The use of flow injection analysis with fluorescence detection was evaluated
using the host-guest phenomenon between the cyclodexnins and DL-Iysine and Lserine.
Auorescence enhancement, kinetic and equilibrium studies were recorded and
the effect of pH and time on fluorescence were also observed.
Rhodamine isothiocyanate was conjugated to insulin. Insulin and dye were
mixed in different ratios, and the dye : insulin ratio was detennined for each
conjugate. These conjugates were checked for immunoreactivity. Insulin-biotin and
antibody-iminobiotin conjugates were also prepared. Insulin : biotin ratio was also
determined. An insulin-biotin .. avidin-Texas Red complex was also prepared. Each was
checked for immunoreactivity.
Protein G-agarose, protein A-controlled pore glass(CPG), streptavidin-agarose,
and avidin D-agarose-biotin-antibody solid phase immunoreactors were used in flow
injection immunoassay of insulin. In these immunoassays, antibody, insulin and
labelled insulin were incubated in vitro and then injected onto the immunoreactor. A
binding buffer carried the sample through the immunoreactor and a fluorescent
detector. An acidic buffer then eluted the components of the sample bound to the
immunoreactor, which were then measured. An assay range for insulin was developed
in each solid phase assay.
A Doctoral Thesis. Submitted in partial fulfilment of the requirements for the award of Doctor of Philosophy of Loughborough University