Enabling Technologies for Biomedical Research

ETBR invents and develops technologies and related platforms that will enable biomedical researchers to detect and identify molecules that previously cannot be analyzed rapidly and economically. In specific, ETBR develops Nano Open Tubular Liquid CHromatography (NOTLC) and related instruments and intellectual properties for high-speed, high-throughput and/or high accuracy proteomic and single cell analysis.

 

Effective analytical tools for single-mammalian-cell analysis (~10 micrometers in diameter) are in high demand. In multicellular organisms, all cells originate from a single zygote, which, through tightly regulated programs of proliferation and differentiation, gives rise to diverse cell types. Dysregulation of these processes in individual "renegade" cells can lead to diseases such as cancer, neurological disorders, and developmental conditions.

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Growing evidence suggests that cellular heterogeneity within seemingly identical populations plays a crucial role in survival and disease progression. Conventional technologies that rely on averaged population responses obscure critical differences between individual cells. To fully understand cell-to-cell variability, it is essential to analyze individual cells from their live state to cell lysate. Such analyses are vital for advancing personalized medicine and therapy development.

However, no existing tools have satisfactorily met this need—until now. Professor Liu's Nano Open Tubular Liquid Chromatography (NOTLC) technique is poised to overcome this challenge, offering a groundbreaking solution for high-precision single-cell analysis.

 

High performance liquid chromatography (HPLC) is the most powerful analytical technique in chemical separation and analysis; it can analyze ~80% of all known compounds and has been widely used for biomedical research, pharmaceutical discovery, clinical diagnosis, environmental analysis, etc. It has currently encountered major challenges (e.g., limited sample, inadequate detection sensitivity and wide dynamic range, etc.) for proteomic and single cell analysis.

 

Since the successful completion of the Human Genome Project that mapped the whole human genome, in which Professor Liu and co-workers developed a DNA sequencer for this project, a massive number of genomic markers have been identified and are being applied to medical sciences. Proteomics has emerged as a rapidly advancing field, particularly in therapeutic research. Unlike genomics, which provides a static blueprint of an organism, proteomics offers deeper insights into biological structure and function by analyzing dynamic protein expression. However, proteomics is far more complex than genomics, as protein expression varies over time and in response to environmental conditions. While the human genome is estimated to encode approximately 26,000 to 31,000 proteins, the total number of proteoforms is believed to range from 10^6 to 10^9, highlighting the immense complexity of the proteome.

 

Chromatography is a powerful tool that can separate proteins from complex mixtures and can analyze large and fragile biomolecules. When combined with mass spectrometry, it can be used for determining the peptides in the mixtures. Chromatography can help researchers discover novel biomarkers and understand the mechanisms of carcinogenesis according to the modifications of proteins.

 

However, this technique faces several serious challenges:

1) Limited sample quantity. Often, the same sample may be used for several analyses, and each analysis may need to be performed multiple times in order to get statistical information. Most proteomic samples do not have adequate quantities for all these analyses. 

2) Inadequate detection sensitivity. Some proteoforms may have less than 1000 copies in a cell; current chromatography cannot analyze it.

3) Wide dynamic range. While some proteoforms may have only 1000 copies, some other proteoforms may have 10^9~10^11 copies. Current chromatography cannot handle this wide dynamic range.

 

All these problems can be alleviated using NOTLC.

 

NOTLC is a format of chemical analysis technique that uses very narrow open tubular columns. The concept of open tubular columns was first introduced by Golay in 1958 for gas chromatographic separations. Since then, these columns have largely replaced packed columns in gas chromatography due to their superior efficiency.

 

Theoretical studies have long suggested that narrow open tubular columns could provide exceptional separation efficiencies in liquid chromatography. However, this potential remained unrealized until recently when Professor Liu successfully reduced the column diameter to approximately 2 micrometers—just one-fiftieth to one-hundredth the thickness of a human hair—paving the way for groundbreaking advancements in liquid chromatography.

 

Professor Liu's team has transformed this decades-old high-efficiency separation concept into reality by overcoming key technical challenges. They developed a method for fabricating ultra-narrow open tubular columns, optimized separation procedures, and, most notably, achieved record-high separation efficiencies. Their work represents the leading edge of this field.

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ETBR’s product line will include NOTLC instruments, columns, reagent kits, training, and services.