Self-assembly of protein monolayers engineered for improved monoclonal immunoglobulin G binding

Thumbnail Image
Journal Title
Journal ISSN
Volume Title
MDPI Publishing
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. © The Authors - This is an open access article distributed under the Creative Commons Attribution License
Membrane proteins, Gold, Immunoglobulins, Resonance, Surfaces, Molecules
Le Brun, A.P., Shah, D.S.H., Athey, D., Holt, S.A., Lakey, J.H. (2011). Self-assembly of protein monolayers engineered for improved monoclonal immunoglobulin G binding. International Journal of Molecular Sciences, 12(8), 5157-5167. doi:10.3390/ijms12085157