Lewis J R, Morley J S, Venn R F
Pain Research Institute, Pain Relief Foundation, Liverpool, UK.
J Chromatogr. 1993 May 19;615(1):37-45. doi: 10.1016/0378-4347(93)80288-f.
A method is described for the determination of sub-picomole amounts of Lys-Lys-Gly-Glu [the C-terminal tetrapeptide sequence of human beta-endorphin, referred to as melanotropin potentiating factor (MPF)], a putative neurotrophic agent. Attempts to raise antibodies to the peptide were not successful and we have therefore developed a method based on the fluorescence of its 9-fluorenylmethyl chloroformate (FMOC) derivative which provides a sensitivity comparable to that of radioimmunoassay. Standard solutions, cerebrospinal fluid or central nervous tissue extracts are first treated with FMOC-Cl. The resulting mixture of FMOC-peptides is then subjected to high-performance liquid chromatography (HPLC) and quantified using a fluorescence monitor. By this procedure, MPF and related peptides can be analysed from one sample in a single HPLC run. The method was also applied to determine the rate of release into a phosphate-buffered saline medium of a metabolically stable analogue of MPF from a slow-release formulation of the compound.
本文描述了一种用于测定痕量皮摩尔水平的赖氨酸-赖氨酸-甘氨酸-谷氨酸[人β-内啡肽的C端四肽序列，称为促黑素增强因子（MPF）]的方法，MPF是一种假定的神经营养因子。制备针对该肽的抗体的尝试未成功，因此我们开发了一种基于其9-芴甲基氯甲酸酯（FMOC）衍生物荧光的方法，该方法提供了与放射免疫分析相当的灵敏度。标准溶液、脑脊液或中枢神经组织提取物首先用FMOC-Cl处理。然后将所得的FMOC-肽混合物进行高效液相色谱（HPLC）分析，并用荧光监测器进行定量。通过该程序，MPF和相关肽可以在一次HPLC运行中从一个样品中进行分析。该方法还用于测定化合物缓释制剂中MPF代谢稳定类似物释放到磷酸盐缓冲盐溶液介质中的速率。
