PubMed · 10351982
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D W Chan, L J Sokoll. 1999. Prostate-specific antigen: advances and challenges.. https://pubmed.ncbi.nlm.nih.gov/10351982/
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The immunoassay, based on specific recognition of an antigen by its antibody, has garnered widespread use in clinical analysis as well as application in such areas as food industry and environmental monitoring. Fluoroimmunoassays (FIAs) are especially attractive due to the inherent sensitivity of fluorescence spectroscopy and the availability of a wide range of commercial antibodies and fluorescent labels. In current form, however, FIAs can be cumbersome, multistep procedures and often lack versatility when there is interest in measuring many different target antigens. This report is proof of a concept paper introducing a new FIA approach, Magnetically-Assisted Transport Evanescent Field Fluoroimmunoassays (MATEFFs), which seeks to preserve the advantages of current approaches to FIAs while attempting to address some of the drawbacks. MATEFFs utilize magnetic microspheres as solid supports for the fluoroimmunoassay with direct detection of bound analyte within the sample mixture effected by selectively driving the functionalized beads to a prism surface using an external magnet. An evanescent wave is generated by total internal reflection of a laser beam at the optical interface between the prism and sample and serves to excite the fluorescent species magnetically delivered into the localized field. This technique eliminates wash steps without compromising sensitivity, all the while minimizing interference from fluorescing species present in the sample matrix. Preliminary optimization studies assessing the impact of background interfering agents, incident angle, magnetic field direction, laser power density via focusing, and bead concentration on MATEFFs performance characteristics are discussed herein along with a detailed description of the experimental platform. Utilizing a model sandwich assay system with biotinylated anti-IgG as the capture antibody, rabbit IgG as the antigen, and anti-IgG-R-phycoerythrin as the reporter antibody, we demonstrate a linear dynamic range of 3 orders of magnitude, physiologically relevant detection limits of low nanograms per milliliter, and RSD values of less than 5%.
A novel class of 1-(isoquinolin-5-yl)-4-arylamino-phthalazines is described as inhibitors of vascular endothelial growth factor receptor II (VEGFR-2). Many compounds display VEGFR-2 inhibitory activity with an IC(50) as low as 0.017 microM in an HTRF enzymatic assay. The compounds also inhibit VEGFR-1, a related tyrosine kinase.
OBJECTIVE: To prepare time-resolved fluoroimmunoassay (TRFIA) kit for detecting human alpha-fetoprotein (hAFP). METHODS: Sandwich TRFIA kit for hAFP detection was developed using monoclonal antibody of anti-AFP. RESULTS: AFP-TRFIA kit was capable of detecting AFP within the range of 1-1 000 U/ml with sensitivity of 0.17 U/ml and without cross-reactivity with CEA, CA12-5, CA15-3, or CA19-9. The intra- and inter-assay coefficients of variation were (3.3-5.9)% and (3.7-6.5)%, respectively. The prepared AFP-TRFIA reagent could withstand preservation at 4 degrees celsius; for 1 year and at 37 degrees celsius; for 7 days with the cutoff value of 12 U/ml in healthy subjects (n=426). The correlation coefficient of the detection results between this kit and commercially available AFP kit (Wallac OY, Finland) for 60 blood samples was 0.995. CONCLUSION: The prepared TRFIA kit for hAFP detection meets the demand of clinical application with good sensitivity, precision, specificity, stability and accuracy.