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Pappalysin-1,又称妊娠相关血浆蛋白A,由PAPPA基因编码,其主要底物是胰岛素样生长因子结合蛋白。Pappalysin-1也被用于唐氏综合征的筛查测试。该基因编码的是一种分泌型金属蛋白酶,可裂解胰岛素样生长因子结合蛋白(IGFBPs)。PAPPA的蛋白分解功能在胶原蛋白结合时被激活。它被认为参与了局部增殖过程,如伤口愈合和骨骼重塑。该蛋白的低血浆水平被认为是非整倍体胎儿(染色体数目异常的胎儿)怀孕的生化标志。例如,低PAPPA可能在产前唐氏综合征筛查中常见。低水平也可能预测胎盘问题,导致不良并发症,如宫内生长受限、子痫前期、胎盘早剥、早产或胎儿死亡。 本试剂盒采用竞争法酶联免疫吸附试验(ELISA)。往预先包被有人妊娠相关血浆蛋白A(PAPP-A)抗原的微孔中,依次加入样本、标准品、Biotin标记的抗体,HRP酶结合物,中间经过温育和洗涤,用底物TMB显色,TMB在过氧化物酶(HRP)的催化下转化成蓝色,并在酸的作用下转化成最终的黄色。颜色的深浅和样本中的人妊娠相关血浆蛋白A(PAPP-A)呈负相关。用酶标仪在450nm波长下测定吸光度(OD值),计算样本浓度。
This kit employs the competitive enzyme-linked immunosorbent assay (ELISA) principle. Microplate wells are pre-coated with Human PAPP-A antigen. The sample, standards, a biotin-labeled antibody, and an HRP enzyme conjugate are then added sequentially to the wells. The procedure includes incubation and wash steps between each addition. TMB substrate is used for color development. In the presence of the peroxidase (HRP) enzyme, TMB is catalyzed to produce a blue color, which is then converted into a final stable yellow color upon the addition of an acid stop solution. The intensity of the resulting yellow color is inversely proportional to the concentration of Human PAPP-A in the sample. The optical density (OD) is measured at a wavelength of 450 nm using a microplate reader, and the concentration of the target analyte in the sample is calculated based on the standard curve.