New technology enables single-cell sorting based on functional outputs

For nearly 40 years, drugmakers have used genetically modified cells as mini drug factories. These cells can be programmed to secrete compounds that produce drugs used to treat cancer and autoimmune diseases such as arthritis.

Efforts to develop and manufacture novel biological therapies may benefit from a new technique for rapidly sorting single living cells in a standard laboratory setting. Using microscopic bowl-shaped hydrogel containers called “nanophils,” a UCLA-led research team recently demonstrated the ability to select cells based on their type and compounds -; And how many of these compounds -; excrete. The study was published in the journal ACS nano.

Technology can also advance basic biological research.

With this technology, the scientific community can reveal new insights into the key biological processes that represent a large part of our protein-making genes. I think the single cell is the quantum limit for biology. A nanofil is the evolution of a petri dish to the basic limit of a single cell.”

Dino DiCarlo, corresponding study author and Armond and Elena Hayrapiti University Professors of Engineering and Medicine, UCLA Samueli School of Engineering

DiCarlo, who is also a member of the California NanoSystems Institute at UCLA and the Johnson Comprehensive Cancer Center at UCLA, said the use of nanomotors is helping scientists overcome the limitations of other tools for measuring cell secretions.

The most common technique uses a network of small plastic containers called a microwell plate, but this approach lacks the nanofilament’s ability to sort single cells, and current technology typically requires weeks to grow enough cells so that exudates can be detected. Another alternative is a multi-million dollar instrument, found in a few dozen laboratories around the world, that measures secretions of about 10,000 cells in an experiment and can sort out live cells.

Compared to that tool, nanofilm can be used for much larger assays -; in millions of cells -; at a fraction of the relative cost -; Less than 1 cent per cell, for $1 or more per cell using the current standard.

Nanocytes are so small that it would take 20 million to fill a teaspoon with them. Specific to capture specific types of cells, they can be augmented with molecules that bind to cell secretions and glow under colored light. Because nanovials are made of hydrogel -; A polymer that holds almost 20 times its mass in water -; They provide a moist environment relatively similar to the natural homes of cells.

In the study, the researchers examined cells that were engineered to secrete a specific antibody drug. Using nanophils and a common analytical device called a flow cytometer, they selected cells that secreted the most of this antibody, and then grew those cells into colonies that produced 25% more of the drug than colonies that were not specifically selected.

The ability to produce antibody drugs with such increased efficiency could reduce the cost of drug production by a similar percentage, DiCarlo said.

The scientists also showed that they could pick out rare antibody-secreting cells that specifically bind to a target molecule, and they could pinpoint the DNA sequence information of the secreted antibody. This trial, an essential part of discovering new antibody drugs, took a day. Traditional methods may take weeks.

Researchers are currently using nanocells to study immune cells called T cells, which are used in cellular therapies, as well as to explore poorly understood biological phenomena. Nanovial technology is also the basis for a startup company, Partillion Bioscience, based on the UCLA campus in CNSI’s Magnify incubator.

Joseph de Root, co-first author on the paper, who received his Ph.D. from UCLA in 2020 and is co-founder and chair of Partillion.

Robert DiMatio, who received a Ph.D. from UCLA in 2021, is the paper’s other co-first author. Other UCLA co-authors are Manny Archang, Soohyung Lee, and Kyung Ha, who recently received their Ph.D. current doctoral students Mark Van Zee, Doyun Koo, Michael Melody and Shreya Udani; Associate Project Scientist Alison Scharow; James Eichenbaum, recently BA; and Professors Andrea Bertozzi and Robert Damoiso. Other authors are from Johns Hopkins University and the University of Houston.

The study was supported by the National Institutes of Health, the Stem Cell Research Fund in Maryland, and the Simmons Foundation.

source:

California Nano Systems Institute at the University of California

Journal reference:

de Root, c. et al. (2022) Suspensionable nano-hydrogel for parallel, single-cell functional analysis and sorting. ACS Nano. doi.org/10.1021/acsnano.1c11420.

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