Itkonen O, Koivunen E, Hurme M, Alfthan H, Schröder T, Stenman U H
Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Finland.
J Lab Clin Med. 1990 Jun;115(6):712-8.
We have developed sensitive time-resolved immunofluorometric assays for the two trypsinogen isoenzymes, trypsinogen-1 and trypsinogen-2, which also are called cationic and anionic trypsinogen, respectively. The assays use monoclonal antibodies produced by immunization with tumor-associated trypsinogen that is isolated from mucinous ovarian cyst fluid. In each assay, one antibody is immobilized onto the walls of polystyrene microtiter strip wells and the other is labeled with an europium(III) chelate. The cross-reaction of each trypsinogen isoenzyme in the assay for the other isoenzyme is less than 1%. The detection limits are 0.1 micrograms/L for trypsinogen-1 and 0.3 micrograms/L for trypsinogen-2. In sera of healthy subjects and patients with extrapancreatic disease the concentration of trypsinogen-1 is higher (median, 21 micrograms/L) than that of trypsinogen-2 (median, 17 micrograms/L), but in acute pancreatitis the ratio is reversed. In acute pancreatitis the concentration of trypsinogen-2 is 50-fold higher than in controls, whereas the difference in trypsinogen-1 concentrations is only 15-fold. The corresponding difference in immunoreactive trypsin measured by a commercially available radioimmunoassay was also only 10-fold.
我们已经针对两种胰蛋白酶原同工酶,即胰蛋白酶原-1和胰蛋白酶原-2(它们也分别被称为阳离子胰蛋白酶原和阴离子胰蛋白酶原)开发了灵敏的时间分辨免疫荧光测定法。这些测定法使用通过用从黏液性卵巢囊肿液中分离出的肿瘤相关胰蛋白酶原免疫产生的单克隆抗体。在每种测定法中,一种抗体固定在聚苯乙烯微量滴定板孔壁上,另一种抗体用铕(III)螯合物标记。每种胰蛋白酶原同工酶在另一种同工酶测定法中的交叉反应小于1%。胰蛋白酶原-1的检测限为0.1微克/升,胰蛋白酶原-2的检测限为0.3微克/升。在健康受试者和胰腺外疾病患者的血清中,胰蛋白酶原-1的浓度(中位数为21微克/升)高于胰蛋白酶原-2的浓度(中位数为17微克/升),但在急性胰腺炎中该比例相反。在急性胰腺炎中,胰蛋白酶原-2的浓度比对照组高50倍,而胰蛋白酶原-1浓度的差异仅为15倍。通过市售放射免疫测定法测得的免疫反应性胰蛋白酶的相应差异也仅为10倍。