Paul S, Wood K, Said S I
Peptides. 1984 Nov-Dec;5(6):1085-7. doi: 10.1016/0196-9781(84)90175-x.
VIP was labeled with sodium 125iodide, and 125I-VIP was purified by reverse-phase high performance liquid chromatography. Optimal separations of 125I-VIP and unlabeled VIP were obtained using two C18-Novapak columns in series and a gradient of acetonitrile in triethylamine phosphate for elution. The specific activity of the 125I-VIP was 1.99 +/- 0.21 Ci/mumole, approaching the maximum specific activity of monoiodinated VIP (2.26 Ci/mumole). Radioimmunoassay and radioreceptorassay for VIP were more sensitive (2.6-fold, and 2.5-fold, respectively) using 125I-VIP purified by HPLC compared to 125I-VIP obtained from an open-end cellulose column. These results demonstrate the advantage of preparing purified 125I-VIP by HPLC for the accurate assay of VIP and VIP-receptors in tissues and biological fluids.
血管活性肠肽（VIP）用125碘化钠进行标记，然后通过反相高效液相色谱法对125I-VIP进行纯化。使用两根串联的C18-Novapak柱以及乙腈在磷酸三乙胺中的梯度洗脱，可实现125I-VIP与未标记VIP的最佳分离。125I-VIP的比活为1.99±0.21 Ci/微摩尔，接近单碘化VIP的最大比活（2.26 Ci/微摩尔）。与通过开放式纤维素柱获得的125I-VIP相比，使用经高效液相色谱法纯化的125I-VIP进行VIP的放射免疫测定和放射受体测定更为灵敏（分别提高了2.6倍和2.5倍）。这些结果证明了通过高效液相色谱法制备纯化的125I-VIP对于准确测定组织和生物体液中的VIP及VIP受体的优势。
