Summary:A new method for phenotyping human serum arylesterase (EC 3.1.1.2) is described and evaluated. The aromatic esters, phenyl acetate and 4-nitrophenyl acetate, were compared as substrates for spectrophotometric measurement of arylesterase activity. A method for arylesterase phenotyping, based upon inhibition of the enzymatic hydrolysis of 4-nitrophenyl acetate by phenyl acetete, was developed. The method was applied to serum samples from 158 blood donors and showed a distinct separation of the three phenotypes defined by a reference method based on the ratio of paraoxonase activity to arylesterase activity using paraoxon and phenyl acetate as substrates. The method was adapted to a Cobas-Fara centrifugal analyser.
The occurrence of raised liver-derived enzymes was frequent in the Danish population sample and associated with moderate self-reported alcohol consumption adjusted for BMI and smoking.
The influence of body weight, height, age and sex on plasma cholinesterase activity (ChE) in 650 males and 437 females with ChE-1 phenotype U (genotype ChEuChEu or ChEuChEs) or UA (genotype ChEuChEa) was studied in a multiple regression model. ChE was not influenced by age (p > 0.01), but, like other liver synthesized plasma enzymes, highly (p < 0.001) influenced by body weight and height. In a logarithmic scale ChE followed a linear model (R = 0.535, p < 0.001) with randomly distributed residuals, InChE = 3.286-0.308 x ChE-1 phenotype-0.104 x sex + 0.00765 x weight - 0.00723 x height (U = 1, UA = 2; male = 1, female = 2; kg; cm). A simplified model based on body-mass index (BMI = weight divided by squared height, kg/m2), InCHE = 2.016-0.308 x ChE-1 phenotype - 0.091 x sex + 0.0230 x BMI, showed the same goodness-of-fit (R = 0.533). In a non-logarithmic scale both multiple regression models failed to fit cases with high ChE activity. A model for a 'standardized' plasma ChE in which the effects of ChE-1 phenotype, sex, body weight and height are eliminated, is proposed to compare ChE in unmatched population groups when using this enzyme activity as a biomarker in environmental or occupational medicine.
Summary: We developed an enzyme immunoassay (ELISA) for quantitation of plasma cholinesterase substance concentrations in native plasma or serum samples. The ELISA assay is based on polyclonal (rabbit) antihuman cholinesterase and a highly specific monoclonal (mouse) antibody, with a commercially available peroxidase-conjungated (rabbit) antibody directed against mouse immunoglobulins as the signal carrier. The detected serum cholinesterase substance concentrations (mean: 4.51 mg/1, SD: 0.90 mg/1) in randomly selected serum samples from 33 healthy individuals were closely and linearly related to the corresponding catalytic activity concentrations.
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