用于检测人红细胞中胆色素原脱氨酶的酶联免疫吸附测定法

ELISA for measuring porphobilinogen deaminase in human erythrocytes.

作者信息

Lannfelt L, Wetterberg L, Lilius L, Thunell S, Gellerfors P

机构信息

Karolinska Institutet, Department of Psychiatry, St. Goran's Hospital, Stockholm, Sweden.

出版信息

Clin Chim Acta. 1989 Aug 15;183(2):227-37. doi: 10.1016/0009-8981(89)90338-0.

DOI:10.1016/0009-8981(89)90338-0

PMID:2791307

Abstract

An ELISA method has been developed to quantitate human porphobilinogen deaminase in erythrocyte lysate. The antiserum used in the assay was raised against the erythropoietic form of human porphobilinogen deaminase. The IgG fraction was characterized by use of immunoblotting technique, rocket immunoelectrophoresis and immunotitration and shown to be monospecific. The measuring range of the method was from 4 ng to 50 pg. Intra- and inter-assay coefficients of variation were 6% and 7%, respectively. Erythrocyte lysates from 97 apparently healthy individuals were assayed giving a mean erythrocyte porphobilinogen deaminase protein concentration of 150 +/- 28 SD (micrograms/g Hb) and a specific enzyme activity of 750 +/- 140 SD (nkat/g). Eight patients with acute intermittent porphyria were also investigated. A decreased concentration of enzyme protein, i.e. 84 +/- 13 SD (micrograms/g Hb) with a normal specific activity, was found.

摘要

已开发出一种酶联免疫吸附测定（ELISA）方法来定量测定红细胞裂解物中的人胆色素原脱氨酶。该测定中使用的抗血清是针对人胆色素原脱氨酶的造血形式产生的。通过免疫印迹技术、火箭免疫电泳和免疫滴定对IgG组分进行了表征，结果表明其具有单特异性。该方法的测量范围为4纳克至50皮克。测定内和测定间的变异系数分别为6%和7%。对97名明显健康个体的红细胞裂解物进行了测定，结果显示红细胞胆色素原脱氨酶蛋白的平均浓度为150±28标准差（微克/克血红蛋白），比酶活性为750±140标准差（纳摩尔/克）。还对8例急性间歇性卟啉病患者进行了研究。发现酶蛋白浓度降低，即84±13标准差（微克/克血红蛋白），而比活性正常。