Abstract: Bacterial outer membrane proteins, along with a filling lipid molecule can be modified to form stable self-assembled monolayers on gold. The transmembrane domain of Escherichia coli outer membrane protein A has been engineered to create a scaffold protein to which functional motifs can be fused. In earlier work we described the assembly and structure of an antibody-binding array where the Z domain of Staphylococcus aureus protein A was fused to the scaffold protein. Whilst the binding of rabbit polyclonal immunoglobulin G (IgG) to the array is very strong, mouse monoclonal IgG dissociates from the array easily. This is a problem since many immunodiagnostic tests rely upon the use of mouse monoclonal antibodies. Here we describe a strategy to develop an antibody-binding array that will bind mouse monoclonal IgG with lowered dissociation from the array. A novel protein consisting of the scaffold protein fused to two pairs of Z domains separated by a long flexible linker was manufactured. Using surface plasmon resonance the self-assembly of the new protein on gold and the improved binding of mouse monoclonal IgG were demonstrated.
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Le Brun, A.P.; Shah, D.S.H.; Athey, D.; Holt, S.A.; Lakey, J.H. Self-Assembly of Protein Monolayers Engineered for Improved Monoclonal Immunoglobulin G Binding. Int. J. Mol. Sci. 2011, 12, 5157-5167.
Le Brun AP, Shah DSH, Athey D, Holt SA, Lakey JH. Self-Assembly of Protein Monolayers Engineered for Improved Monoclonal Immunoglobulin G Binding. International Journal of Molecular Sciences. 2011; 12(8):5157-5167.
Le Brun, Anton P.; Shah, Deepan S. H.; Athey, Dale; Holt, Stephen A.; Lakey, Jeremy H. 2011. "Self-Assembly of Protein Monolayers Engineered for Improved Monoclonal Immunoglobulin G Binding." Int. J. Mol. Sci. 12, no. 8: 5157-5167.