Design and Application of a Microarray for Fluorescence and Surface Plasmon Resonance Imaging Analysis of Peptide-Antibody Interactions
Authors: Cherif, Boutheina; Villiers, Christian L.; Paranhos-Baccalà, Glaucia; Calemczuk, Roberto; Marche, Patrice N.; Livache, Thierry; Villiers, Marie-Bernadette
Source: Journal of Biomedical Nanotechnology, Volume 2, Number 1, April 2006 , pp. 29-35(7)
Publisher: American Scientific Publishers
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Abstract:
In the past few years, protein and peptide microarrays have shown a great potential in fundamental research, medical diagnostics and drug discovery. We have developed a new method based on covalent immobilization of peptides on a microarray gold surface for detecting antibodies by either fluorescence or surface plasmon resonance imaging (SPRi). The fluorescence-based revelation approach allows for an indirect analysis of protein-antibody binding via labeled molecules, whereas the SPRi-based assay is a real-time and label-free detection system suitable for the direct analysis of biological interactions. We illustrate our peptide chip's efficiency in specifically binding serum antibodies and compare the fluorescence and SPRi approaches to detect peptide-antibody interactions. Moreover, we used a monoclonal antibody against the Hepatitis C virus nucleocapsid protein to provide evidence that our system is a powerful tool to identify which epitope is recognized by a given antibody. This novel technology therefore provides a promising tool for both antibody screening and epitope mapping.Keywords: PEPTIDE CHIP; SPR IMAGING; ANTIBODY; PYRROLE; HEPATITIS C VIRUS
Document Type: Research article
DOI: 10.1166/jbn.2006.005
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