ELISA for measuring porphobilinogen deaminase in human erythrocytes.
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.
Lannfelt
L
L
Karolinska Institutet, Department of Psychiatry, St. Goran's Hospital, Stockholm, Sweden.
Wetterberg
L
L
Lilius
L
L
Thunell
S
S
Gellerfors
P
P
D000642:Ammonia-Lyases / Q000097:blood*; D000818:Animals; D004797:Enzyme-Linked Immunosorbent Assay; D004912:Erythrocytes / Q000201:enzymology*; D006801:Humans; D011163:Hydroxymethylbilane Synthase / Q000097:blood* / Q000276:immunology; D007106:Immune Sera / Q000276:immunology; D015151:Immunoblotting; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000276:immunology; D011817:Rabbits; D012015:Reference Standards; D015203:Reproducibility of Results
D007106:Immune Sera; D007074:Immunoglobulin G; D011163:Hydroxymethylbilane Synthase; D000642:Ammonia-Lyases