Electrophoresis underestimates the concentration of polyclonal immunoglobulins in serum

The concentration of gamma-migrating immunoglobulins in blood serum was measured in 200 healthy blood donors by two methods, nephelometry and electrophoresis. Nephelometric values were calculated as IgG + IgM + 1/2 IgA because immunoglobulin G (IgG) and immunoglobulin M (IgM) migrate entirely within the gamma region and immunoglobulin A (IgA) is split between the gamma and beta regions. Electrophoretic values were calculated as % gamma x total protein. Gels were scanned with two different densitometers, providing two sets of electrophoretic data. In each case, the correlation between nephelometry and electrophoresis was very good (r = 0.94 and r = 0.96). However, electrophoresis consistently gave lower results than nephelometry (mean = 2.3 g/L and 4.8 g/L lower, respectively). The disparity between these methods was greater as the concentration of immunoglobulins increased. It is concluded that (1) electrophoresis gives a relative but not absolute value of immunoglobulin concentration in serum; (2) nephelometric and electrophoretic values cannot be used interchangeably; and (3) reference ranges for electrophoresis must take into account the densitometer used.