Customer Spotlight

A Simple Yet Powerful Method for Measuring Real-Time Kinase Activity

Customer Spotlight - AssayQuant Technologies, Inc.


Accelerating progress through innovation at the interface of Chemistry and BiologyAssayQuant Technologies, Inc., in Marlborough, Massachusetts is a growing Life Science company that develops innovative enzyme assay formats for a quantum improvement in performance and productivity to enable discovery or accelerate drug development. Its founders, Dr. Erik Schaefer and Dr. Barbara Imperiali, each impart a wealth of experience that allows AssayQuant to deliver exceptional value to their customers.

The company’s initial focus is on protein kinases, a diverse group of over 500 enzymes whose dysregulation lies at the center of many human diseases, including cancer. Indeed, more than 30% of all drug development efforts focus on protein kinases. Although more than 30 drugs have received FDA approval, and many more are in clinical trials, these are predominately ATP-competitive inhibitors.  These have several disadvantages, including cross-reactivity with other kinases or some of the many enzymes that use ATP and the fact that ATP is present at millimolar levels in our bodies. Recently, a surge in the generation of kinase inhibitors with different modes of action prompted a need for improved assay tools to effectively and efficiently characterize inhibitor mechanism of action and drive decisions earlier in the drug development process. Dr. Schaefer emphasizes "this is a huge challenge, where our technology is superbly positioned to drive productivity in drug development and efficacy that will benefit all who suffer from these debilitating diseases".

AssayQuant's mix-and-read PhosphoSens™ assay platform, is based on innovative technology covered by several patents exclusively licensed from the Massachusetts Institute of Technology (MIT). The platform uses optimized peptide substrates, that when phosphorylated in the presence of a kinase, ATP and Mg2+, uses chelation-enhanced fluorescence (CHEF) to generate a fluorescence signal. The result is a simple yet powerful method to measure protein kinase activity in a homogenous and continuous (kinetic) format using commonly available microplate readers. Their technology is ideal for elucidating drug mechanisms of action, and can be applied across the entire drug development process using a wide range of samples including purified enzymes and crude cell or tissue lysates. "These kinase sensors are sensitive enough to enable tissue diagnostics on a microscale, which may eventually allow them to serve as companion diagnostics", notes Dr. Schaefer.

An earlier version of the Sox technology was used by Dr. Schaefer to create the Omnia® Kinase Assay product line while at Invitrogen (now Thermo Fisher Scientific). As the current exclusive licensee of the Sox technology, Drs. Schaefer and Imperiali are actively integrating the most recent exciting improvements from the Imperiali laboratory at MIT and AssayQuant that result in superior substrates including higher reaction rates, tighter binding, lower backgrounds and increased selectivity. These next generation Sox-based substrates are the enabling content of the PhosphoSens™ platform.

Dr. Schaefer was familiar with BioTek’s washers and readers as part of his reagent development efforts at previous companies.  When he and Dr. Imperiali founded AssayQuant Technologies, they began looking for a microplate reader to use during PhosphoSens™ assay development. The team considered all the major microplate reader options, and also assessed trends in the research literature, where they found an increased frequency of BioTek readers.

Ultimately, Dr. Schaefer selected the Synergy™ Neo2 Multi-Mode Reader, citing its exceptional sensitivity and versatility for different plate densities, along with an integrated BioStack™ Microplate Stacker to increase automated throughput. Service and support were also critical factors, and as he recalls, “BioTek’s instruments are known for reliability and quality, and they also have a great reputation for support to make sure we are productive.” Indeed, in just a few months, AssayQuant has created assays for the first 135 targets, including both tyrosine and serine/threonine kinases. AssayQuant also offers very attractive Master Service Agreements to provide customers with new assays not currently in their current catalog offering. "We are very happy with our decision", says Dr. Schaefer. He also remarks that the Synergy Neo2’s multi-mode modularity aligns with the company’s longer term vision for new technologies and allows for benchmarking against existing technologies. As AssayQuant and its technologies continue to evolve, he is confident that Synergy Neo2 and BioTek Instruments can help him to deliver the best results to his customers.

Dr. Erik Schaefer discussing latest results with Susan Cornell-Kennon, Senior Research Scientist at AssayQuant

 Dr. Erik Schaefer discussing latest results with Susan Cornell-Kennon, Senior Research Scientist at AssayQuant, with results using PhosphoSens™ technology displayed in real time.


To learn more about AssayQuant Technologies, Inc., visit their web site.
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Thanks to Dr. Schaefer and Susan Cornell-Kennon at AssayQuant Technologies for sharing their BioTek experience.

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