一种使用反相高效液相色谱法制备用于受体研究的125I标记人生长激素的快速方法。

A rapid method for the preparation of 125I-labelled human growth hormone for receptor studies, using reverse-phase high performance liquid chromatography.

作者信息

Ilondo M M, Dehart I, De Meyts P

出版信息

Biochem Biophys Res Commun. 1986 Jan 29;134(2):671-7. doi: 10.1016/s0006-291x(86)80472-7.

Abstract

Human growth hormone was labelled with 125 Iodine by the stoichiometric modification of the chloramine-T method to a specific activity of 50-80 microCi/microgram, and the iodinated mixture was purified by reverse-phase high performance liquid chromatography using a C18 column (SynChropak RP-P) and a linear gradient. Compared with the usual Sephadex G-100 chromatography, HPLC gave a much better separation, with a higher yield and a considerably reduced analysis time (30 min vs 5 h). The [125I]-labelled preparation had normal binding to IM-9 lymphocyte receptors. The maximum bindability of the HPLC-purified preparation approximated 90%, which is the best value so far reported for human growth hormone. It is concluded that HPLC is a fast, convenient and reproducible method for obtaining an improved [125I]-labelled human growth hormone for receptor studies.

摘要

采用氯胺-T 法的化学计量修饰法，将人生长激素用 125碘标记，使其比活度达到 50 - 80 微居里/微克，然后使用 C18 柱（SynChropak RP-P）和线性梯度，通过反相高效液相色谱法对碘化混合物进行纯化。与常规的葡聚糖 G - 100 色谱法相比，高效液相色谱法分离效果更好，产率更高，分析时间大幅缩短（30 分钟对 5 小时）。[125I]标记制剂与 IM - 9 淋巴细胞受体具有正常结合能力。经高效液相色谱法纯化的制剂的最大结合能力约为 90%，这是迄今为止报道的人生长激素的最佳值。结论是，高效液相色谱法是一种快速、便捷且可重复的方法，用于获得改进的[125I]标记人生长激素以用于受体研究。