title: An immunoblotting procedure following agarose gel electrophoresis for detection of Bence Jones proteinuria compared with immunofixation and quantitative light chain determination.
abstract: An immunoblotting procedure for the sensitive detection of Bence Jones proteinuria following agarose gel electrophoresis was developed. After immunonephelometric determination of urinary kappa and lambda light chains [employing antisera to human kappa and lambda light chains (free + bound)], urine samples (diluted to 2.5 mg/l kappa and lambda light chains, respectively) were electrophoretically separated using the Paragon system and blotted by capillary diffusion onto nitrocellulose. Rabbit anti-human kappa and lambda light chains reacted to kappa and lambda light chains attached to the membrane. Goat anti-rabbit IgG alkaline phosphatase conjugate was employed as detection system. The detection limit of the immunoblotting procedure (monoclonal component, as determined by serial dilutions) was 0.3 mg/l urine. Among 65 urine specimens received for routine testing for Bence Jones proteinuria, 32 monoclonal components (in 20 urine samples) were found by immunoblotting compared with 10 monoclonal components (in 9 urine samples) detected by immunofixation. In only 5 out of these 65 urine samples a kappa/lambda ratio (as determined immunonephelometrically) < 1 or > 5.2 (decision limits for discriminating between monoclonal and polyclonal urinary light chains; Boege F, Koehler B, Liebermann F. Eur J Clin Chem Clin Biochem 1990; 28:37-42) was observed. In conclusion, the immunoblotting method is superior to both immunofixation and immunonephelometry with respect to the diagnostic sensitivity for detection of Bence Jones proteinuria.
authors.affiliation: Institute für Klinische Chemie und Laboratoriumsdiagnostik, Heinrich-Heine-Universität Düsseldorf, Germany.
authors.forename: W
authors.initials: W
authors.lastname: Withold
authors.forename: H
authors.initials: H
authors.lastname: Reinauer
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D001536:Bence Jones Protein / Q000652:urine*; D004587:Electrophoresis, Agar Gel / Q000379:methods; D006801:Humans; D015151:Immunoblotting / Q000379:methods*; D007145:Immunoglobulin kappa-Chains / Q000652:urine; D007146:Immunoglobulin lambda-Chains / Q000652:urine; D009391:Nephelometry and Turbidimetry / Q000379:methods; D011233:Precipitin Tests / Q000379:methods
chemical_list: D000911:Antibodies, Monoclonal; D007145:Immunoglobulin kappa-Chains; D007146:Immunoglobulin lambda-Chains; D001536:Bence Jones Protein