Breaking the ICE

LINCOLN, Neb.—LI-COR Biosciences is working with MitoSciencesto create specific assays used for disease and metabolic pathway research undera collaboration agreement the companies announced last month.

MitoSciences recently released three In-Cell ELISA (ICE)assays that monitor the stress-induced, species-variant, or tissue-specificchanges or differences in the levels of mitochondrial and other metabolictargets. These assays, also known as In-Cell Western (ICW) Assays, offer ahigh-throughput method to determine protein levels in cultured cells that isboth reproducible and quantitative, LI-COR claims in its announcement.

"Our drug development customers are increasingly interestedin using ICE assays to understand the extent to which their drugs' beneficialor adverse effects are unique to specific species of organs," says MitoSciencesCEO John Audette. "Our assays are focused in the most general sense on thevarious mechanisms that regulate cellular homeostasis, a very broad category ofresearch areas that includes metabolism, cell death, oxidative stress and otherprocesses that touch many important diseases. We have found the In-Cell ELISAplatform to be very well-suited to analysis of such systems, being able to'freeze' cells in their native states prior to quantitative measurement of keyprotein variables."

MitoSciences' drug-developer customers are excited about theIn-Cell ELISA platform, and the companies are undertaking collaborations toextend the assays to novel primary cell model systems, Audette adds.

"Basic researchers are also starting to appreciate the valueof the platform with its simplicity and quantitative power. For any researcherworking on cell-based analyses, ICE offers compelling advantages over Westernblotting," he adds.

LI-COR and MitoSciences came together at the time MitoSciencesreleased its first assay, says Jim Wiley, LI-COR's senior strategic marketingmanager.

"Researchers are able to obtain quantitative, highlysensitive data when using tools like near-infrared dye technology," Wileyclaims. "Scientists benefit from both companies' technologies when they imagetheir infrared ICE assays from MitoSciences on a LI-COR Odyssey InfraredImaging System," which the company claims sets the standard for quantitativenear-infrared fluorescence imaging. LI-COR also manufactures IRDye fluorescentdye-labeled secondary antibodies that are incorporated into the MitoSciencesICE kits.

"MitoSciences has proven that LI-COR's technology providesan excellent platform for our antibodies, both in kits specific to particularresearch problems, as well as in broad screening approaches used for evaluatingpathway-level changes due to diseases or drug effects," says Rod Capaldi, chiefscientific officer of MitoSciences. "We are already seeing adoption of the kitsfor drug screening efforts, and are also undergoing collaborations with leadingdrug developers to demonstrate the technology's potential as a broad screeningapproach."

"LI-COR customers, who are very familiar with the benefitsthat infrared fluorescent imaging provides, can now image these new, novelinfrared fluorescent-based assays, developed for specific, targeted pathwaysfrom MitoSciences on the Odyssey platform," Wiley notes. He adds thatMitoSciences also provides fully validated antibodies for targeted pathways ofwhich Odyssey customers can now take advantage. When an antibody is used forresearch, it is important to know that the antibody is reacting only with thetarget enzyme. In using antibodies for ELISA, the antibody must capture onlythe target enzyme or a complex containing that protein.

MitoSciences points out that many antibody developers limittheir validation to demonstrating that an antibody binds to orimmunoprecipitates a peptide or an over-expressed protein. This limited levelof analysis gives little, if any, confidence that the antibody can actuallycapture or label a native enzyme in a complex cell or tissue sample, or thatits affinity is limited to the desired target. MitoSciences' validation processinvolves a double immuno-precipitation followed by mass spectrometry analysisduring the development process. The company also uses cell and organ tissuesamples for screening at every stage in development, ensuring that theantibodies will work with the samples that matter to their research customers.

"The ability to use these assays on the Odyssey family ofimaging platforms will combine to be another tool for advancing cancerresearch," Wiley concludes.