Labelling peptides with fluorescent probes for incorporation into degradable polymers.
Two peptides, atrial natriuretic peptide (ANP) and salmon calcitonin (sCT) were conjugated with a fluorescent, amine-reactive probe 5-(and 6-)carboxytetramethylrhodamine,-succinimidylester (5-(6)-TAMRA-SE). The labelling reaction was followed by HPLC and found to be complete after 2 h. The labelled peptides were purified by gel filtration chromatography and characterised by [1H]NMR, UV/VIS and fluorescence spectroscopy. NMR-spectra confirmed the conjugation of dye to the peptides. Two absorption maxima between 500 and 600 nm were recorded in the UV/VIS-spectra. The fluorescence spectra were found to be pH-dependent, which allowed the measurement of pH in aqueous solution. The labelled peptides were encapsulated into poly(lactic acid) (PLA) microspheres using a double emulsion technique. Probe attachment permitted location of the peptides in the polymer.
Brunner
A
A
University of Erlangen-Nürnberg, Germany.
Minamitake
Y
Y
Göpferich
A
A
D009320:Atrial Natriuretic Factor / Q000737:chemistry; D002116:Calcitonin / Q000737:chemistry; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004655:Emulsions; D005456:Fluorescent Dyes / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D008863:Microspheres; D010455:Peptides / Q000737:chemistry*; D011108:Polymers; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D004655:Emulsions; D005456:Fluorescent Dyes; D010455:Peptides; D011108:Polymers; C028815:salmon calcitonin; D009320:Atrial Natriuretic Factor; D002116:Calcitonin