Energy transfer assays

Blomberg, K., Hurskainen, P. and Hemmila, I. (1999). Terbium and rhodamine as labels in a homogeneous time-resolved fluorometric energy transfer assay of the b-subunit of human chorionic gonadotropin in serum. Clin. Chem. 45, 855-61. 

Nichols, J. W. and Pagano, R. E. (1983). Resonance energy-transfer assay of protein-mediated lipid transfer between vesicles. J. Biol. Chem. 258, 5368-5371. 

Chinnayelka, S. and McShane, M. J. (2005). Microcapsule biosensors using competitive binding resonance energy transfer assays based on apoenzymes. Anal. Chem. 77, 5501-11. 

The high molar absorptivities and quantum yields of the large protein fluorophore phycoerythrin (240,000 Da) have been exploited in energy transfer assays. Phyco-erythrin has been used as both donor and acceptor, with several bound antigen molecules per phycoerythrin molecule/86,94) The usefulness of BPE is indicated in competitive assays for human IgG that use fluorescein-labeled antibody as donor to... 

Newman, M. and Josiah, S., Utilization of fluorescence polarization and time resolved fluorescence resonance energy transfer assay formats for SAR studies SRC kinase as a model system, /. Biomol. Screen., 9, 525, 2004. 

Resonance Energy Transfer Assays and Single Photon Counting Technology... 

Chinnayelka S. Microcapsule biosensors based on competitive binding and fluorescence resonance energy transfer assays. Louisiana Tech University, 2005. 

Meadows DL, Schultz JS. Design, manufacture and characterization of an optical fiber glucose affinity sensor based on an homogeneous fluorescence energy transfer assay system. Analytica Chimica Acta 1993, 280, 21-30. 

Chinnayelka S, McShane MJ. Glucose sensors based on microcapsules containing an orange/red competitive binding resonance energy transfer assay. Diabetes Technology Therapeutics 2006, 8, 269-278. 

Jeyakumar, M. and Katzenellenbogen, J.A. 2009. A dual-acceptor time-resolved Forster resonance energy transfer assay for simultaneous determination of thyroid hormone regulation of corepressor and coactivator binding to the thyroid hormone receptor. Anal. Biochem. 386, 73-78. 

Fig. 8.8. Energy transfer assays are exemplified here by a probe containing a 5 -terminal fluorescein (I) and a probe containing a 3 -end rhodamine. Upon excitation at 490 nm, fluorescein emits radiation (green fluorescence) at 520 nm. While rhodamine can be excited at 520 nm, the nonhybridized probes are not close enough for energy transfer. Probes are designed so that hybridization brings the two labels to optimal proximity. Excitation of fluorescein yields fluorescence which also serves to excite rhodamine. Rhodamine fluorescence can be specifically determined at 575 nm.

Ozers, M.S., Marks, B.D., Gowda, K., Kupcho, K.R., Ervin, K.M., De Rosier, T., Qadir, N., Eliason, H.C., Riddle, S.M. and Shekhani, M.S. (2007) The androgen receptor T877A mutant recruits LXXLL and FXXLF peptides differently than wild-type androgen receptor in a time-resolved fluorescence resonance energy transfer assay. Biochemistry, 46, 683-695. 

V. Mekler, E. Kortkhonjia, Y.W. Ebright, R.H. Ebright, Translocation of a (70) with RNA polymerase during transcription fluorescence resonance energy transfer assay for movement relative to DNA, Cell 2001, 106, 453-463. 

A study at Novartis compared hits obtained by screening 30,000 compounds for tyrosine kinase activity in three different assays a scintillation proximity assay (SPA), a homogeneous time-resolved fluorescence energy transfer assay (HT-FRET), and a fluorescence polarization (FP) assay.Library compounds were screened as mixtures of five in a well, which meant that after active wells were identified, the five constituents would be deconvoluted by screening each compound individually or using other methods to establish which of the compounds was active. " When this was done SPA, HT-FRET, and FP turned up 30, 59, and 64 hits respectively. A low degree of overlap was observed between the sets. Only four or seven compounds, depending on where the activity bars were set, would have been identified... 

Bioluminescence and Bioluminescence Resonance Energy Transfer Assays in Microfluidics... 

Bioluminescence and Bioluminescence Resonance Energy Transfer Assays in Microfluidics, Fig. 2 3D microfluidic chip for immobilized luciferase reaction [2] (With permission from MicroTas2012)... 

Bioluminescence and Bioluminescence Resonance microchamiel network, (b) Schematic of the optoelec-Energy Transfer Assays in Microfluidics, tronic detection apparatus (C/z coelenterazine 400a, DM... 

Bioluminescence and Bioluminescence Resonance Energy Transfer Assays in Mkrofluidks, Fig. 6 (a) Time-dependent BRET ratio of the control assay and the positive (27 pM) assay, (b) Dose response curve for thrombin assay a limit of detection of 19.8 pM and a sensitivity of... 

Hemmila I, Hurskainen P, Blomberg K, et al (1998) Homogeneous luminescence energy transfer assays. PCT International Application WO 98/15830... 

Fig. 1 Energy flow diagram within a europium chelate and an energy transfer assay. L ligand, A acceptor, asterisk shows excited state level...

Fluorescence Resonance Energy Transfer Assay Design... 

This method is reported to overcome some of the problems of various energy transfer assays but does still have the disadvantage that it requires two antibodies. The method has also been referred to in the literature as double antibody FIA, double-receptor FIA, indirect quenching FIA, and FIA using mixed binding reagent. 

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