Abstract
Analytical methods founded upon whole cell-based assays are of importance in early stage drug development and in fundamental studies of biomolecular recognition. Here we have studied the binding of the monoclonal antibody trastuzumab to human epidermal growth factor receptor 2 (HER2) on human ovary adenocarcinoma epithelial cancer cells (SKOV3) using quartz crystal microbalance (QCM) technology. An optimized procedure for immobilizing the cells on the chip surface was established with respect to fixation procedure and seeding density. Trastuzumab binding to the cell decorated sensor surface was studied, revealing a mean dissociation constant, KD, value of 7 ± 1 nM (standard error of the mean). This study provides a new perspective on the affinity of the antibody-receptor complex presented a more natural context compared to purified receptors. These results demonstrate the potential for using whole cell-based QCM assay in drug development, the screening of HER2 selective antibody-based drug candidates, and for the study of biomolecular recognition. This real time, label free approach for studying interactions with target receptors present in their natural environment afforded sensitive and detailed kinetic information about the binding of the analyte to the target.
Highlights
Cell-based assays are interesting for drug development and diagnostics
We have addressed the use of the human ovary adenocarcinoma epithelial cancer (SKOV3) cell line, with an overexpression of the breast cancer-related human epidermal growth factor receptor 2 (HER2), as a means for studying the interaction with the monoclonal antibody-based drug trastuzumab, and with the long term goal of establishing methods for the rapid screening of new antibody-based candidate drugs
In this work we have studied the binding of the monoclonal antibody trastuzumab to the receptor HER2 on SKOV3 epithelial cancer cells using quartz crystal microbalance studies (Figure 1)
Summary
Cell-based assays are interesting for drug development and diagnostics. Even though the standard methods for early drug development are based on in vitro-assays, cell-based assays offer extended possibilities for understanding the biology including in vivo-effects [1,2]. The initial development of applications based upon cells attached to QCM surfaces have included the monitoring of cell adhesion [9], the effects of anti-cancer treatments on cells [29,30], the detection of cancer cells [31,32] and the affinity of antibodies for a cell membrane receptor [33]. In this study, developed a system for measuring the interaction to HER2 on SKOV3 epithelial cancer cells attached to a COP-1. The SKOV3 cell line was attached and fixed to COP-1 chips and trastuzumab was passed over the surface allowing binding to the HER2 on the cell membrane
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
