Baylis P H, Heath D A
Clin Endocrinol (Oxf). 1977 Aug;7(2):91-102. doi: 10.1111/j.1365-2265.1977.tb01300.x.
A radioimmunoassay for AVP capable of measuring human plasma AVP is described. Iodination was performed by the chloramine T method and purified by chromatography on Sephadex G-25. Specific activity of 125I-AVP was 1710 +/- 155 Ci/mmol. Antiserum of high affinity (Keq = 2.7 X 10(11) 1/mol) has been raised in rabbits, which shows slight cross-reactivity with LVP and negligible reactivity with oxytocin. The aqueous assay is capable of detecting 0.4 fmol of AVP/tube and it is highly reproducible. A F lorisil extraction technique is described in detail and gives recovery of 70% of synthetic AVP added to plasma over a wide physiological range. The lowest detectable concentration of plasma AVP is 0.3 pmol/l. The method has been validated by studying changes in plasma AVP concentration following overnight dehydration (plasma AVP =3.46 +/- 1.89 (SD) pmol/l), and water loading (plasma AVP = 1.54 +/- 0.59 pmol/l), P less than 0.005, in normal subjects. A highly significant positive correlation has been found between plasma AVP and plasma osmolality (r =+0.75). Plasma AVP concentration has also been determined in patients with DI and the syndrome of inappropriate ADH secretion. No effect was found on the level of plasma AVP in normally hydrated volunteers undergoing postural change but levels rose following strenuous exercise from basal concentrations of 1.57 +/- 0.59 pmol/l to 4.77 +/- 3.43 pmol/l, P less than 0.01.
本文描述了一种能够检测人血浆中抗利尿激素(AVP)的放射免疫分析方法。采用氯胺T法进行碘化,并通过Sephadex G - 25柱色谱法进行纯化。125I - AVP的比活度为1710±155 Ci/mmol。已在兔体内制备出高亲和力(平衡常数Keq = 2.7×10¹¹ 1/mol)的抗血清,该抗血清与赖氨酸加压素(LVP)有轻微交叉反应,与催产素的反应可忽略不计。该水溶液分析方法能够检测出每管0.4 fmol的AVP,且具有高度可重复性。详细描述了一种弗罗里硅土萃取技术,在很宽的生理范围内,该技术对添加到血浆中的合成AVP的回收率为70%。血浆AVP的最低可检测浓度为0.3 pmol/l。通过研究正常受试者过夜脱水后(血浆AVP = 3.46±1.89(标准差)pmol/l)和水负荷后(血浆AVP = 1.54±0.59 pmol/l)血浆AVP浓度的变化,验证了该方法,P<0.005。血浆AVP与血浆渗透压之间存在高度显著的正相关(r = +0.75)。还测定了尿崩症(DI)和抗利尿激素分泌不当综合征患者的血浆AVP浓度。体位改变对正常水合状态志愿者的血浆AVP水平无影响,但剧烈运动后,血浆AVP水平从基础浓度1.57±0.59 pmol/l升至4.77±3.43 pmol/l,P<0.01。