Salomo Morten, Gimsing Peter, Nielsen Lars B
Departments of Hematology, Rigshospitalet, University of Copenhagen, Blegdamsvej 9, DK-2100 Copenhagen, Denmark.
Clin Chem. 2002 Dec;48(12):2202-7.
Quantification of free monoclonal light chains in urine [Bence Jones proteins (BJPs)] is used to diagnose multiple myeloma and to evaluate response to treatment. We have developed and evaluated an optimized approach for quantification of BJPs.
High-resolution gel electrophoresis of unconcentrated urine and albumin calibrators was carried out on Sebia's Hydrasys instrument with Hydragel HR agarose gels. After staining with acid violet, the gels were scanned densitometrically. The staining intensities of BJP bands relative to the staining intensities of albumin solutions were used to determine the BJP concentrations. Results for patient samples were compared with conventional agarose gel electrophoresis on concentrated samples.
The relationships between staining intensity and the protein concentrations of albumin and BJPs were linear up to protein concentrations of approximately 2000 mg/L. The detection limit was approximately 20 mg/L. The interassay imprecision (CV) was approximately 8% (n = 23, duplicate analysis), and the results (y) showed a close positive relationship to the comparison method: slope = 0.82 (confidence interval, 0.75-0.88); y-intercept = 34 (-14 to 81) mg/L; n = 29; r(2) = 0.96.
Agarose gel electrophoresis of unconcentrated urine samples together with a series of albumin calibrators followed by acid violet staining and densitometric scanning is sufficiently reproducible and sensitive to quantify clinically relevant BJPs.
尿液中游离单克隆轻链[本周氏蛋白(BJPs)]的定量分析用于诊断多发性骨髓瘤并评估治疗反应。我们开发并评估了一种优化的BJPs定量方法。
使用Sebia公司的Hydrasys仪器及Hydragel HR琼脂糖凝胶对未浓缩尿液和白蛋白校准品进行高分辨率凝胶电泳。用酸性品红染色后,对凝胶进行光密度扫描。通过BJP条带的染色强度与白蛋白溶液染色强度的比值来确定BJP浓度。将患者样本的检测结果与对浓缩样本进行的传统琼脂糖凝胶电泳结果进行比较。
在蛋白质浓度约为2000 mg/L之前,白蛋白和BJPs的染色强度与蛋白质浓度之间呈线性关系。检测限约为20 mg/L。批间精密度(CV)约为8%(n = 23,重复分析),结果(y)与比较方法呈密切正相关:斜率 = 0.82(置信区间,0.75 - 0.88);y轴截距 = 34(-14至81)mg/L;n = 29;r² = 0.96。
对未浓缩尿液样本进行琼脂糖凝胶电泳,结合一系列白蛋白校准品,随后进行酸性品红染色和光密度扫描,对于临床相关BJPs的定量分析具有足够的可重复性和敏感性。