CLINICAL EVALUATION OF A TWO‐SITE IMMUNORADIOMETRIC ASSAY FOR ADRENOCORTICOTROPIC IN UNEXTRACTED HUMAN PLASMA USING MONOCLONAL ANTIBODIES
CLINICAL EVALUATION OF A TWO‐SITE IMMUNORADIOMETRIC ASSAY FOR ADRENOCORTICOTROPIC IN UNEXTRACTED HUMAN PLASMA USING MONOCLONAL ANTIBODIES
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使用单克隆抗体对未提取人血浆中促肾上腺皮质激素进行双位点免疫放射测定的临床评价
DOI：
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发表时间：
1987
期刊：
影响因子：
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通讯作者：
J. G. Ratcliffe
中科院分区：
文献类型：
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作者：
Anne White;H. Smith;M. Hoadley;S. Dobson;J. G. Ratcliffe
我们建立了一种灵敏的人血浆中完整ACTH及其前体、前阿片黑素皮质素和22 kDa多肽的双部位免疫放射分析(IRMA)方法。这项检测使用了两种单抗。针对ACTH 10-18的抗体1A12被放射性标记，而针对C-末端区域(ACTH 24-39)的抗体2A3与Sephacryl S300偶联用于固相。样品与标记抗体孵育18h，然后与固相抗体孵育2 h。分离采用蔗糖分层技术。以含10%马血清的稀释液中的人垂体ACTH1-39(编码74/555)为标准品，其灵敏度为3.5±0.8 ng/L(n=7)。本方法批内平均变异系数为5.9%，批间平均变异系数为6.7%，在5,000 ng/L浓度范围内小于10%，加入地塞米松抑制的人血浆中ACTH1-39的平均回收率为109%，内源性ACTH的稀释度与标准ACTH无差别。正常人血浆促肾上腺皮质激素水平0730h为30 ng/L，1630h为15 ng/l。促肾上腺皮质激素浓度在口服美替拉酮后8~10.5h(口服2 g，2300h)，胰岛素诱导低血糖后30~60min，地塞米松4 ng/l，8h(口服1.5g，2300h)之间。在一系列病理条件下，ACTH浓度准确地反映了垂体-肾上腺轴的紊乱。内源性ACTH免疫活性在体外22℃下，全血至少1小时，血浆至少4小时内稳定。结论：双部位免疫放射分析法检测未提取血浆中ACTH为临床提供了一种可靠的检测方法。
We have developed a sensitive two‐site immunoradiometric assay (IRMA) for intact ACTH and its precursors, pro‐opiomelanocortin and 22 kDa peptide in unextracted human plasma. The assay uses two monoclonal antibodies. Antibody 1A12, specific for ACTH 10–18, is radiolabelled and antibody 2A3 specific for the C‐terminal region (ACTH 24–39), is coupled to Sephacryl S300 for the solid‐phase. Samples are incubated for 18 h with labelled antibody followed by 2 h with solid‐phase antibody. Separation employs the sucrose layering technique. Using human pituitary ACTH 1–39 (code 74/555) in diluent containing 10% horse serum to standardize the assay, the sensitivity (upper 99% confidence limit of zero standard) is 3.5 ± 0.8 ng/l (n= 7). The mean coefficient of variation is 5.9% within‐assay and 6.7% between‐assay and is less than 10% between 22 and >5000 ng/l. Mean recovery of ACTH 1–39 added to dexamethasone‐suppressed human plasma is 109% and endogenous ACTH behaves indistinguishably from standard ACTH on dilution. In normal subjects, mean plasma ACTH levels are 30 ng/l at 0730 h, and 15 ng/1 at 1630 h at rest. ACTH concentrations are between 60 and 330 ng/l, 8–10.5 h after metyrapone (2 g orally at 2300 h), between 140 and 320 ng/1, 30–60 min after insulin‐induced hypoglycaemia, and < 4 ng/1, 8 h after dexamethasone (1.5 mg orally at 2300 h). In a range of pathological conditions ACTH concentrations accurately reflect the disorders of the pituitary‐adrenal axis. Endogenous ACTH immunoactivity is stable in vitro at 22°C for at least 1 h in whole blood and at least 4 h in plasma. It is concluded that this two‐site IRMA for ACTH in unextracted plasma offers a reliable assay for clinical purposes.
未提取的人血浆中促肾上腺皮质激素的快速放射免疫测定。
影响因子：
9.3
作者：
Nicholson,WE;Davis,DR;Sherrell,BJ;Orth,DN
通讯作者：
Orth,DN