摘要
本文用杂交瘤技术制备了三株抗人生长激素单克隆抗体3B10、3F3和3H3,进行了抗体性质分析。用其中一株高亲和力、高特异性的单抗3F3,建立了人血清生长激素单克隆抗体放射免疫测定法。方法学考核显示,本方法最小可测量为0.25μg/L,与hPRL和hPL的交叉结合率分别为0.07％和2.8％,批内和批间变异系数分别为4.9％和12.2％,回收率1.013±0.17,稀释试验呈线性关系。用本法测得的hGH值与多抗法相关性好(r=0.95,=1.37+0.96x),两组数据经t检验差异无显著性。用本法测定了32例正常者和8例未经治疗肢端肥大患者的上午空腹血清值,结果与临床相符。
In order to establish a highly specific and sensitive hGH-RIA, three monoclonal antibodies (MAb) against human growth hormone (hGH) were raised. The MAb 3B10, 3F3, and 3H3 were of different affinity, specificity, IgG type, litre and binding at three different combining sites of hGH. 3F3 with the highest Ka (1.7 × 1010 M-1) was used to establish a hGH-RIA. The 3F3 hybridoma ascitic fluid at a litre of 1:250 000 was used as the second antibody. The 125I-hGH was made by chloramine-T method and used as the radiolabel. The hGH was examined methodologically. The cross reactivities were 0.07% with hPRL and 2.8% with hPL; the coefficienl of varialions were 4.9% for within-assay and 12 .2% for between-assay; the minimal detectable concentration was 0.25 ±g/L; and the recovery rate was 1.013± 0.17. Dilution test showed a linear relation. The hGH levels measured by MAb-RIA were correlated well with those by PAb method in the same samples (r=0.95, y = 1.37 + 0.96 X) and no obvious difference was found between the two by t test.
The serum hGH levels of 32 healthy adults and 8 untreated acromegaly patients were measured with this method.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
1990年第3期169-171,64,共3页
Chinese Journal of Endocrinology and Metabolism
关键词
生长激素
单克隆抗体
放射免疫测定
Human growth hormone Monoclonal antibody Radioimmunoassay
