Objectives: Capillary electrophoresis has recently emerged as a new sensitive technique for the separation of urinary proteins. We evaluated a new method for Bence Jones Protein (BJP) detection and characterization on native urine samples by the Paragon CZE (TM) 2000 system. To avoid interference in electrophoretic separation, urine samples were preliminarily treated for the selective removal of interfering salt particles. Design and methods: The evaluation was done on a total of 350 fresh 24-h urine samples. The salt particle removal consisted of a manual chromatographic separation, optimized in the course of our evaluation. Capillary zone urinary protein electrophoresis (CZ-UPE) was compared with conventional high-resolution electrophoresis on an agarose gel, while capillary immunosubtraction (CZU-IFE) was compared with agarose gel immunofixation. Results: After finding a consistent protein loss in eluates, the preanalytical treatment was optimized by changing sample dilution and eluate collection. The within- and between-run imprecision values for monoclonal peaks corresponding to BJP ranged from 0.4-12.2% to 3.3-6.3%, respectively. The detection limit for BJP, defined as the lowest measurable monoclonal peak on CZ-UPE, was 0.0012 g/L for kappa BJP and 0.0007 g/L for lambda BJP. CZ-UPE and CZU-IFE sensitivities were significantly lower in urine samples with a total protein level <= 100 mg/L (67% and 78%, respectively) compared to those with total protein > 100 mg/L (92% and 94%, respectively). Comparison between BJP measurements obtained from densitometric scanning with those from absorbance tracing showed a correlation coefficient of 0.994 and a bias of 29.8 mg/L. Conclusions: Paragon CZE (TM) 2000 can be introduced in routine for screening and typing of BJP; in urine samples with a total protein level > 100 mg/L, the performance is consistent with results from published validation studies on CZE applied to serum samples. (c) 2005 The Canadian Society of Clinical Chemists. All rights reserved.
Evaluation of a new capillary zone electrophoresis system for the identification and typing of Bence Jones Protein
MUSSAP, MICHELE;ZANINOTTO, MARTINA;PLEBANI, MARIO
2006
Abstract
Objectives: Capillary electrophoresis has recently emerged as a new sensitive technique for the separation of urinary proteins. We evaluated a new method for Bence Jones Protein (BJP) detection and characterization on native urine samples by the Paragon CZE (TM) 2000 system. To avoid interference in electrophoretic separation, urine samples were preliminarily treated for the selective removal of interfering salt particles. Design and methods: The evaluation was done on a total of 350 fresh 24-h urine samples. The salt particle removal consisted of a manual chromatographic separation, optimized in the course of our evaluation. Capillary zone urinary protein electrophoresis (CZ-UPE) was compared with conventional high-resolution electrophoresis on an agarose gel, while capillary immunosubtraction (CZU-IFE) was compared with agarose gel immunofixation. Results: After finding a consistent protein loss in eluates, the preanalytical treatment was optimized by changing sample dilution and eluate collection. The within- and between-run imprecision values for monoclonal peaks corresponding to BJP ranged from 0.4-12.2% to 3.3-6.3%, respectively. The detection limit for BJP, defined as the lowest measurable monoclonal peak on CZ-UPE, was 0.0012 g/L for kappa BJP and 0.0007 g/L for lambda BJP. CZ-UPE and CZU-IFE sensitivities were significantly lower in urine samples with a total protein level <= 100 mg/L (67% and 78%, respectively) compared to those with total protein > 100 mg/L (92% and 94%, respectively). Comparison between BJP measurements obtained from densitometric scanning with those from absorbance tracing showed a correlation coefficient of 0.994 and a bias of 29.8 mg/L. Conclusions: Paragon CZE (TM) 2000 can be introduced in routine for screening and typing of BJP; in urine samples with a total protein level > 100 mg/L, the performance is consistent with results from published validation studies on CZE applied to serum samples. (c) 2005 The Canadian Society of Clinical Chemists. All rights reserved.
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