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Tymora Awarded NIH Grant to Advance Protein Phosphorylation Tech

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NEW YORK (GenomeWeb) – Startup Tymora Analytical Operations this month was awarded a Phase II SBIR grant from the National Institutes of Health to develop multiplexed and reverse-phase array versions of its pImago system for non-antibody-based detection of protein phosphorylation.

The two-year grant, which is worth $451,538 in its first year, will also be used to further develop the technology for high-throughput analysis of signaling pathways, with an eye toward applications in cancer drug development and treatment, according to Tymora President and CTO Anton Iliuk.

Protein phosphorylation is a post-transcriptional protein modification involved in a range of cellular activities, while abnormal protein phosphorylation has been associated with a range of disorders including inflammatory diseases and cancer.

Measuring protein phosphorylation generally involves the use of phospho-specific antibodies. However, these are expensive and time-consuming to produce, Tymora CSO Andy Tao told GenomeWeb. And because of the sheer number of phosphorylation sites within organisms, many aren't covered by existing antibody libraries, particularly for model organisms.

To address this issue, Tao, also a professor at Purdue University, and Iliuk, a postdoc in his lab developed pImago. The technology is essentially a protein phosphorylation-detection reagent based on water-soluble, globular nanopolymers that are multi-functionalized with titanium ions, which are known to selectively bind to phosphorylated residues, and biotin for detection.

Since founding Tymora in 2012, the researchers have published a number of papers on pImago-based detection of individual phosphoproteins on Western blot and ELISA formats, and the company currently offers kits for both methods in the US through its website via partnerships with Sigma-Aldrich and Expedeon, as well as internationally via a network of distributors.

More recently, Iliuk and Tao published data demonstrating pImago's ability to sensitively and quantitatively measure the total phosphoprotein levels of multiple cell lysates and samples from human cell lines.

With the NIH funding, Tymora is now aiming to further develop the technology for multiplex detection of phosphorylation in both antibody microarray and functional reversed-phase array formats.

For multiplexed applications, pImago "can be combined with regular antibody detection to measure both total protein expression and phosphorylation at the same time," Tao said. For reverse-phase array systems, the technology would replace phospho-specific detection antibodies. Once bound to phosphorylated residues, he added, the pImago reagent can be detected by conventional antibodies.

Tymora also sees drug discovery and clinical potential for pImago, Iliuk added. To that end, the firm is working to optimize the technology's use in the analysis of signaling pathways, an application where "it can be utilized for discovery of cancer targets, better tailoring of drugs to patients most likely to respond, and monitoring of disease progression during therapy," he wrote in an email.

While analyzing gene mutations can be used to identify particular cancer types, "there are often hundreds or thousands mutations present, most of which do not progress cancer," Iliuk explained. "Many more can appear over time, especially with therapy treatment — a common cause of relapse."

By instead focusing on the protein phosphorylation changes triggered by these mutations, it may be possible to identify pathways involved in cancer progression and select the appropriate therapies for specific patients, he added.

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