DROPLET-ELECTROPHORESIS DEVICES AND RELATED METHODS

DROPLET-ELECTROPHORESIS DEVICES AND RELATED METHODS

Researchers at UC Berkeley have developed a novel method for comprehensively profiling the proteomes of single cells.

Genomics (DNA) and transcriptomics (RNA) sequencing is one of the cornerstones of our modern-day understanding of the genetic blueprint of life. Furthermore, sequencing and analysis of the genomes of single cells is considered one of the seminal advances in biotechnology in the past quarter-century. Rather than sequencing bulk nucleic acids and getting a quantitative signal of gene expression in an entire population of cells, single cell sequencing allows heterogenous populations of cells to be assayed. This in turn permits the detection of novel and/or rare cell types in a massively multiplexed format. However, methods to assay expression at the protein level are limited and those that do exist have large caveats associated with their use. Immunoassays and mass spectrometry are two most used proteomics methods. However, neither assay provides comprehensive coverage of the proteomes of single cells. Specifically, these assays commonly miss key proteoforms including isoforms, splice variants, and combinations of post translational modifications, all of which are essential for understanding the biology of these proteins in single cells. There is a need in the biomedical community for the development of methods that comprehensively cover all proteoforms in single cells.

Stage of Research

To provide a global snapshot of the entire proteome of single cells, the inventors have created a novel method called DropBlot. This method employs a droplet electrophoresis device that effectively and efficiently combines droplet microfluidics with single cell immunoblotting. In short, cells are deposited into the droplet electrophoresis device and are subsequently encapsulated in water and oil droplets and lysed. From there, protein isoforms electromigrate out of the droplet and undergo protein sizing in a polyacrylamide gel. This then allows for immunoblotting to detect proteins in single cells. This method represents a major step forward in our ability to detect and understand the proteomes inside single cells and has myriad clinical and laboratory applications.

Applications

Investigation of the secretion or expression of proteins or chemokines during communication can help understand the underlying mechanism of human biology or diseases
DropBlot can be used to profile proteins or chemokines during the timed dosing of a drug which can help evaluate cell qualities and estimate the treatment prognosis.

Advantages

Able to operate on fixed clinical samples, which has not been possible with other available methods
DropBlot can simultaneously pair the target cells or particles, monitor the communication

process, and resolve the secreted chemokines or proteins

Stage of Development

Research- in vitro

Publications

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