Immunochemical determination of human immunoglobulins: use of kinetic turbidimetry and a 36-place centrifugal analyzer

Abstract

We describe the measurement of human immunoglobulins IgG, IgA, and IgM in diluted sera, with diluted commercial monospecific antisera, with use of a 36-place centrifugal analyzer (RotoChem IIa; American Instrument Co., Silver Spring, MD 20920). The assay involves a two-point kinetic turbidimetric technique. The changes in absorbance at 340 nm between 10 and 255 s are taken for standard curve construction with use of a computer-generated cubic least-squares fit. Patients' samples are quickly calculated from the stored curve. Multiple time-interval readings are taken to observe the reaction kinetics. The lowest detectable concentrations are: IgG, 5 mg/L; IgA, 20 mg/L; and IgM, 20 mg/L. Correlation with radial immunodiffusion was excellent. Precision, accuracy, linearity, and sensitivity were very acceptable. Antigen or antibody excess can be easily detected. The accuracy of the proposed method when measuring idiotypic monoclonal proteins is greater than radial immunodiffusion. Up to 30 patients' samples can be analyzed at one time, and calculation of test results by use of the computer program is efficient and rapid.