Evaluation of a new capillary zone electrophoresis system for the identification and typing of Bence Jones Protein

Michele Mussap, Silvia Ponchia, Martina Zaninotto, Mariacristina Varagnolo, Mario Plebani

Research output: Contribution to journalArticle

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™ 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 κ BJP and 0.0007 g/L for λ 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™ 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.

Original languageEnglish
Pages (from-to)152-159
Number of pages8
JournalClinical Biochemistry
Volume39
Issue number2
DOIs
Publication statusPublished - Feb 2006

Fingerprint

Bence Jones Protein
Capillary Electrophoresis
Electrophoresis
Urine
Proteins
Sepharose
Salts
Gels
Capillary electrophoresis
Agar Gel Electrophoresis
Validation Studies
Dilution
Limit of Detection
Screening
Scanning

Keywords

  • Bence Jones Protein
  • Capillary zone electrophoresis
  • Immunofixation
  • Urine protein electrophoresis

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Evaluation of a new capillary zone electrophoresis system for the identification and typing of Bence Jones Protein. / Mussap, Michele; Ponchia, Silvia; Zaninotto, Martina; Varagnolo, Mariacristina; Plebani, Mario.

In: Clinical Biochemistry, Vol. 39, No. 2, 02.2006, p. 152-159.

Research output: Contribution to journalArticle

Mussap, Michele ; Ponchia, Silvia ; Zaninotto, Martina ; Varagnolo, Mariacristina ; Plebani, Mario. / Evaluation of a new capillary zone electrophoresis system for the identification and typing of Bence Jones Protein. In: Clinical Biochemistry. 2006 ; Vol. 39, No. 2. pp. 152-159.
@article{88790320db1446c2b600dc700631f203,
title = "Evaluation of a new capillary zone electrophoresis system for the identification and typing of Bence Jones Protein",
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™ 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 κ BJP and 0.0007 g/L for λ 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™ 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.",
keywords = "Bence Jones Protein, Capillary zone electrophoresis, Immunofixation, Urine protein electrophoresis",
author = "Michele Mussap and Silvia Ponchia and Martina Zaninotto and Mariacristina Varagnolo and Mario Plebani",
year = "2006",
month = "2",
doi = "10.1016/j.clinbiochem.2005.10.016",
language = "English",
volume = "39",
pages = "152--159",
journal = "Clinical Biochemistry",
issn = "0009-9120",
publisher = "Elsevier Inc.",
number = "2",

}

TY - JOUR

T1 - Evaluation of a new capillary zone electrophoresis system for the identification and typing of Bence Jones Protein

AU - Mussap, Michele

AU - Ponchia, Silvia

AU - Zaninotto, Martina

AU - Varagnolo, Mariacristina

AU - Plebani, Mario

PY - 2006/2

Y1 - 2006/2

N2 - 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™ 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 κ BJP and 0.0007 g/L for λ 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™ 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.

AB - 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™ 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 κ BJP and 0.0007 g/L for λ 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™ 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.

KW - Bence Jones Protein

KW - Capillary zone electrophoresis

KW - Immunofixation

KW - Urine protein electrophoresis

UR - http://www.scopus.com/inward/record.url?scp=32844458992&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=32844458992&partnerID=8YFLogxK

U2 - 10.1016/j.clinbiochem.2005.10.016

DO - 10.1016/j.clinbiochem.2005.10.016

M3 - Article

C2 - 16337932

AN - SCOPUS:32844458992

VL - 39

SP - 152

EP - 159

JO - Clinical Biochemistry

JF - Clinical Biochemistry

SN - 0009-9120

IS - 2

ER -