Kuschel Meike, Neumann Tanja, Barthmaier Peter, Kratzmeier Martin
Agilent Technologies Deutschland GmbH, Waldbronn, Germany. :
J Biomol Tech. 2002 Sep;13(3):172-8.
The performance of the Agilent 2100 bioanalyzer, the first commercial lab-on-a-chip system, and the Protein 200 Plus LabChip kit is compared with conventional protein analysis techniques such as SDS-PAGE, Lowry, or Bradford. Lab-on-a-chip technology for protein analysis allows for the integration of electrophoretic separation, staining, destaining, and fluorescence detection into a single process, and for it to be combined with data analysis. The chip-based protein assay allows purity analysis, sizing, and relative quantitation based on internal standards or absolute quantitation based on user-defined standards. The chip-based protein analysis is comparable in sensitivity, sizing accuracy, and reproducibility to SDS-PAGE stained with standard Coomassie. Resolution and linear dynamic range are improved. Absolute quantitation accuracy and reproducibility is improved in comparison to SDS-PAGE and is comparable to batch-based quantitation methods such as Lowry and Bradford. The lab-on-a-chip system has several additional advantages over conventional SDS-PAGE including fast analysis times, reduced manual labor, automated data analysis, and good reproducibility. With such a system, the protein of interest can be tracked during the whole purification procedure, for example, from cell lysates through column fractions to purified proteins.
将安捷伦2100生物分析仪（首个商业化的芯片实验室系统）及Protein 200 Plus LabChip试剂盒的性能与传统蛋白质分析技术（如SDS-PAGE、洛瑞法或考马斯亮蓝法）进行了比较。用于蛋白质分析的芯片实验室技术可将电泳分离、染色、脱色和荧光检测整合到单个过程中，并与数据分析相结合。基于芯片的蛋白质检测可基于内标进行纯度分析、大小测定和相对定量，或基于用户定义的标准进行绝对定量。基于芯片的蛋白质分析在灵敏度、大小测定准确性和重现性方面与用标准考马斯亮蓝染色的SDS-PAGE相当。分辨率和线性动态范围得到了改善。与SDS-PAGE相比，绝对定量准确性和重现性得到了提高，与洛瑞法和考马斯亮蓝法等基于批次的定量方法相当。与传统的SDS-PAGE相比，芯片实验室系统还有几个额外的优点，包括分析时间短、人工操作减少、自动化数据分析和良好的重现性。使用这样的系统，可以在整个纯化过程中跟踪目标蛋白质，例如，从细胞裂解物到柱级分再到纯化的蛋白质。
