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Chromatography Spin-off Eprogen Hopes to Replace 2D Gels With HPLC Separation Method

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In an effort to attract investors to its multi-dimensional liquid chromatography-based separation technology, Eichrom Technologies last week spun off its proteomics operations as a wholly-owned subsidiary called Eprogen.

Initially, the new company will attempt to commercialize Eichrom’s ProteoSep separation technology with $3 to $4 million in technology and funds supplied by GCI, Eichrom’s parent company, Bruce Grotefund, CEO of Eichrom, told ProteoMonitor. But scaling up the technology, Grotefund added, will require more resources than GCI is able or willing to provide.

“It’s my feeling that in order to commercialize and get it to the market quickly it’s going to require some more horsepower,” he said. “So what we’re looking to do is raise some additional funding, preferably with a strategic investor; someone in the pharmaceutical or drug discovery area, or some type of organization that can provide funding and access to the market.

Eprogen’s ProteoSep technology, developed in collaboration with David Lubman, a researcher in the University of Michigan chemistry department, uses a two-dimensional liquid chromatography approach to separate whole proteins on the basis of isoelectric point, or pI, as well as by molecular weight.

In the first phase, Lubman has developed a method for separating large numbers of proteins on the basis of pI using commercially available Eichrom columns, Lubman said. Subsequently, the proteins in each of the pI fractions are separated by molecular weight using reverse-phase high-performance liquid chromatography (HPLC). These columns, packed with a proprietary non-porous, hard-sphere packing media, “provide a vast separation of large intact proteins that’s hard to do by regular HPLC,” Lubman said.

Prior to his collaboration with Eichrom, Lubman used 2D gel electrophoresis to separate proteins in his laboratory, but analyzing the gel spots using mass spectrometry was difficult and time-consuming, he said. After some preliminary work with Eichrom’s column packing material, “we got some really excellent results and decided that this was the way to go.”

The liquid-based separation method has other advantages over 2D gel electrophoresis, Lubman added. Because the separated proteins can be easily interfaced with a mass spectrometer, no spot picking instruments or a matrix are required. “We can get an exact molecular weight of intact proteins, or digest them for sequencing analysis,” he said. “With gels that’s hard to do because the proteins are embedded in the gel.”

Eprogen’s technology is similar to 2D gels in one respect, however. The accompanying software can graphically display the proteins in terms of pI versus molecular weight, much as they would in a 2D gel experiment. Lubman said this is important because it allows for easy visual comparison between distinct samples, and because a 2D gel-type display is “how biologists think of things.”

Other proteomics groups, including Celera Genomics and MDS Proteomics, have applied liquid chromatography-based methods to separating proteins, but have first used a tryptic digest to break the proteins into smaller peptide sequences. Lubman claims this approach may offer greater sensitivity, but presents a more complicated problem for researchers trying to determine the proteins that correspond to the peptide sequences.

However, the HPLC approach championed by Celera has proven to be a feasible technique for industrial-scale protein analysis, and offers significant advantages in resolution, said Scott Patterson, senior director for proteomics at Celera. “There may be some advantages to this method over others in specific situations, but it is yet to be proven,” he said.

Eichrom, with 13 employees, currently sells ProteoSep kits that contain software, protocols, and column packing materials for customers with liquid chromatography-based protein separation systems. Ultimately, Eprogen will attempt to commercialize an instrument based on the ProteoSep technology, Grotefund said, “but our first goal is to get out there and refine the sample anlaysis technique.”

Eprogen is beta testing the technology with several undisclosed partners in the pharmaceutical industry, and is in discussions with the National Institutes of Health and other academic laboratories. —JSM

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