Rajashekharrao B, Kumar A, Velumani A, Shah D H
Radiation Medicine Centre, Bhabha Atomic Research Centre, Bombay.
Indian J Med Res. 1992 Dec;96:350-5.
A two-site immunoradiometric assay (IRMA) for estimation of serum thyroglobulin (tg) is optimized using partially purified rabbit anti-tg antibodies (ammonium sulphate precipitated) as a solid phase antibody and affinity purified 125I labeled anti-tg antibodies as a tracer. The IRMA correlated well (r = 0.93, n = 200, P < 0.001) with conventional radioimmunoassay (RIA). The inter- and intra-assay coefficient of variation were comparable for both methods. The IRMA method gave a superior sensitivity and working range (2 ng/ml, 3-1000 ng/ml) as compared to that of RIA method (6 ng/ml, 9-500 ng/ml). Using IRMA the total incubation time was curtailed to 4 h as compared to 90 h required for conventional RIA. The shelf-life of tracer antibody used in IRMA was much longer (3 months) as compared to tracer tg (3-4 wk) used in RIA. The optimized IRMA for quantitation of serum tg has superior assay characteristics as compared to RIA.
采用部分纯化的兔抗甲状腺球蛋白(Tg)抗体(硫酸铵沉淀法)作为固相抗体,亲和纯化的¹²⁵I标记抗Tg抗体作为示踪剂,对用于估算血清甲状腺球蛋白(Tg)的双位点免疫放射分析(IRMA)进行了优化。该IRMA与传统放射免疫分析(RIA)具有良好的相关性(r = 0.93,n = 200,P < 0.001)。两种方法的批间和批内变异系数相当。与RIA方法(6 ng/ml,9 - 500 ng/ml)相比,IRMA方法具有更高的灵敏度和工作范围(2 ng/ml,3 - 1000 ng/ml)。使用IRMA时,总孵育时间缩短至4小时,而传统RIA需要90小时。与RIA中使用的示踪剂Tg(3 - 4周)相比,IRMA中使用的示踪剂抗体的保质期要长得多(3个月)。用于定量血清Tg的优化IRMA与RIA相比具有更好的分析特性。