Objectives: Immunoglobulin paraproteins can interfere with multiple chemistry assays. We want to investigate the mechanisms of immunoglobulin interference.
Design and methods: Serum samples containing paraproteins from the index patient and eight additional patients were used to investigate the interference with the creatinine and total protein assays on the Beckman Coulter AU5400/2700 analyzer, and to determine the effects of pH and ionic strength on the precipitation of different immunoglobulins in these patient samples.
Results: The paraprotein interference with the creatinine and total protein assays was caused by the precipitation of IgM paraprotein in the index patient's samples under alkaline assay conditions. At extremely high pH (12-13) and extremely low pH (1-2) and low ionic strength, paraprotein formed large aggregates in samples from the index patient but not from other patients.
Conclusions: The pH and ionic strength are the key factors that contribute to protein aggregation and precipitation which interfere with the creatinine and total protein measurements on AU5400/2700. The different amino acid sequence of each monoclonal paraprotein will determine the pH and ionic strength at which the paraprotein will precipitate.
Keywords: AMR, analytical measurement range; AU2700, Beckman Coulter AU2700 analyzer; AU5400, Beckman Coulter AU5400 analyzer; Assay conditions; CV, coefficient of variation; Cobas 8000, Roche Cobas 8000; Creatinine; HCl, hydrochloric acid; HDL-C, high density lipoprotein cholesterol; IFE, immunofixation electrophoresis; IRB, Institutional Review Board; IgM paraprotein; Interference of chemistry assays; LPL, lymphoplasmacytic lymphoma; Monoclonal protein; NaCl, sodium chloride; NaOH, sodium hydroxide; SPEP, serum protein electrophoresis; WM, Waldenström's macroglobulinemia; pH induced.