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Canopy Biosciences Using Zellkraftwerk Buy to Move Into Protein Analysis

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NEW YORK — With its April purchase of German cytometry firm Zellkraftwerk, Canopy Biosciences is moving into the protein analysis space.

The St. Louis, Missouri-based firm has traditionally focused on tools for genomic research, offering technologies including CRISPR products and next-generation sequencing error correction developed by researchers at Washington University in St. Louis. It also offers protein analysis services using Meso Scale Discovery's platform, but this is typically used by customers investigating single proteins of interest, said Edward Weinstein, Canopy's co-founder, president, and CEO.

The Zellkraftwerk acquisition gives the company (which did not disclose the purchase price) a single-cell proteomic technology complementing its genomic offerings, Weinstein said.

Headquartered in Leipzig, Germany, Zellkraftwerk last month received a pair of German innovation awards for its ChipCytometry technology, which works by fixing and immobilizing target cells in microfluidic chips where they are stained with an antibody to a protein marker of interest and analyzed using automated epifluorescence microscopy.

The sample can then be photobleached and stained with another antibody to allow for multiplexed biomarker detection.

According to Weinstein, the company has currently validated 110 antibodies for use with the system, and, in theory, researchers could multiplex the full set, allowing them to measure 110 proteins per sample. Typically, however, the company's customers are looking to measure in the range of 25 to 40 proteins per sample.

A variety of single-cell protein imaging technologies have emerged in recent years, including systems like Fluidigm's mass cytometry-based Hyperion, IONPath's multiplexed ion beam imaging instrument, and Akoya Biosciences' CODEX system.

Zellkraftwerk's ChipCytometry platform similarly offers allows for multiplexed protein imaging but with the added ability to store and reanalyze samples down the road.

"You can run your sample on the chip and then you can stick it in the refrigerator for two years," said Thomas Campbell, product manager at Canopy, adding that this is particularly attractive for customers doing clinical research.

"You may just want to look at a few biomarkers initially, but then down the line you realize that you may wish that you would have looked at some additional markers," he said. "So you're able to delay the need to make a decision about which biomarkers you have to look at, which is really important for a clinical study that can take a number of years."

The ability to store and reanalyze is also potentially useful for researchers using rare samples, allowing them to experiment with different marker sets on the same samples.

Canopy was also drawn to the technology by its high spatial resolution and dynamic range. The platform offers resolution of 500 nanometers/pixel and eight orders of dynamic range, Campbell said.

"That's especially important for tissue applications where you have to get really high-resolution images in order to get single-cell data so you can have quantitative [protein] expression levels for each individual cell in a tissue sample," he said.

Weinstein said that Canopy offers the ChipCytometry platform both as an in-house service and as a product for purchase by researchers. He noted that, as is the case for most single-cell protein imaging systems, immune profiling is a major area of focus for the technology.

"Immune profiling is where this becomes critical," he said. "If you want to look at immune cell activation, you can scan with 25 or 30 markers across B cells, T cells, macrophages, just about everything you can think of. And not only in PBMCs, but you can do that in tissue, as well."

"If you look at our panel of markers, the vast majority of them have to do with looking at proliferation or immune response," he said. He added that the device has seen uptake within biotech and pharma as well as among academic researchers.

The company offers two sizes of chips, one with a capacity of around 250,000 cells and the other with a capacity of around 1 million. The system's throughput depends on the number of cells and number of markers a customer wants to analyze, but Weinstein said that a project analyzing 20 proteins in 10 different samples would take one to two days. Throughput is inherently limited by the technology's iterative staining approach, with each round of staining limited to around four or five markers, but an automated system, called CytoBot, is available for users running higher volumes of samples. The system uses Tecan hardware to automate the full ChipCytometry workflow from sample processing through to scanning and photobleaching and can be linked to up to three scanners.

Canopy typically does not develop new technologies itself but, rather, licenses or acquires technologies that have already been developed and validated and then modifies them as necessary for its customers, Weinstein said. In the case of the ChipCytometry platform, the firm is currently working to add to its library of antibodies and to extend the system to the analysis of formalin-fixed paraffin-embedded tissues, which he said was largely a matter of validating the antibodies to work in this sample type.

Weinstein said the company has found that its customers are typically interested in studying their samples using multiple omics tools.

"Researchers who are interested in understanding their samples using, say, RNA expression, using NanoString or RNA-seq [both offered by Canopy], are the same researchers who have needs around protein biomarkers," he said. "So when we have a multi-omic offering where researchers can kind of pick things a la carte, we find that they tend to be interested in multiple omic technologies."

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