Response-surface-optimized, zinc-enhanced assay for serum alkaline phosphatase.
An optimized assay for alkaline phosphatase (EC 3.1.3.1) is reported. A discrete analyzer, the DuPont Automatic Clinical Analyzer (aca), was used for this study. The assay is based on results of response-surface experimental co-optimization techniques, and response is enhanced over the present aca assay. A key feature is the incorporation of zinc ions, both to fully optimize the assay and to reduce the sensitivity of measured activity to zinc-binding impurities in the buffer, 2-amino-2-methyl-1-propanol. In addition, a simple technique is described for measuring relative concentrations of zinc-binding impurities in this buffer. These features should be considered in the design of any assay for alkaline phosphatase that is based on p-nitrophenyl phosphate as substrate and 2-amino-2-methyl-1-propanol as buffer.
D000469:Alkaline Phosphatase / Q000097:blood*; D001322:Autoanalysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008274:Magnesium / Q000494:pharmacology; D013053:Spectrophotometry / Q000379:methods; D015032:Zinc / Q000494:pharmacology*
D000469:Alkaline Phosphatase; D008274:Magnesium; D015032:Zinc