Flow injection immunoassays using solid phase immunoreactors and fluorescence detection

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.