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  • Title: An immunoblotting procedure following agarose gel electrophoresis for detection of Bence Jones proteinuria compared with immunofixation and quantitative light chain determination.
    Author: Withold W, Reinauer H.
    Journal: Eur J Clin Chem Clin Biochem; 1995 Mar; 33(3):135-8. PubMed ID: 7605825.
    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.
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