Microfluidic Technology Applied to Protein Sizing and Quantitation
P.Barthmaier, M. Kuschel, T. Neuman
Agilent Technologies, US
Keywords: microfluidics, Protein
Although protein analysis technologies are developing fast, the current standard method is still denaturing gel electrophoresis (SDS-PAGE). Many researchers rely on this traditional method that has not substantially changed in the past 30 years. However, with the increasing focus on proteins, there is a strong demand for automation and higher throughput.
The development of microfluidics offers an alternative for protein analysis and therefore has stimulated a lot of academic and industrial research. Microfluidics allows to actively control fluids in microfabricated channels with a few micrometer in dimension without any moving parts. It aims to integrate several experimental steps into one process. Recently, the first commercial microfluidic system, the Agilent 2100 bioanalyzer, was introduced. The bioanalyzer allows rapid and automated separation of proteins, integrating multiple experimental procedures, such as sample handling, separation, staining, destaining, detection and data analysis.
The performance of this system was compared to conventional protein analysis techniques such as SDS-PAGE, and colorimetric protein quantitation methods (Lowry or Bradford). 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 Coomassie. Resolution and linear dynamic range are improved. Absolute quantitation accuracy and reproducibility is superior to SDS-PAGE and is comparable to Lowry or Bradford. The microfluidic system has additional advantages over SDS-PAGE including fast analysis times, ease of use, automated data analysis, and good reproducibility.
NSTI Nanotech 2003 Conference Technical Program Abstract