Uhlich E, Weber P, Gröschel-Stewart U, Röschlau T
Horm Metab Res. 1975 Nov;7(6):501-7. doi: 10.1055/s-0028-1093713.
Antibodies for the radioimmunoassay of arginine vasopressin (AVP) described here were produced in rabbits using synthetic AVP coupled to rabbit gamma-globulin with carbodiimide. In three out of six rabbits, significant antibody titres were obtained. Using the best antisera produced, 40% of labeled AVP was bound at a final dilution of 1:50.000. After iodination of synthetic AVP with 125I using the chloramin-T method, a gel filtration on Sephadex G-25 was performed to purify the iodinated AVP. For separation of antibody bound and free hormone, a second antibody precipitation was used. There was no crossreactivity with oxytocin. AVP was extracted from plasma after ammoniumsulfate precipitation of the proteins by adsorption to Florisil. The recovery of AVP added to plasma in amounts between 5-25 pg/ml was 60 +/- 15% (n equals 6). The minimum amount of AVP detectable was 1 pg per ml plasma. The plasma level in normal adults under standard conditions was 3.4 +/- 2.2 pg/ml. This is in agreement with data recently published by other researchers. The applicability and reproducibility was further tested in measurements of samples taken hourly during the entire day under water diuresis and after hormonal stimulation of AVP.
本文所述用于精氨酸加压素(AVP)放射免疫测定的抗体是用合成的AVP通过碳二亚胺与兔γ球蛋白偶联后在兔体内产生的。六只兔中有三只获得了显著的抗体效价。使用产生的最佳抗血清,在1:50,000的最终稀释度下,40%的标记AVP被结合。用氯胺-T法将合成的AVP用125I碘化后,在Sephadex G-25上进行凝胶过滤以纯化碘化的AVP。为了分离抗体结合的和游离的激素,采用了第二抗体沉淀法。与催产素无交叉反应。通过吸附到弗罗里硅土上,在蛋白质经硫酸铵沉淀后从血浆中提取AVP。添加到血浆中浓度在5 - 25 pg/ml之间的AVP的回收率为60±15%(n = 6)。可检测到的AVP的最小量为每毫升血浆1 pg。标准条件下正常成年人的血浆水平为3.4±2.2 pg/ml。这与其他研究人员最近发表的数据一致。在水利尿期间和AVP激素刺激后,对全天每小时采集的样本进行测量,进一步测试了其适用性和可重复性。