Miller J J, Schultz G S, Levy R S
Int J Pept Protein Res. 1984 Aug;24(2):112-22. doi: 10.1111/j.1399-3011.1984.tb00935.x.
A simple, rapid method is described for the purification of radioiodinated peptides for use in radioimmuno- and in radioreceptor assays. Iodinated reaction mixtures are applied directly onto Sep-Pak disposable, reversed phase cartridges equilibrated with phosphate buffer. Unreacted 125-iodide and other non-peptide reaction components are eluted with buffer. The peptide fraction is then eluted with 70% buffer: 30% acetonitrile. The peptide fraction is further purified by reversed phase high pressure liquid chromatography to separate the native peptide and the mono- and diiodo-derivatives. In this study the method is used to prepare 125-iodide-labeled monoiodo-leucine enkephalin and monoiodo-angiotensin II, which are free of the parent peptides and diiodo-derivatives and are of maximum obtainable specific radioactivity. The usefulness of these labeled peptides in radioimmuno- and radioreceptor assays is demonstrated by their binding to specific antibodies and receptors, respectively.
本文描述了一种简单、快速的方法，用于纯化放射性碘化肽，以用于放射免疫分析和放射受体分析。将碘化反应混合物直接应用于用磷酸盐缓冲液平衡的Sep-Pak一次性反相柱上。用缓冲液洗脱未反应的125碘化物和其他非肽反应成分。然后用70%缓冲液：30%乙腈洗脱肽部分。通过反相高压液相色谱进一步纯化肽部分，以分离天然肽以及单碘和二碘衍生物。在本研究中，该方法用于制备125碘标记的单碘亮氨酸脑啡肽和单碘血管紧张素II，它们不含亲本肽和二碘衍生物，且具有最大可获得的比放射性。这些标记肽分别与特异性抗体和受体结合，证明了它们在放射免疫分析和放射受体分析中的实用性。
