A rapid and sensitive chemiluminescent immunoassay of total thyroxin with DMAE x NHS-Labeled.

A novel chemiluminescence immunoassay (CLIA) of total thyroxin (TT4) with the acridinium ester of DMAE x NHS [2',6'-dimethyl-4'-(n-succinimidyloxycarbonyl) phenyl-10-methyl-acridinium-9-carboxylate methosulfate] labeled has been developed. In our method, microwells were coated with anti-T4 monoclonal antibody (McAb) and DMAE x NHS synthesized by our laboratory was conjugated with streptavidin (SA). T4-BSA was conjugated with biotin-N-hydroxysuccinimide (B x NHS), T4-BSA- B x NHS and the T4 in the standard or sample competitively react with anti-T4 McAb.

Sensitive time-resolved fluoroimmunoassay for simultaneous detection of serum thyroid-stimulating hormone and total thyroxin with Eu and Sm as labels.

Based on a novel cocoating strategy and dissociation enhancement lanthanide fluorescence immunoassay technique, a sensitive time-resolved fluoroimmunoassay (TRFIA) has been developed for simultaneous quantification of human serum thyroid-stimulating hormone (TSH) and thyroxin (T4) in a one-and-the-same assay procedure. The new cocoating strategy for preparing highly active surface anti-TSH and anti-T4 monoclonal antibodies (McAbs) was performed by a three-step protocol. Namely, anti-TSH McAb at high concentration (10 micro g/ml) and extensively biotinylated bovine serum albumin (BSA) at low concentration (0.

Synthesis of a highly fluorescent beta-diketone-europium chelate and its utility in time-resolved fluoroimmunoassay of serum total thyroxine.

A new highly fluorescent beta-diketone-europium chelate was synthesized and employed as a tracer to develop a time-resolved fluoroimmunoassay (TRFIA) for detection of serum total thyroxine (T4). The tetradentate beta-diketone chelator, 1,10-bis(thiophene-2'-yl)-4,4,5,5,6,6,7,7-octafluorodecane-1,3,8,10-tetraone (BTOT), was structurally composed of two units of thenoyltrifluoroacetone (TTA) derivatives but expressed fluorescence that was greatly enhanced, as compared to the original TTA molecules, in the presence of excess amount of Eu3+. The luminescence properties of the europium chelate of BTOT werestudied in aqueous solution.

Time-resolved immunofluorometry of serum hTSH with enhanced sensitivity.

Sensitive TSH immunoassays offer a clear advance in discriminating the TSH concentrations in serum between hyperthyroid and euthyroid individuals; they have been proposed as the best single screening test for thyroid disorders. We have developed a highly sensitive serum TSH TRFIA based on DELFIA technology. Three monoclonal antibodies (McAbs) directed against different epitopes of the TSH molecule were involved in this assay, of which, one McAb was used to coat clear microwells, and the other two were biotinylated' for signal generation after being bound by the europium labeled streptavidin.