Collaborate 2 Cure
December 1, 2016

Location : MRIGlobal

425 Volker Blvd, Kansas City, MO 64110
Or join virtually via “Zoom”

Quest for Developing Novel Immunological Assays for Cancer Immunotherapy


As part of the Kansas Vaccine Institute, the mission of the Immunology Core Lab (ICL) is to be the primary provider of immunology related analysis for basic research and vaccine development in Kansas. In particular, we provide researchers with the expertise to successfully perform immunological experiments that help them advance their projects. The most unique characteristic of the ICL is its ability to adapt and modify its techniques in order to provide tools for research areas other than immunology and vaccinology. The current capabilities of the ICL make it a great asset for the challenges involved in C2C’s Immunotherapy for Cancer research.

Francisco Martinez Becerra , PhD

Dr. Francisco Martinez Becerra obtained his PhD from the National Autonomous University in Mexico (UNAM). He then held postdoctoral positions at the UNAM’s School of Medicine in Mexico, at the Center for Vaccine Development of the University of Maryland at Baltimore and in Oklahoma State University. He has worked in subjects ranging from H. pylori and gastric cancer to Shigella and Yersinia pestis vaccines. He is currently a Research Assistant Professor at KU-Lawrence, and is the director of the Kansas Vaccine Institute’s Immunology Core Lab.

Isolation and Molecular Profiling of Circulating Tumor Cells with Divergent Phenotypes


Liquid biopsies are becoming an attractive source of biomarkers. One of the principle biomarker related to epithelial cancers found in blood is circulating tumor cell (CTC) enumeration of which revealed prognostic value in cancer patients. The challenge associated with CTCs as biomarkers has been the modest clinical sensitivity and specificity demonstrated using the FDA-approved platform. The question arises: does the biology limit the CTC burden or is the analytical platform used for their isolation limiting? Indeed, many microfluidic platforms have shown higher clinical sensitivity compared to the FDA-approved test. CTCs expressing invasive phenotypes down-regulate epithelial antigens, such as the epithelial cell adhesion molecule – EpCAM, which is typically used for the affinity selection of CTCs. It is believe now that CTCs may have a continuum of phenotypes and thus, a single selection marker (i.e., EpCAM) may not address all cells comprising the tumor microenvironment. We have developed a CTC selection strategy that employs two polymeric microfluidic devices (Figure 1) modified with monoclonal antibodies and connected in series with each selecting a distinct CTC subpopulation from a single blood sample. In addition to the common marker used for CTC positive selection (EpCAM), Fibroblast Activation Protein alpha (FAPα) expressing CTCs were also selected. Using the dual selection strategy, both CTC types were detected from patients with clinical sensitivity that showed significant improvement compared to selection in which only EpCAM was used. Approximately 90% of the selected CTCs were found not to co-express both antigens. Owing to the high purity (>80%) and clinical yields of the dual selection strategy, molecular analysis of both EpCAM and FAPα CTCs could be carried out, including next generation sequencing. Our results suggest FAPα and EpCAM CTCs can be used in concert to monitor disease progression or potentially guide therapeutic decisions. In this presentation, I will discuss the microfluidic chips we use for the selection of CTCs, the clinical results secured using these chips and the molecular profiling of these rare cells.

Malgorzata A. Witek, PhD

Dr. Maggie A. Witek received MSc. and Engineering degree from the Department of Chemistry and Chemical Technology at Silesian University of Technology in Gliwice, Poland in 1997. She earned Ph.D. in Analytical Chemistry from Michigan State University in 2002. She joined Prof. Steven A. Soper research group in the fall of 2002 in the Department of Chemistry at    Louisiana State University. In 2004 Dr. Witek joined the Center of BioModular Multiscale Systems (CBM2) at LSU as a Research Associate. In 2011 Dr. Witek joined Department of Biomedical Engineering at the University of North Carolina in Chapel Hill as an Assistant Research Professor. The main focus of her work at UNC was collaboration with oncologists at the UNC School of Medicine and development of the assay for isolation of low abundant subpopulations of circulating tumor cells from clinical samples using antibody modified microfluidic chips. Dr. Witek is currently Associate Research Professor in the Department of Chemistry at the University of Kansas in Lawrence. Her research will involve microfluidic-based isolation of liquid biopsy markers, including CTC followed by molecular profiling/sequencing of these cells; and selection of exosomes and cell free DNA for cancer and stroke diagnostics. She continues her work in the Center of BioModular Multiscale Systems for Precision Medicine as an investigator and coordinator of the Center’s Collaborative and Service Projects.