用荧光探针标记肽以掺入可降解聚合物中。

Labelling peptides with fluorescent probes for incorporation into degradable polymers.

作者信息

Brunner A, Minamitake Y, Göpferich A

机构信息

University of Erlangen-Nürnberg, Germany.

出版信息

Eur J Pharm Biopharm. 1998 May;45(3):265-73. doi: 10.1016/s0939-6411(98)00009-5.

DOI:10.1016/s0939-6411(98)00009-5

PMID:9653631

Abstract

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.

摘要

两种肽，即心钠素（ANP）和鲑鱼降钙素（sCT），与一种荧光胺反应性探针5-（和6-）羧基四甲基罗丹明琥珀酰亚胺酯（5-（6）-TAMRA-SE）进行了偶联。标记反应通过高效液相色谱（HPLC）进行监测，发现2小时后反应完成。标记后的肽通过凝胶过滤色谱法进行纯化，并通过[1H]核磁共振（NMR）、紫外可见（UV/VIS）光谱和荧光光谱进行表征。核磁共振光谱证实了染料与肽的偶联。在紫外可见光谱中记录到500至600纳米之间的两个吸收最大值。发现荧光光谱依赖于pH值，这使得能够测量水溶液中的pH值。使用双乳液技术将标记后的肽包封到聚乳酸（PLA）微球中。探针的连接使得能够确定肽在聚合物中的位置。