Background This report describes a nephelometric assay for measuring complement 3d (C3d).Methods C3d was separated from C3 and the other C3 split products by polyethylene glycol precipitation. The supernatant containing C3d was then measured by rate nephelometry in a Beckman Immage rate nephelometer using a specific antibody. Calibration was performed with dilutions of a standard, which was obtained by incubating a serum pool at 37degreesC for 7 days. This incubation step allowed the activation and breakdown of C3 into C3d.Results The assay was linear across the normal and pathological range. Precision studies showed within-run and between-run coefficients of variation of <5% and <6%, respectively. The nephelometric results are comparable with those obtained by radial immunodiffusion. Neither haemoglobin nor lipids interfere with the assay.

We report age-related reference intervals for capillary zone electrophoresis for children between 1 and 14 years of age.

It has been shown that benzethonium chloride produces linear mixed-type inhibition of choline esterase and acetylcholine esterase. These enzymes also show-reagent-carry-over inhibition if the enzyme activities are measured in plastic cuvettes in which previously protein has been determined by the alkaline benzethonium chloride method. Choline esterase is about 10-fold more sensitive to benzethonium chloride than acetylcholine esterase. With acetylthiocholine as substrate Michaelis-Menten constants for choline esterase and acetylcholine esterase are 85 mu mol/l and 102 mu mol/l, respectively. Carry-over inhibitory effect of benzethonium chloride can be avoided by washing the cuvettes, after protein determination by the benzethonium chloride method, with 5 ml/l Triton X-100, 5 ml/l Tween 20 or 10 g/l sodium dodecyl sulphate. The latter has a disadvantage in that it precipitates out at low temperatures. The dry slide method (Johnson & Johnson) for serum choline esterase is free of the inhibitory effect until the concentration of benzethonium chloride in the sample reaches about 200 mu mol/l.

Phosphate concentrations were determined in 52 cases of paraproteinemia. The unmodified acidic ammonium molybdate method produced 19% spuriously high results. The false increase of phosphate concentration was attributable to formation of precipitate in the reaction mixture. The precipitate was formed by interaction between immunoglobulins and the unmodified acidic ammonium molybdate reagent. The magnitude of interference bore no relation to the type, concentrations, or isoelectric point of the paraproteins or to the presence or absence of free light chains. Diluting the sample to approximately 40 g/L total protein reduced but did not always eliminate the interference. In some cases paraprotein concentration as low as 8 g/L falsely increased plasma phosphate results. Apparently, only IgG and IgM but not IgA paraproteins produced the interference. Deproteination by ultrafiltration or by treatment with trichloroacetic acid removed the interference. The Kodak slide method and the new modified Boehringer Mannheim phosphate test were found to be interference-free. However, in some cases the latter new formulation is sensitive to substantial changes in ionic concentration of the reaction mixture.