Chow S N, Ho-Yuen B, Lee C Y
J Appl Biochem. 1985 Apr;7(2):114-21.
Monoclonal antibodies to human luteinizing hormone (hLH) were generated by a modified hybridoma technique. Out of forty hybrid cell lines that were shown to secrete antibodies reacting with hLH, LH35 and LH40 were further characterized biochemically and immunologically. LH35 was found to secrete immunoglobulin G1 antibody specific to the alpha-subunit of LH, whereas that of LH40 was beta-subunit specific. Association constants between the antibodies and LH were determined to be 2 X 10(8) and 1 X 10(9) M-1, respectively, by using competitive radioimmunoassay and Scatchard plots. Monoclonal antibodies from LH35 and LH40 were purified from the respective ascites fluids by ammonium sulfate fractionations and DEAE ion-exchange chromatography. The purified alpha-subunit-specific antibody of LH35 was immobilized on polystyrene balls (6 mm in diameter), whereas purified LH40 antibody was conjugated with horseradish peroxidase or labeled with iodine-125. Solid-phase radio- and enzyme immunoassays were designed to measure relatively low concentrations of LH (2-100 mIU/ml). The LH surge during the midcycles of women with normal menstrual cycles could easily be detected from daily urine or serum specimens by a 1-h assay procedure. It is proposed that this new LH immunoassay procedure can be routinely used for predicting ovulation of women with normal menstrual cycles.
采用改良的杂交瘤技术制备了抗人促黄体生成素（hLH）的单克隆抗体。在显示能分泌与hLH反应的抗体的40个杂交细胞系中，对LH35和LH40进行了进一步的生化和免疫学特性鉴定。发现LH35分泌针对LHα亚基的免疫球蛋白G1抗体，而LH40分泌的抗体则针对β亚基。通过竞争性放射免疫分析和Scatchard作图法，测定抗体与LH之间的结合常数分别为2×10⁸和1×10⁹M⁻¹。通过硫酸铵分级沉淀和DEAE离子交换色谱法，从各自的腹水中纯化了来自LH35和LH40的单克隆抗体。将纯化的LH35的α亚基特异性抗体固定在聚苯乙烯球（直径6毫米）上，而纯化的LH40抗体则与辣根过氧化物酶偶联或用碘-125标记。设计了固相放射免疫分析和酶免疫分析方法来测定相对低浓度的LH（2 - 100 mIU/ml）。通过1小时的检测程序，能够轻松地从正常月经周期女性的每日尿液或血清标本中检测到月经周期中期的LH峰。建议这种新的LH免疫分析方法可常规用于预测正常月经周期女性的排卵情况。
