Development of analytical methods for human fluids by capillary electrophoresis

毛细管电泳人体体液分析方法的开发

• 批准号: 10672179
• 负责人: UJI Yoshinori
• 金额: $ 2.11万
• 依托单位: Kumamoto University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 1999
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10672179/
• 关键词: capillary electrophoresis; LD isoenzymes; Alp isoenzymes; immunofixation by subtraction; serum protein analysis; アイソザイム分析

I have developed and evaluated the analytical method for human fluids by capillary eletrophoresis(CE). In the serum protein fractions, good linear correlation was observed between CE and cellulose acetate membrane electrophoresis (CA). The substantially higher values found for the α-1G fraction with a when compared with CA is related to the fact bothα-1 antitripsin and α-1 acidglycoprotein are quantitated by direct absorption with CE whereas with CA, the high sialic acid content of α-1-acid glycoprotein interferes with the binding of dyes used to quantitated the protein fraction. Reference ranges for albumin, α-1-G, and A/G were significantly different between CE and CA. No fiblinogen peak can be found in plasma sample with CE, because the concentration of fiblinogen is low in plasma samples. No interfering substances were observed both CA and CE. In the identification of serum M-protein by a immunofixation by subtraction (CE-IFEs), concordance studied between /IFE/s and agarose gel im … More munofixation eletrophoresis showed good agreement in identifying 30 monoclonal gammapathy patient samples. However, missed typing was observed containing very small monoclonal component or a second monoclonal hidden under a larger one or very close to one typed. Classical method, like immunoelectrophoresis or immunofixation, should be used for these samples. Separation and quantitative estimation of the isoenzymes of lactate dehydrogenase(LD) and alkaline phosphatase (ALP) isoenzyme in serum were accomplished. A uncoated fused sillica capillary column 50 cm(long) X 75mm( I.D. ) and substrate containing running buffer (L-lactic acid and NAD for LD isoenzymes,α-napthyl phospate and MgClィイD22ィエD2 for ALP isoenzymes) were used. The resulting product NADH for LD andα- napthol for ALP were detected at 340 nm. The results obtained by the proposed method both LD and ALP were correlated well with CA method. In conclusion, proposed analytical method by capillary eletrophoresis system is sensitive, precise and easy for clinical laboratory use. Less

我开发并评价了毛细管电泳法分析人体体液的方法。在血清蛋白质组分中，毛细管电泳法与醋酸纤维素膜电泳法具有良好的线性相关性。与CA相比，α-1G组分的a值高得多，这与α-1抗Tripsin和α-1酸性糖蛋白都是通过CE直接吸收来定量的事实有关，而对于CA，α-1酸性糖蛋白的高唾液酸含量干扰了用于定量蛋白质组分的染料的结合。白蛋白、α-1-G和A/G的参考值范围在CE和CA之间有显著差异。由于血浆中纤维蛋白原的浓度较低，毛细管电泳法在血浆样品中未发现纤维蛋白原峰。CA和CE均未观察到干扰物质。消减免疫固定(CE-IFE)鉴定血清M-蛋白中/IFE/S与琼脂糖凝胶…的一致性研究更多的免疫固定电泳法在鉴定30例单克隆性伽马病患者样本中表现出很好的一致性。然而，观察到遗漏的分型包含非常小的单抗成分或隐藏在较大的或非常接近分型的单抗下面的第二个单抗。对于这些样品，应采用免疫电泳法或免疫固定法等经典方法。完成血清乳酸脱氢酶(LD)和碱性磷酸酶(ALP)同工酶的分离和定量测定。未涂层熔融硅胶毛细管柱50厘米(长)×75毫米(内径)含运行缓冲液的底物(乳酸脱氢酶L-乳酸和乳酸脱氢酶NAD，碱性磷酸酶脱氢酶α-napthyl phopate和碱性磷酸酶脱氢酶ィイD22ィエD2)。在340 nm处检测到LD的产物NADH和碱性磷酸酶的α-napthol。用该方法得到的LD和ALP结果与CA法具有很好的相关性。总之，毛细管电泳法是一种灵敏、准确、易于临床实验室使用的分析方法。较少

项目成果

Uji,Y., et al.: "Evalution of Beckman Paragon CZE 2000 system; SPE and IFE/s"Clin Chem. A19-A19. (1998)

Uji,Y. 等人：“Beckman Paragon CZE 2000 系统的评估；SPE 和 IFE/s”Clin Chem。

Uji,Y.,et al.: "Evaluation of Beckman Paragon CZE2000 system; SPEs and IFE." Clin Chem. 44. A19 (1998)

Uji,Y.,et al.：“Beckman Paragon CZE2000 系统的评估；SPE 和 IFE。”

Uji,Y.: "Application of Capillary electrophoresis in clinical laboratory (serum protein fraction, serum M protein and isoenzymes)"Journal of Kyushu Clinical Chemistry. 9. 44-51 (1999)

Uji,Y.：“毛细管电泳在临床实验室中的应用（血清蛋白组分、血清M蛋白和同工酶）”九州临床化学杂志。

宇治義則: "キャピラリー電気泳動法の臨床検査への応用(血清蛋白分画、血清M蛋白の同定・血液アイソザイムの測定について)"日本臨床化学会九州支部会誌. 9. 44-51 (1999)

Yoshinori Uji：“毛细管电泳在临床检测中的应用（血清蛋白分级、血清M蛋白的鉴定和血液同工酶的测量）”日本临床化学学会九州分会杂志9. 44-51（1999）。