of human serum. Then, the applicability of the immobilized E.coli cells with autodisplayed Z-domains was demonstrated by detection of C-reactive protein (CRP). The effect of orientation control with autodisplayed Z-domains was estimated by comparing the sensivities by immobilization through the physical adsorption and charge interaction to poly-L-lysine coated layer.
Similar content being viewed by others
References
Benz, I. & Schmidt, M.A. AIDA-1, the adhesion involved in diffuse adherence of the diarrhoeagenic Escherichia coli strain 2787 (O1126:H27), is synthesized via a precursor molecule. Mol. Microbiol. 6, 1539–1546 (1992).
Mauer, J., Jose, J. & Meyer, T.F. Autodisplay: onecomponent system for efficient surface display and release of soluble recombinant proteins from Escherichia coli. J. Bacteriol. 179, 794–804 (1997).
Jose, J. et al. Escherichia coli with autodisplayed Zdomain of protein A for signal amplification of SPR biosensor. Biosens Bioelectron 24, 1324–1329 (2009).
Jose, J., Park, M. & Pyun, J.C. Highly sensitive immunoassay based on E.coli with autodisplayed Z-domain. Anal. Chim. Acta 667, 113–118 (2010).
Yoo, G., Park, M., Lee, E.H., Jose, J. & Pyun, J.C. Immobilization of E.coli with autodisplyaed Z-domains to a surface-modified microplate for immunoassay. Anal. Chim. Acta 707, 142–147 (2011).
Lee, J.K. et al. Electrochemical ELISA based on E.coli with autodisplayed Z-domains. Sensors and Actuators B in press (2012).
Jose, J., Park, M. & Pyun, J.C. E.coli outer membrane with autodisplayed Z-domain as a molecular recognition layer of SPR biosensor. Biosensors and Bioelectronics 25, 1225–1228 (2010).
Park, M., Jose, J. & Pyun, J.C. Hyper sensitive immunoassay by using E.coli outer membrane with autodisplayed Z-domains. Enzyme and Microbial Technology 46, 309–314 (2010).
Park, M., Jose, J. & Pyun, J.C. SPR biosensor by using E.coli outer membrane with autodisplayed Z-domain. Sensors and Actuators B 154, 82–88 (2011).
Homora, J., Yee, S.S. & Myszka, D. Ch4. Surface plasmon resonance biosensors., In: Ligler FS and Taitt CR, editors. Optical Biosensors: Today and Tomorrow (2nd eds.). Elsevier BV, p. 185–210 (2008).
Jeon, B.J., Kim, M.H. & Pyun, J.C. Application of a functionalized parylene film as a linker layer of SPR biosensor. Sensors and Actuators B 154, 89–95 (2011).
Ko, H. et al. One step immobilization of peptides and proteins by using modified parylene with formyl groups. Biosensors and Bioelectronics 30, 56–60 (2011).
Hiroshi, N. Outer membrane. In: Neidhardt FC, Umbarger HE, editors. Escherichia coli and Salmonella typhimurium: cellular and molecular biology. Washington: ASM Press, p. 29–47 (1996).
Bugg, T.D.H., Braddick, D., Dowson, C.G. & Roper, D.I. Bacterial cell wall assembly: still an attractive antibacterial target. Trends Biotechnol. 29, 167–173 (2011).
Koch, S., Woias, P., Meixner, L.K., Drost, S. & Wolf, H. Protein detection with a novel ISFET-based zeta potential analyzer. Biosens. Bioelectron. 14, 413–421 (1999).
Chang, M.H., Dosev, D. & Kennedy, I.M. Zeta-potential analyses using micro-electrical field flow fractionation with fluorescent nanoparticles. Sens. Actuators B. 124, 172–178 (2007).
Ledue, T.B. & Rifai, N. Preanalytic and analytic sources of variations in C-reactive protein measurement: implications for cardiovascular disease risk assessment. Clin. Chem. 49, 1258–1271 (2003).
Kimberly, M.M. et al. Standardization of immunoassays for measurement of high sensitivity C-reactive protein. Phase I: evaluation of secondary reference materials. Clin. Chem. 49, 611–616 (2003).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Lee, EH., Yoo, G., Jose, J. et al. SPR biosensor based on immobilized E.coli cells with autodisplayed Z-domains. BioChip J 6, 221–228 (2012). https://doi.org/10.1007/s13206-012-6304-3
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s13206-012-6304-3