Peptides fluorescence detection

Baars, M.J. and Patonay, G., Ultrasensitive detection of closely related angiotensin I peptides using capillary electrophoresis with near-infrared laser-induced fluorescence detection, Anal. Chem. 71, 667, 1999. 

Mao, Y., Zhang, X.M. (2003). Comprehensive two-dimensional separation system hy coupling capillary reverse-phase liquid chromatography to capillary isoelectric focusing for peptide and protein mapping with laser-induced fluorescence detection. Electrophoresis 24, 3289-3295. 

Solinova V, Kasicka V, Koval D et al (2004) Analysis of synthetic derivatives of peptide hormones by capillary zone electrophoresis and micellar electrokinetic chromatography with ultraviolet-absorption and laser-induced fluorescence detection. J Chromatogr B 808 75-82... 

Zhang H, Le Potier I, Smadja C et al (2006) Fluorescent detection of peptides and amino acids for capillary electrophoresis via on-line derivatization with 4-fluoro-7-nitro-2,l, 3-benzoxadiazole. Anal Bioanal Chem 386 1387-1394... 

Huang, Y., Duan, J., Chen, H., Chen, M., and Chen, G. (2005). Separation and determination of carnosine-related peptides using capillary electrophoresis with laser-induced fluorescence detection. Electrophoresis 26, 593-599. 

In one report, DTT was added in the run buffer for the CE separation of fluorescently labeled peptides with fluorescent detection. What is the purpose of adding DTT [1039] (2 marks)... 

In a protein-binding assay with fluorescence detection a microarray of biotin, HPYPP-peptide and WSHHPQFEK-peptide was screened against streptavidin-Cy3 and avidin-Cy5. By following the same principle an anti-human insulin monoclonal antibody was also screened against a set of different peptides. 

C. Detection of N-terminal Variants Using Peptide Mapping and Fluorescence Detection... 

Wehr, C. T. (1991). Post column reaction systems for fluorescence detection of polypeptides. In High-Performance Liquid Chromatography of Peptides and Proteins Separation, Analysis, and Conformation (C. T. Mant and R. S. Hodges, eds.), pp. 579-586. CRC Press, Boca Raton, FL. 

Nishino, N., Makinose, Y., and Fujimoto, T. 1992. 2-Aminobenzoyl-peptide-2,4-dinitroanilinoethylamides facile fluorescent detection system for sequence specific proteases. Chem. Lett. 1, 77-80. 

Although the peptide aminomethylcoumarin (AMC) substrate approach has been widely used for measuring the activities of many proteases, a disadvantage of this approach arises from the ultraviolet excitation and emission wavelengths required for detecting free AMC where there is a possibility for fluorescence interference. RllO-labeled substrates are less encumbered by interference due to a spectral red shift, but many compounds in small molecule chemical libraries exhibit fluorescent properties that may interfere with assays based on fluorescence detection (Simenov et al. 2008). 

Recently, a small molecule fluorophore phosphosensor technology referred as Pro-Q Diamond dye has been developed to detect and quantitate phosphorylated amino acids within peptides and proteins in microarrays. ° In addition to binding assays, fluorescence detection methods have also been developed for functional assays. For example, microarrays of quenched fluorescent substrates can be used to detect protease or esterase activities in the analytes. In this method, quenched fluorescent substrates are prepared by coupling the peptide substrate to coumarin, a fluorescent dye. These peptide substrates are then spotted onto the solid support... 

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