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Int. J. Mol. Sci. 2011, 12(8), 5157-5167; doi:10.3390/ijms12085157
Article

Self-Assembly of Protein Monolayers Engineered for Improved Monoclonal Immunoglobulin G Binding

1,†
, 2
, 2
, 3,†
 and 1,*
Received: 4 July 2011 / Revised: 4 July 2011 / Accepted: 10 August 2011 / Published: 15 August 2011
(This article belongs to the Special Issue Molecular Self-Assembly 2011)
View Full-Text   |   Download PDF [467 KB, 19 June 2014; original version 19 June 2014]   |   Browse Figures

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.
Keywords: membrane protein; gold; immunoglobulin; self-assembled monolayer; surface plasmon resonance membrane protein; gold; immunoglobulin; self-assembled monolayer; surface plasmon resonance
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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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.

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