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CyBio and Axxam Partner on Alternative Photoprotein for Luminescence Assays

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German instrumentation maker CyBio and Italian drug-discovery services firm Axxam announced last week that they have signed an agreement to develop and market flash luminescence-based cellular assays for drug discovery.

The collaboration will pair the CyBi Cellight, CyBio’s recently overhauled automated luminescence plate reader and liquid handler, with Photina, a new photoprotein from Axxam for conducting cell-based flash luminescence assays.

Further financial details of the partnership were not disclosed. However, Germano Carganico, Axxam’s chief business officer, said last week that Axxam will grant CyBio non-exclusive access to the Photina technology and “cell biology know-how,” and in return, CyBio will provide Axxam with an undisclosed number of CyBi Cellight instruments for use in its in-house screening services.

The “gold standard” photoprotein for luminescence assays is aequorin, derived from the jellyfish Aequorin aequorin. The intellectual property for aequorin is owned by Belgian drug-discovery firm Euroscreen.

According to Christoph Wenger, a product manager for CyBio, aequorin has been the most commonly used assay technology on most luminometers, including CyBio’s. But research has shown that the Photina protein is brighter and more stable; thus, CyBio found it advantageous to validate it on the CyBi Cellight.

“It’s an important assay to do functional testing of GPCRs, and also calcium channels,” Wenger said. “These are two target groups that are very important in drug discovery, and it covers a lot of drug research being done today. This is a sophisticated but straightforward assay for these targets.”

Wenger also said that it had an official agreement in place to license aequorin from Euroscreen, as well, and that the Axxam agreement would not change that. “It’s just another assay that can be run on our instrument,” Wengert said.

For CyBio, the deal may also mark an opportunity for the Jena-based company to make some noise in the market for high-throughput assays based on intracellular calcium release, such as those for GPCRs and ion channels — a market that instrumentation giant Molecular Devices has a stronghold on thanks to its FLIPR line of plate readers.

According to Molecular Devices product manager Jennifer McKie, the multi-purpose FLIPR instruments, which include FLIPR3 and the recently introduced FLIPR Tetra, have flash luminescence capabilities, but are more commonly used with fluorescent reagents, such as the Fluo-3 and Fluo-4 calcium indicators.

“It’s the same end result, but CyBio’s probably more sensitive than the FLIPR is [for luminescence],” McKie said. “It’s a market we’re more interested in getting into. It’s something that our customers are interested in, as being an alternative to a fluorescence assay. The FLIPR can do some brighter luminescence assays, like some of the brighter aequorin assays, or Photina, which is brighter than aequorin. So we’re definitely targeting the brighter luminescence market.”

Molecular Devices has worked with Axxam to test Photina on the FLIPR instruments. Axxam was one of the early-access partners for FLIPR Tetra, and when FLIPR Tetra was launched at Molecular Devices’ June 2004 user meeting, Axxam presented data obtained with Photina on FLIPR3 compared with FLIPR Tetra. According to company officials, however, Axxam and Molecular Devices have not signed an official agreement surrounding the two technologies as yet.

“The reason for having done some experiments together with Molecular Devices is that … FLIPR was not able to detect signals from aequorin, because it was too weak,” Carganico said about Axxam’s collaboration with MDCC. “With our photoprotein, we are able to develop flash luminescence assays which can be run on FLIPR.”

Wenger hopes that providing customers with the ability to use both aequorin and Photina on the CyBi Cellight will help it sell instruments to those seeking an alternative to fluorescence assays.

“The labeling of the dye in fluorescence assays is often toxic to the cells, if the cells are incubated with the dye for a longer time,” Wenger said. “Whereas the aequorin and the Photina are non-toxic, so they don’t change the physiology or pharmacology of the cells.

“Also, with fluorescence, there can be a high number of false positives due to auto-fluorescence,” Wenger added. “You have fewer artifacts with flash luminescence assays.”

Wrestling market share from Molecular Devices won’t be easy however, as the company has a strong market presence and a large sales force throughout both Europe and the US. Although both companies declined to provide specific sales figures, Wenger conceded that Molecular Devices has a strong reputation.

“Looking at the application, Molecular Devices is the biggest competitor,” Wenger said. “It’s a different type of assay. If people consider which assay and what instrument fit best to analyze a certain target, from that point of view, they are the biggest competitor.”

Cash Cow for Axxam?

Axxam’s stake in the deal comes from a slightly different angle. According to Carganico, Axxam is a drug-discovery service provider first and foremost, not a reagent provider.

“We mainly develop assays — this is the core business of the company,” Carganico said. “The goal is to develop new tools, as is the case with Photina. The reason for developing these, in the beginning, is not that we want to make money with these tools, in a sense.

“Our goal is to be discovery partners for our customers,” Carganico added. “You often find tools that can provide added value, and this is the case here. The project started with the aim of having the freedom to operate — to have a photoprotein that is proprietary, so we do not have to pay licenses to other companies.”

That is not to say that Axxam won’t welcome supplemental revenues that Photina might generate. “At the end of this project, it turned out that Photina was much better than the available photoproteins on the market,” Carganico said.

Axxam already has licensed Photina to four undisclosed companies in pharma and biotech, and according to Carganico, as many as ten companies are currently testing the assay technology.

“The only licensing so far is related to Photina, because in some cases, people are asking us to have the photoprotein itself, not linked to any particular assay development by us,” Carganico said. “They will typically ask for a cell line expressing Photina, or we may also provide the plasmids to them, to use in their cellular samples. Again, though, it’s not the main revenue source for us, it’s just added value.”

Another benefit Axxam hopes to gain from the CyBio deal is increased capability as a screening company. Currently, Axxam does mostly assay development for clients, with a small amount of screening that Carganico said was not high-throughput.

But because the Cellight platform is a complete high-throughput system — including the Lumax Flash HT luminescence reader, analysis software, and 1,536-well plate contact-free multi-channel reagent dispenser — Axxam hopes it can upgrade its screening service offerings.

“We’ve developed assays for HTS in a sense, because we work with 1,536-well technology, but we don’t have a scanning machine in house to screen a 2 million-compound library,” Carganico said. “We can go to about 20,000 test compounds a day. But this will change, and probably will change with the agreement with CyBio.”

Carganico said that it also hopes to explore collaborations — although not exclusive agreements — with other instrumentation providers besides CyBio and Molecular Devices.

Specifically, he said that PerkinElmer is “very much interested in the technology,” and that Tecan had also expressed interest. Calls to those companies were not returned in time for this publication.

“We don’t really have a formal agreement with Molecular Devices, and even the agreement CyBio is non-exclusive, because we don’t want to be linked to a specific technology provider,” Carganico said. “We really try to solve the problems of our partners with any technology available. I don’t know if this is the right strategy or not, but this is what we do — use the best tool and technology case by case.”

— BB

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