Surf.'1ce enhanced resonance Raman scattering (SERRS) is an extremely sensitive detection technique, comparable to or, in some circumstances, better than fluorescence. Thanks to the molecularly specific vibrational signals, SERRS enables the identification of a specific label in situ in the presence of other materials or other labels. SERRS is produced when an analyte containing a chromophore is bound onto a.suitably roughened metal surface. The signals are recorded using Raman spectroscopy and the excitation frequency must match or be close to both the plasmon resonance frequency of the metal and an electronic transition of the chromophore. This thesis reports the development of an antibody assay using SERRS detection using colloidal silver particles as the substrate. An appropriate SERRS labelling chemistry was developed, by synthesising new SERRS labels using dyes, a polymer dye and SERRS active beads. They were conjugated to an antibody. SERRS of the labelled antibody is intense and gives specitic peaks which identify the label. Under the conditions used, antibodies were detected with a better sensitivity by SERRS detection (2.79 x 10-13 mol.dm-1 than by fluorescence (3.46 x 10-10 mol.dm·3 ). A sandwich assay was developed for two targets, brain natriuretic peptide and holotranscobalamin: The first target was used to assess the potential of several bioassay formats. The second target was added later to determine the potential for a multiple target assay. Optimisation of a complete SERRS assay using magnetic microparticles was carried out. Quantitative results were obtained with anyone run but there was variability between runs. A final investigation focused on trying to understand the source of the variability.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:486539 |
| Date | January 2007 |
| Creators | Sabatte, Gwenola |
| Publisher | University of Strathclyde |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
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