Sato H, Mochizuki H, Tomita Y, Kanamori T
Fuji Central Research Laboratory, Mochida Pharmaceutical Co., Ltd., Tokyo, Japan.
Clin Biochem. 1996 Dec;29(6):509-13. doi: 10.1016/s0009-9120(96)00098-7.
To develop a highly sensitive chemiluminescent immunoassay (CLIA) that measures the serum estradiol (E2) levels in postmenopausal women.
The previously developed competitive CLIA for E2 consisted of an immobilized antigen and a labeled antibody with an N-functionalized acridinium ester that was modified to enhance its sensitivity. The modifications were changing the immobilized antigen from E2 to its analogues with its expected effect on hapten heterology and selecting optimal incubation conditions.
The hapten heterology in which estriol was used as the immobilized antigen instead of E2 enhanced the sensitivity of the CLIA about 3-fold. A low incubation temperature and a long incubation time also effectively increased the sensitivity of the CLIA. The combination of these modifications enhanced sensitivity about 10-fold. The proposed CLIA with a 16 pmol/L detection limit, was about 5-fold more sensitive than commercially available immunoassay kits for E2.
The proposed CLIA is sensitive enough to measure serum E2 levels in postmenopausal women.
开发一种高灵敏度化学发光免疫分析法（CLIA），用于测定绝经后女性血清雌二醇（E2）水平。
先前开发的用于E2的竞争性CLIA由固定化抗原和标记抗体组成，标记抗体带有经修饰以增强其灵敏度的N-功能化吖啶酯。修饰包括将固定化抗原从E2改为其类似物，观察其对半抗原异质性的预期影响，并选择最佳孵育条件。
使用雌三醇而非E2作为固定化抗原的半抗原异质性使CLIA的灵敏度提高了约3倍。低孵育温度和长孵育时间也有效提高了CLIA的灵敏度。这些修饰的组合使灵敏度提高了约10倍。所提出的CLIA检测限为16 pmol/L，比市售E2免疫分析试剂盒灵敏约5倍。
所提出的CLIA灵敏度足以测定绝经后女性血清E2水平。
