Bioscale this week presented data on the application of its acoustics-based protein detection technology to measure and quantitate the activity state of multiple kinases in the MAPK pathway.
Researchers from the company shared the proof-of-concept data in a poster at the American Association for Cancer Research annual meeting, demonstrating that the firm's acoustic membrane microparticle, or AMMP, assays can detect a range of phosphoproteins with greater sensitivity and a simpler workflow than current methods.
The presentation comes as Bioscale — founded in 2002 by CEO Mark Lundstrom — seeks to build a collection of research to robustly demonstrate both the sensitivity and the potential flexibility of its ViBE protein analytics system.
Bioscale's ViBE system relies on the company's AMMP technology, an acoustic, as opposed to optical, detection system. A capture agent attached to a resonating membrane is exposed to a sample containing the analyte of interest. The analyte is captured by an affinity agent, which adds mass to the membrane and changes the frequency of its oscillations. The system detects and quantitates the target analyte by measuring these frequency changes (PM 6/18/2010).
Aberrant activity of MAP and PI3 kinases has been implicated in a number of cancers, making measurement of these pathways key to cancer drug development efforts. While Bioscale declined to provide details on the activities of specific customers, BioScale COO Chip Leveille told ProteoMonitor this week that adopters of the AMMP technology are indeed pursuing research along these lines.
In the proof-of-concept studies of AMMP pathway analysis presented this week, the group created six separate assays measuring multiple kinases including EGFR, MEK, ERK, AKT, p38, and JNK.
"Our goal was to be able to use our technology to clearly represent that we could quantify multiple different kinases as a single assay [plate]," Matthew Dickerson, the Bioscale senior scientist who presented the results at AACR, told ProteoMonitor.
The researchers compared lysates from multiple unstimulated tumor cell lines to those from cell lines stimulated with ligands to several well-known surface receptors to measure expressed changes in their phosphorylation states.
The group demonstrated that the technology could measure multiple phosphoproteins from a portion of the MAPK pathway on a single plate, allowing the analysis of groups of analytes at the same time under the same conditions. Using the system, they demonstrated that "EGF stimulation causes the phosphorylation [of] the EGF receptor, ERK, MEK and p38 in A431 cells and ERK, MEK, and p38 in MCF-7 cells," the group reported in the poster.
The researchers also developed assays to monitor kinase dimerization between MEK and ERK by rearranging the pairing of antibodies from the activity assays, which, they said, highlighted the ability of the technology to detect weak, transient, low-affinity interactions.
"The one thing that everyone is interested in is sensitivity — how little cell lysate or tumor lysate you need to do the analysis," Dickerson said. "We illustrated [in the poster] how our sensitivity lines up with a normal Western blot analysis, and throughout [the studies] we were able to use a lot less sample."
Overall, the group was able to measure multiple kinases in their native states in a single AMMP assay plate using fewer than 2,000 cells per assessment.
"For the EGF receptor assay, as few as several hundred cells were used for the analysis, while maintaining significant signal-to-noise ratios at the cell numbers tested," the group reported.
Dickerson said the results represent sensitivity "far superior to pretty much any technology available currently," such as ELISAs or systems that rely on optical detection.
The results are relevant most obviously to oncology, he noted, where a great deal of academic and pharmaceutical research is working toward kinase inhibition.
Leveille said the company has been collaborating with such groups, but noted that Bioscale's main focus has been on reiterating its technology's basic potential in studies like the one presented.
"[AMMP] operates on a fundamentally different level than ELISA-like technology or competitors you see out there that use chemoluminescent or other technologies," he said. "It's more directed in its measurement technique and, more importantly, it actually has the ability to measure these interactions more in their natural state and does not require the washes or harsh sample prep techniques you see in other technologies."
He added that the technology is able to study the "lowly expressed, highly transient interactions, which are really the mechanisms driving the signaling pathways and, quite frankly, more informative overall to what is actually happening and eventually what people will target for drug development."
According to Leveille, Bioscale is also working on demonstrating other panel types "with very specific markers looking at disease states [that] could be highly predictive but require multiple markers to actually be developed on one platform.
"The same principles apply to that, though that’s more plasma serum-based than looking at cell lysate or tumors," he said.
Though the company has said the technology could be a good fit for diagnostic applications, specifically for a potential point-of-care instrument, Leveille said that Bioscale is "not there yet."
However, he said the firm is seeing interest from commercial and academic partners in the "pre-clinical to clinical" arena.
"We're getting pulled quickly into that direction because ... it really doesn’t matter what model system we're in, we can generate the same results. The normal transition between different model systems requires revalidation. There is a lot less of that with our platform, so things are more comparative."
"It's all a lot easier, and so the ability to move more quickly to the clinic is evident," he said. "But we [want to] ensure that the science is right before we fully declare that that's where we're going."
Leveille said he could not provide details on the company's progress gaining customers and partnerships. "What we can say is that the number of customer interactions is growing rapidly," he said.
Novartis presented data earlier this year at the Society for Laboratory Automation and Equipment conference from its use of the AMMP technology to select well-expressed clones in protein production.
Leveille suggested that this presentation is only part of what Novartis is doing using AMMP. "I can't speak for them, but they know what the system is capable of, and I believe they have plans to take full advantage of that," he said.
Early last year Millennium Pharmaceuticals announced it had adopted Bioscale's ViBE system as part of its drug development workflow, using it to quantitate pharmacodynamic biomarkers as an alternative to AlphaScreens and LI-COR Western blot assays (PM 1/14/2011).
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