Angiogenesis and the Search for Novel Biomarkers

The relationship between inflammation and cancer is an established one, but gaps remain in our understanding of how inflammation drives tumor initiation and progression. Angiogenesis, the process by which new blood vessels are formed, is a key area under investigation with the potential to fill these gaps. The increased production of chemokines, cytokines, proteolytic enzymes and growth factors associated with angiogenesis acts as a rich source of biomarkers allowing mechanistic insight and rationale behind the efficacy and toxicity of current therapies. These biomarkers also uncover targets for the development of novel anti-cancer therapeutics.

ELLA™: SENSITIVE AND SPECIFIC ASSAYS WITH A SIMPLE AND SPEEDY WORKFLOW

At Duke University, Andrew Nixon, Ph.D., M.B.A., an Associate Professor of Medicine, and Mark Starr, B.S., a Senior Research Analyst, are particularly interested in developing clinically relevant angiogenic biomarkers to guide innovative approaches for anti-cancer therapeutic intervention. They are spearheading an effort to develop robust assays for angiogenic factors that can produce high-quality data with limited sample volume. According to Dr. Nixon, “Sample volume needs and assay sensitivity are two of the biggest challenges we face when performing plate-based assays.” With Ella, he adds, “The lower volume usage allows for the expansion of research into novel areas of angiogenesis and inflammation as they pertain to many forms of cancer.” Patient samples are precious, often limited in volume and require smart planning to get more data points out of one assay run. Simple Plex™ assays on Ella are so sensitive that you can use as little as 2.5 µL of your sample. “Ella has allowed us to do more with less,” affirms Mr. Starr, who continued by saying, “Ella has allowed us to generate data with 50% less sample volume, compared with other standard plate-based methodologies.”

Traditional ELISA approaches are hands-on and commonly last the duration of a workday. With Ella, Dr. Nixon and his colleagues advanced to a simplified workflow that freed up valuable time for research. “The assay set-up time is very short. Simply dilute samples and run without the need for standard control preparations that add daily and user variability,” says Dr. Nixon. Indeed, Simple Plex assays on Ella require only 10 to 15 minutes of assay prep time. Once the run has started, you can come back to thoroughly analyzed data in 1 hour.

BIOMARKER DISCOVERY REQUIRES ASSAY FLEXIBILITY

While angiogenic and inflammatory marker analyses continue to be Dr. Nixon and Mr. Starr’s primary interest, they are also currently validating over 30 other biomarkers for large-scale clinical trial implementation. The scope of this work created the need to look for assay options with expanded capabilities. “Ella has validated assays that are interesting to us that other companies had not developed or where assays underperformed,” says Dr. Nixon. One example of this success is the development and clinical trial testing of the VEGF-C assay recently developed on the Ella platform. Alternate VEGF ligands have been implicated in predicting the efficacy of anti-angiogenic therapies in patients. Quality data focusing on VEGF-C levels have been lacking in the field. Currently, Dr. Nixon’s laboratory is testing VEGF-C using the 72x1 cartridge, which allows for up to 72 individual samples to be run in a single-analyte assay. When there is need to look at multiple targets simultaneously, their group can switch to the 16x4 cartridge, which enables multiplexing of up to four targets of interest for sixteen samples. In situations where assay availability and appropriate validation is limited, the Ella team has custom manufactured assays to facilitate further research progress. “ProteinSimple and the Ella system are great scientific tools to have in the lab. Their methods for assay validation are excellent, and the service is fantastic,” commends Dr. Nixon.