title: Sensitive enzyme immunoassay for human aldolase A.
abstract: 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.
authors.affiliation: Department of Biochemistry, Institute for Developmental Research, Aichi, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Okajima
authors.forename: N
authors.initials: N
authors.lastname: Kurobe
authors.forename: K
authors.initials: K
authors.lastname: Shimizu
authors.forename: K
authors.initials: K
authors.lastname: Kato
mesh_terms: D005634:Fructose-Bisphosphate Aldolase / Q000032:analysis*; D006801:Humans; D007124:Immunoenzyme Techniques; D007527:Isoenzymes / Q000032:analysis*
chemical_list: D007527:Isoenzymes; D005634:Fructose-Bisphosphate Aldolase