Detection of Bence-Jones protein by isoelectric focussing of unconcentrated urine followed by nitrocellulose blotting and immunoperoxidase staining

Abstract

Urine specimens from 164 patients sent to the laboratory for testing for Bence-Jones proteinuria were investigated using a new procedure. The protein in the untreated urine was subjected to isoelectric focussing in an agarose gel, transferred to a nitrocellulose membrane by blotting, and then stained by an immunoperoxidase technique for either immunoglobulin kappa or lambda chains. This technique was compared with a routine procedure for the detection of immunoglobulin light chains involving concentration by ultrafiltration, electrophoresis and then immunofixation. The new technique achieved a much increased rate of detection of Bence-Jones proteinuria. Among 51 patients known to have myeloma or macroglobulinaemia, Bence-Jones proteinuria was detected in 35 cases with the new procedure and in only 27 by the conventional method. In 28 patients with paraproteinaemia without other evidence of myeloma, macroglobulinaemia, leukaemia or lymphoma, 12 instances of Bence-Jones proteinuria were discovered with the new procedure, 10 of which were missed by the conventional method. The improved efficiency of detection is attributed to the high resolution of isoelectric focussing and the avoidance of protein loss from adsorption on to ultrafiltration membranes.