Okajima K, Kurobe N, Shimizu K, Kato K
Department of Biochemistry, Institute for Developmental Research, Aichi, Japan.
Clin Chim Acta. 1990 Mar 15;187(3):265-72. doi: 10.1016/0009-8981(90)90111-5.
A sensitive sandwich-type enzyme immunoassay for the aldolase isozyme, A4, was developed using purified antibodies specific to the A subunit of aldolase. The antibodies were raised in sheep being immunized with purified aldolase A4 and then purified by immunoaffinity chromatography on a column of aldolase A4-coupled Sepharose. The assay system consisted of polystyrene balls with immobilized antibody F(ab')2 fragments and the same antibody F(ab')2 fragments labeled with beta-D-galactosidase from Escherichia coli. The assay was sensitive enough to detect 10 pg/tube of aldolase A4. The assay was specific to the A subunit of aldolase (aldolase A). It cross-reacted about 40% to aldolase A3C, 7% to A2C2 and 0.3% to AC3, but not cross-reacted with C4 nor B4. Coefficients of variation in intra- and inter-assay were less than 16%. Serum aldolase A levels were determined in healthy adults, which were about 200 ng/ml. The distribution and concentrations of immunoreactive aldolase A in various human tissues were also determined. High concentrations of aldolase A were found in skeletal muscle, heart muscle, cerebrum and lymphatic tissue.
利用针对醛缩酶 A 亚基的纯化抗体，开发了一种用于醛缩酶同工酶 A4 的灵敏夹心型酶免疫测定法。这些抗体是通过用纯化的醛缩酶 A4 免疫绵羊产生的，然后通过在醛缩酶 A4 偶联的琼脂糖柱上进行免疫亲和层析进行纯化。测定系统由固定有抗体 F(ab')2 片段的聚苯乙烯球和用来自大肠杆菌的β-D-半乳糖苷酶标记的相同抗体 F(ab')2 片段组成。该测定法灵敏度足以检测到 10 pg/管的醛缩酶 A4。该测定法对醛缩酶的 A 亚基（醛缩酶 A）具有特异性。它与醛缩酶 A3C 的交叉反应率约为 40%，与 A2C2 的交叉反应率为 7%，与 AC3 的交叉反应率为 0.3%，但与 C4 和 B4 均无交叉反应。批内和批间变异系数均小于 16%。测定了健康成年人的血清醛缩酶 A 水平，约为 200 ng/ml。还测定了各种人体组织中免疫反应性醛缩酶 A 的分布和浓度。在骨骼肌、心肌、大脑和淋巴组织中发现了高浓度的醛缩酶 A。
