Pollet D E, Nouwen E J, Schelstraete J B, Renard J, Van de Voorde A, De Broe M E
Clin Chem. 1985 Jan;31(1):41-5.
In this enzyme-antigen immunoassay for human placental alkaline phosphatase (hPLAP; EC 3.1.3.1.) in serum and tissue extracts, polyclonal rabbit antiserum to mouse IgG2b is adsorbed to the wells of a microtiter plate, its excess binding sites are blocked, then it is incubated with murine monoclonal anti-hPLAP and mixed with serially diluted standard or sample antigen. The amount of antigen bound is determined by measuring its enzymic activity. The standard curve is linear for hPLAP concentrations of 0.2 to 1 U/L. The mean within-assay CV was 3.8% (SD 0.9%) for a serum sample and 6.1% (SD 3.0%) for a tissue extract. The respective mean between-assay CVs were: 6.7% (SD 2.0%), and 7.0% (SD 2.0%). Serum hPLAP concentrations, determined in four different dilutions, had a CV of 5.5%. We evaluated the method by standard additions and by comparing dilution curves for purified hPLAP, hPLAP in serum, and hPLAP in tissue extracts. The upper limit of activity in normal subjects was 0.1 U/L for serum samples, and 1.0 mU/g wet weight of tissue for tissue extracts. hPLAP activity was increased in 9.8% of all cancer patients, and in 40% of ovarian cancer patients. Almost half of the tumor biopsies were positive for hPLAP activity, and 94% of the biopsies from ovarian neoplasia had an increased activity of this isoenzyme. Of the nonmalignant tissues examined, normal lung tissue had the highest hPLAP activity.
在这种用于检测血清和组织提取物中人胎盘碱性磷酸酶（hPLAP；EC 3.1.3.1.）的酶 - 抗原免疫测定中，将兔抗小鼠IgG2b多克隆抗血清吸附到微量滴定板孔中，封闭其多余的结合位点，然后与鼠抗hPLAP单克隆抗体一起孵育，并与系列稀释的标准抗原或样品抗原混合。通过测量其酶活性来确定结合的抗原量。对于hPLAP浓度为0.2至1 U/L，标准曲线呈线性。血清样品的批内平均变异系数（CV）为3.8%（标准差0.9%），组织提取物的批内平均CV为6.1%（标准差3.0%）。各自的批间平均CV分别为：6.7%（标准差2.0%）和7.0%（标准差2.0%）。在四种不同稀释度下测定的血清hPLAP浓度的CV为5.5%。我们通过标准加入法以及比较纯化hPLAP、血清中的hPLAP和组织提取物中的hPLAP的稀释曲线来评估该方法。正常受试者血清样品的活性上限为0.1 U/L，组织提取物为每克湿重组织1.0 mU。在所有癌症患者中，9.8%的患者hPLAP活性升高，卵巢癌患者中40%的患者hPLAP活性升高。几乎一半的肿瘤活检hPLAP活性呈阳性，94%的卵巢肿瘤活检该同工酶活性升高。在所检查的非恶性组织中，正常肺组织的hPLAP活性最高。
