posted on 2018-05-11, 15:49authored byJiu-Chun Guo
Fluorescence immunoassays have been established for a number of years as
valuable methods of analysis in clinical chemistry and other fields, being sensitive,
safe, easy to use and available in a variety formats. Those in common use are
normally single analyte assays. But in many cases (e.g. forensic drug screening,
therapeutic drug monitoring, screening for cancer markers, monitoring of thyroid
function, or the analysis of environmental pollutants) dual- or multi-analyte assays
would be much more valuable, with the advantages of increased information
content, savings in time and costs, and the elimination of some sources of sampling
variance. Amongst all the labels used in immunoassays, only fluorescent groups
offer realistic prospects of practicable multi-analyte assays.
This project has investigated single-, dual- and multi-analyte fluorescence
immunoassays using several spectroscopic and software methods to resolve
multicomponent fluorescence emission or synchronous spectra. The assays have
been based on flow injection analysis methodology, with solid phase reactors to
effect the separation of antibody-bound and unbound labelled analytes. The use of
solid phase reactors incorporating thiophiIic gels to bind antibodies has also been
investigated: these stationary phases have the advantage that bound antibodies can
be eluted by changes of ionic strength, rather than changes of pH. This allows the
use of a much wider range of fluorescence labels, clearly important in multi-analyte
assays, and it has thus proved possible to develop successful dual and triple analyte
assays, with results that compare well with other independent methods.
This work is made available according to the conditions of the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0) licence. Full details of this licence are available at: https://creativecommons.org/licenses/by-nc-nd/4.0/
Publication date
1998
Notes
A Doctoral Thesis. Submitted in partial fulfilment of the requirements for the award of Doctor of Philosophy at Loughborough University.