Endo Y, Miyai K, Hata N, Ichihara K
Anal Biochem. 1985 Jan;144(1):41-6. doi: 10.1016/0003-2697(85)90081-8.
Immunoglobulin G fractions prepared from conventional rabbit anti-thyroxine (T4) antisera were fractionated by agarose gel isoelectric focusing (IEF) in the range of pH 3 to 10, and by chromatofocusing using a Fast Protein Liquid Chromatography (FPLC) system. The clonotype antibodies were recovered from the fractions and subjected to Scatchard plot analysis. The highest affinity constants of the initial antibody (shown in parentheses) and those of the antibodies recovered were IEF, 1.8 X 10(9) to 8.3 X 10(9) M-1 (2.2 X 10(9) M-1); FPLC, 2.4 X 10(9) to 6.0 X 10(9) M-1 (2.5 X 10(9) M-1). A sensitive radioimmunoassay of T4 was achieved with the isolated high-affinity anti-T4 antibody. The minimum detectable concentration of T4 was 6.3 X 10(-15) to 1.5 X 10(-14) mol/tube, which was three to five times lower than detectable with the initial antibodies.
从传统兔抗甲状腺素（T4）抗血清制备的免疫球蛋白G组分，通过在pH 3至10范围内的琼脂糖凝胶等电聚焦（IEF）以及使用快速蛋白质液相色谱（FPLC）系统的色谱聚焦进行分级分离。从这些组分中回收克隆型抗体，并进行Scatchard作图分析。初始抗体（括号内显示）和回收抗体的最高亲和常数分别为：IEF，1.8×10⁹至8.3×10⁹ M⁻¹（2.2×10⁹ M⁻¹）；FPLC，2.4×10⁹至6.0×10⁹ M⁻¹（2.5×10⁹ M⁻¹）。用分离出的高亲和力抗T4抗体实现了对T4的灵敏放射免疫测定。T4的最低可检测浓度为6.3×10⁻¹⁵至1.5×10⁻¹⁴ mol/管，这比初始抗体的可检测浓度低三至五倍。
