This is a microfluidic immunoblotting device for rapid, parallel profiling of protein components in any biological sample. It enables simultaneous detection of multiple proteins in a single lane of a conventional Western blot membrane with significantly reduced antibody requirements. Since its development over 30 years ago, the Western blot has been an integral analytical technique for the majority of life science research laboratories and it is used as a confirmatory diagnostic for HIV infection. It is a powerful technique that can detect specific antigens in complex biological samples, but it is also time-consuming, labor-intensive, costly in terms of reagents and samples, low throughput, and not easily amenable to multiplexing. A microfluidic device has been designed to In order to improve the versatility and speed of this important biochemical and diagnostic tool.
Microfluidic Device for Immunoblot Development
Researchers at the University of Michigan Medical School designed and validated a microfluidic device that can profile proteins in biological samples with lower resource requirements and higher throughput than traditional methods, but with the same accuracy and sensitivity. After proteins are separated by size using gel electrophoresis and transferred onto a PVDF membrane, the microfluidic device is placed on a dry membrane and secured onto a glass support. This device contains channels that overlay the lanes of a standard immunoblot, and antibody solutions are injected into each channel, such that each lane can be probed with up to five different antibodies at the same time. Using this microfluidic device, the researchers simultaneously profiled the expression and phosphorylation of proteins in the MAPK signaling pathway following LPS stimulation using only two sample lanes. This device minimized the time, resources, and assay variability compared to traditional immunoblotting. There were no issues with leakage or cross-contamination of antibody solutions, and background levels were comparable to traditional methodologies.
- Life science research
- Protein chemistry tool
- Molecular diagnostics and biomarker discovery
- Interfaces with existing technology
- Low cost and high reproducibility
- Reduction in sample and reagent requirements
- Increase in throughput and reliability
- Compatible with several protect detection methodologies