Choi M J, Song E Y, Chung T W
Bioanalysis & Biotransformation Research Center, Korea Institute of Science & Technology, Seoul, Korea.
Arch Pharm Res. 1998 Jun;21(3):231-5. doi: 10.1007/BF02975280.
A sensitive enzyme immunoassay for serum TSH has been developed utilizing the tight binding between biotin and avidin, and three layered protein polystyrene beads as solid phase. To increase binding capacity of TSH and sensitivity of the assay, the polystyrene beads were coated sequentially with mouse immunoglobulin as first layer, rabbit antimouse immunoglobulin as second layer and monoclonal anti-TSH as third layer. A serum sample was incubated simultaneously with a monoclonal anti-TSH immobilized polystyrene beads and a second monoclonal anti-TSH covalently attached to biotin. After washing, the antibody bound serum TSH-anti-TSH-biotin complex is reacted with horseradish peroxidase (HRP)-labeled avidin. Following a second wash, the bound HRP activity was measured colorimetrically. Reproducible results were obtained within 4 hours for serum TSH in the range between 0 microIU/ml and 50 microIU/ml with detection limit of 0.1 microIU per test.
利用生物素与抗生物素蛋白之间的紧密结合以及三层蛋白质聚苯乙烯珠作为固相，开发了一种用于血清促甲状腺激素（TSH）的灵敏酶免疫测定法。为了提高TSH的结合能力和测定的灵敏度，聚苯乙烯珠依次用小鼠免疫球蛋白作为第一层、兔抗小鼠免疫球蛋白作为第二层和单克隆抗TSH作为第三层进行包被。将一份血清样本与固定有单克隆抗TSH的聚苯乙烯珠以及共价连接有生物素的第二种单克隆抗TSH同时孵育。洗涤后，将抗体结合的血清TSH-抗TSH-生物素复合物与辣根过氧化物酶（HRP）标记的抗生物素蛋白反应。第二次洗涤后，通过比色法测量结合的HRP活性。对于0微国际单位/毫升至50微国际单位/毫升范围内的血清TSH，在4小时内可获得可重复的结果，每次检测的检测限为0.1微国际单位。
