Savastano S, Tommaselli A P, Valentino R, Carlino M, Selleri A, Randazzo G, Benvenga S, Lombardi G
Cattedra di Endocrinologia, Facoltà di Medicina e Chirurgia, Università Federico II, Napoli, Italy.
J Endocrinol Invest. 1995 Jan;18(1):9-16. doi: 10.1007/BF03349690.
The aim of this study was to evaluate the feasibility of a chromatographic method to easily detect circulating anti-thyroid hormone autoantibodies (THBA), to calculate their affinity constant and the total thyroid hormone (TH) levels in presence of THBA. This method was applied to sera from 4 subject with suspected THBA and 20 controls (10 normal subjects and 10 patients with thyroid dysfunctions). After a short incubation with 125I-T3 or T4 tracer solution containing 8-anilino-1-naphthalenesulfonic acid as inhibitor of the binding of TH to plasma proteins without affecting THBA binding, the samples were chromatographied on prepacked Sephadex LH 20 columns and eluted with TRIS buffer to separate free TH from THBA-bound TH. Samples were considered THBA positive when radioactivity values in TRIS eluates were higher than in controls. THBA-bound TH were subsequently eluted with methanol and used to calculate the total TH present in patients with THBA. After a validation test using two standardized methods, we propose this method to quickly detect TH-BA in samples with suspected spuriously high values of free and total TH.
本研究的目的是评估一种色谱方法的可行性,该方法可轻松检测循环中的抗甲状腺激素自身抗体(THBA),计算其亲和常数以及在存在THBA的情况下总甲状腺激素(TH)水平。该方法应用于4名疑似THBA患者的血清和20名对照(10名正常受试者和10名甲状腺功能障碍患者)。在与含有8-苯胺基-1-萘磺酸的125I-T3或T4示踪剂溶液短暂孵育后,该溶液作为TH与血浆蛋白结合的抑制剂但不影响THBA结合,然后将样品在预装的Sephadex LH 20柱上进行色谱分析,并用TRIS缓冲液洗脱以分离游离TH和与THBA结合的TH。当TRIS洗脱液中的放射性值高于对照时,样品被认为是THBA阳性。随后用甲醇洗脱与THBA结合的TH,并用于计算患有THBA患者体内的总TH。在使用两种标准化方法进行验证测试后,我们提出该方法可快速检测游离TH和总TH值疑似异常高的样品中的THBA。