Surface plasmon resonance-biosensor detects the diversity of responses against epidermal growth factor in various carcinoma cell lines

Abstract

Surface plasmon resonance (SPR) biosensor detects intracellular signaling events as a change of the angle of resonance (AR). We previously reported that the activation of epidermal growth factor receptor (EGFR) on keratinocytes causes a unique triphasic change of AR, whereas the activation of other receptors, such as IgE receptor and adenosine A3 receptor on mast cells, causes a transient monophasic increase of AR. To study the mechanism of AR changes induced by EGFR activation, we introduced wild and mutated EGFR cDNAs into Chinese hamster ovary (CHO) cells and analyzed changes of AR in response to EGF. CHO cells expressing wild-type EGFR showed a triphasic change of AR, whereas cells expressing kinase-dead EGFR (K721M) showed minimum change of AR. A phosphatidylinositol 3-kinase inhibitor, wortmannin, attenuated the third phase of AR change in CHO cells expressing wild-type EGFR. The pattern of AR change was independent on the concentration of EGF. We also analyzed changes of AR with a nontumorigenic keratinocyte cell line, HaCaT, and several cell lines of carcinoma to explore the feasibility of SPR biosensor as a tool for clinical diagnosis. The activation of HaCaT cells and one out of six carcinoma cell lines showed a full triphasic change of AR. In contrast, five out of the six cell lines showed mono- or bi-phasic change of AR. These results suggest that EGF induces the SPR signals via the phosphorylation of EGFR, and provide a possibility that the SPR biosensor could be applied to the real-time detection and diagnosis of malignant tumors.

Surface Plasmon Resonance: A Versatile Technique for Biosensor Applications

“…Next, solutions containing stimuli are flowed over the cells, and their response is measured in real time by SPR. This strategy has been successfully applied to detect and diagnose malignant tumors cells, such as Chinese hamster ovary cells without the use of immunological labels [ 86 ]. Liu et al designed an SPR biosensing system with anti-VEGF antibodies immobilized on the surface and living SKOV-3 ovarian cancer cells cultured on the flow cell chamber ceiling [ 87 ].…”

Surface Plasmon Resonance for Cell-Based Clinical Diagnosis

“…Chen et al demonstrated that GPCR-mediated SPR responses in CHO cells were induced by density changes on the sensor chip surface [15]. Finally, the inhibition of receptor activities by molecular engineering totally abolished SPR signals, preserving the binding activities of ligands [14,16,17]. These observations demonstrate that RI near the plasma membrane, which might reflect accumulation and rearrangement of proteins activated by intracellular signal transduction, dramatically changes in response to exogenous stimuli.…”

Angle-Scanning Surface Plasmon Resonance System with 3D Printed Components for Biorecognition Investigation

“…Surface plasmon resonance (SPR) is a powerful analytical technology. It can detect the thickness of the films absorbed onto the sensor surface and interactions between biomolecules such as antigen-antibody or protein-DNA [1][2][3][4][5]. In comparison with traditional detection methods, like X-ray, HPLC, and HPLC-MS, SPR technology can perform kinetics parameters (association and dissociation process) measurement, concentration measurement, and real-time molecule detection.…”