Effect of ampicillin-sulbactam on clinical capillary zone electrophoresis of serum proteins

BACKGROUND: Capillary zone electrophoresis (CZE) is a well-accepted automated method used to separate serum proteins and detect monoclonal components. CZE uses ultraviolet detection at 214 nm to directly quantify proteins via peptide bonds. Any substance that absorbs at 214 nm and is present in serum can potentially interfere with CZE analysis. This has been reported for radio-contrast media and antibiotics. METHODS: Here we describe a peak on the anode side of the alpha(2)-globulin fraction caused by the antibiotic ampicillin-sulbactam (Unacid). RESULTS AND CONCLUSIONS: Extra peaks that can be misinterpreted as monoclonal components can be present in almost all electrophoretic fractions of CZE. Immunosubtraction or immunofixation is always required to rule out these conditions.     

Separation of serum proteins by automated capillary zone electrophoresis.

Capillary zone electrophoresis (CZE) of serum proteins is increasingly gaining impact in clinical laboratories. Two automated multichannel instruments dedicated to the separation of serum proteins have become available over the last 6 years, the Paragon CZE 2000 (Beckman Coulter, CA, USA) and, more recently, the Capillarys (Sebia, France). This review focuses on the performance of these commercial instruments to separate serum proteins in a clinical laboratory setting. The utility of CZE to recognize various dysproteinemias and to detect and identify monoclonal proteins will be described and systematically reviewed. The reader will be provided with a summation and an understanding of CZE-specific interference.     

全自动多通道毛细管区带电泳法在血清蛋白分析中的临床应用

目的用毛细管区带电泳(CE)技术进行血清蛋白检测结果与琼脂糖凝胶电泳(AGE)技术的检测结果进行比较分析以探讨CE的临床应用。方法使用全自动毛细管电泳仪(法国Sebia公司的4·51版本Capillarys电泳仪)及相应试剂,在7kV电压、35·5℃、pH10的缓冲液条件下,在15·5cm×25·0μm硅毛细管内进行血清蛋白电泳,用200nm波长检测各种蛋白质的百分比浓度。结果与AGE的结果比较相关性,进行CE的精密度、线性检测及干扰实验。CE和AGE比较,白蛋白、α1-球蛋白、α2-球蛋白、β-球蛋白及γ-球蛋白的相关系数分别为0·929、0·924、0·841、0·789和0·926,P<0·001相关性好,无统计学意义。蛋白质含量3个水平不同标本的批内、批间结果,白蛋白、α1-球蛋白、α2-球蛋白、β1-球蛋白、β2-球蛋白和γ-球蛋白的变异系数(CV)值均小于10·08%精密度好。血清总蛋白浓度在71·10~11·16g/L范围内各种蛋白线性关系良好,白蛋白最低检出量为6·87g/L,γ-球蛋白为1·78g/L。血清总胆红素含量≤144·7mmol/L、甘油三酯含量≤8·18mmol/L时对CE结果无干扰,纤维蛋白原和血红蛋白对CE结果有干扰。结论CE技术是一种灵敏度高、精确度好的血清蛋白检测技术,随着设备自动化的实现,分析操作将更简便、快速,适于临床化学实验室使用。     

Reference values for the five electrophoretic serum protein fractions in Caucasian children by capillary zone electrophoresis.

We report age-related reference intervals for capillary zone electrophoresis for children between 1 and 14 years of age.     

Linearity and detection limit in the measurement of serum M-protein with the capillary zone electrophoresis system Capillarys.

We studied the linearity and detection limits of the capillary zone electrophoresis system Capillarys in the measurement of serum monoclonal protein. Three monoclonal proteins with different isotypes and electrophoretic migrations were diluted with a hypo-gamma-globulinemic polyclonal serum pool. Mathematical linearity was observed for all monoclonal protein isotypes in the ranges studied without removing the polyclonal gamma-globulin background. Theoretical concentrations of 0.43, 0.89 and 0.33 g/L for monoclonal proteins immunoglobulin (Ig)G, IgA and IgM, respectively, gave a discernible spike by Capillarys, although they were measured as 0.76, 1.09 and 0.76g/L, respectively. We observed overestimation of monoclonal protein inversely correlated to concentrations below 15 g/L. All these limitations have to be taken into account when monitoring monoclonal proteins, because the loss of linearity and the protein background may hide an increase in concentration at low levels.     

血清蛋白质组学研究中乙腈沉淀法去除清蛋白的实验研究

目的 建立去除人血清清蛋白的乙腈沉淀法并研究其去除效果.方法 血清经除脂后按血清:水:乙腈=1:2:4.5(v/v/v)比例沉淀分离高丰度清蛋白,用SDS-PAGE验证去除效果和重复性.分别对4名体检健康者和非小细胞肺癌患者去除清蛋白前后的血清凝胶电泳图像进行分析.结果 经本法去除后血清清蛋白平均浓度由(50.65 4...     

Ultraviolet-absorbing organic anions in uremic serum separated by capillary zone electrophoresis, and quantification of hippuric acid

Organic anions accumulated in blood serum of patients with chronic renal failure were separated by a novel technique: closed-system capillary zone electrophoresis (CZE) in a pH6 carrier-electrolyte system. Hippuric acid (HA), p-hydroxyhippuric acid, and uric acid were identified by their co-elution with standards prepared in ultrafiltered normal serum and by comparison with the corresponding ultraviolet-detected peaks positively identified in the HPLC analyses. Analysis time for the entire profile is 8 min. Repeatabilities (CVs) of CZE migration times and peak areas of the three acids in serum samples were about 0.7% and 6%, respectively. We quantified HA in 10 ultrafiltered uremic serum samples and compared results with those by a previously described HPLC procedure. The very good agreement further supports the identification of hippuric acid. Accuracy and precision of the CZE method were similar to those for the HPLC gradient-elution method, but analysis time for HA (8 min) is much less than by HPLC (90 min). Our technique is very suitable for selective, rapid analysis for (ultraviolet-absorbing) anionic constituents in ultrafiltered uremic serum, without any sample pretreatment.     

高效毛细管电泳法在血清氨甲蝶呤浓度检测中的应用

目的建立快速检测人血清中氨甲蝶呤药物浓度的方法。方法用高效毛细管电泳技术，选用直径75μm，总长60cm，有效长度50．5cm熔融石英毛细管，以75mmoL／L，pH7．4的磷酸盐为电泳缓冲液，检测波长为306nm，在25kV电压下的快速分离检测氨甲蝶呤，并对此方法进行方法学评价。结果氨甲蝶呤的出峰时间在10min内，线性范围在1.1—1100．0μmol／L间，最低检出浓度为0．55μmoL／L，回收率为88．2％-98．2％，批间精密度为5．4％，批内精密度为4．2％。结论高效毛细管电泳方法具有快速、准确且成本低的特点，为临床和科研中测定血清氨甲蝶呤提供了一种有效的手段。