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- W875865332 abstract "The use of fluorescein and rhodamine as donor andacceptor fluorescent labels in the development of energytransfer immunoassay (ETIA) methods was evaluated by developingan assay for human serum albumin. The sensitivity of the assaywas found to depend on (i) the degrees of fluorophore labellingof antibody and antigen, (ii) concentrations of labelledantibody and antigen, (iii) the fluorimeter spectral bandwidth,and (iv) whether the donor (fluorescein) was conjugated to theantigen or the antibody. These results, including those relatingto the stability of the labelled immune reactants on storage,lead to the conclusion that fluorescein and rhodamine are farfrom ideal as donor and acceptor. Nevertheless, the applicationof the assay to the analysis of test serum samples gave resultsthat compared favourably with those obtained by electroimmunoassay.Other potential donor and acceptor fluorescent labelswere also investigated, viz. Ca) fluorescamine and fluorescein,(b) MDPF and fluorescein, (c) dansyl chloride and rhodamine,(d) quinacrine and fluorescein, and (e) quinacrine and rhodamine.Of these, only (a) and (b) were found to be suitable donor-acceptorpairs, and they were applied to the development of a number ofassays for both low and high molecular-weight analytes. Comparative studies of fluorescamine and MDPF as donor fluorescentlabels together with fluorescein as the acceptor label wereperformed by developing immunoassay methods for the determinationof human serum transferrin in four serum samples including ablood sample from the victim of a road traffic accident. Resultsobtained were generally in good agreement with those found bythe radial immunodiffusion method. An ETIA developed ·fornortriptyline and related tricyclic antidepressants was capableof detecting nanomolar concentrations of the drugs in puresolution and in spiked sera. Other ETIA's developed include asandwich assay for the quantitation of human immunoglobulin A,and a direct assay for the determination of human immunoglobulin G.The Fluram as well as the MDPF enhancement phenomenawere also studied in detail and applied successfully to thedevelopment of fluorescence enhancement immunoassays fornortriptyline, human serum transferrin and immunoglobulin G.Finally, the automation of an energy transferimmunoassay was successfully performed by using the principles ofstopped-flow injection analysis with merging zones." @default.
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- W875865332 date "1980-01-01" @default.
- W875865332 modified "2023-09-23" @default.
- W875865332 title "The development of energy transfer immunoassay methods" @default.
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