A novel approach to determining the affinity of protein–carbohydrate interactions employing adherent cancer cells grown on a biosensor surface

Peiris, Diluka; Markiv, Anatoliy; Curley, G. Paul; Dwek, Miriam

doi:10.1016/j.bios.2012.02.037

Cited by 34 publications

(34 citation statements)

References 30 publications

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“…The first immunosensor based on piezoelectric detection was reported in 1972 by Shons and co-workers [ 18 , 19 , 20 ]. The QCM technique has since been undergone significant development, whereby today it is possible to use synthetic polymer-based antibody mimics [ 21 , 22 ] and even cells as sensor recognition elements [ 7 , 23 , 24 , 25 , 26 , 27 , 28 ].…”

Section: Introductionmentioning

confidence: 99%

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Study of the Interaction of Trastuzumab and SKOV3 Epithelial Cancer Cells Using a Quartz Crystal Microbalance Sensor

Elmlund

Käck

Aastrup

et al. 2015

Sensors

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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.

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Section: Introductionmentioning

confidence: 99%

“…The possibility of studying the binding of an analyte to cells attached to resonator surfaces allows for the interaction to be examined in a more natural environment, i.e. , together with other cell membrane components that may affect the binding properties [ 7 , 23 , 24 , 25 ].…”

Section: Introductionmentioning

confidence: 99%

Study of the Interaction of Trastuzumab and SKOV3 Epithelial Cancer Cells Using a Quartz Crystal Microbalance Sensor

Elmlund

Käck

Aastrup

et al. 2015

Sensors

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“…Abnormal changes in the carbohydrate composition of cancer cell surface have been associated with the survival, invasion and metastasis of cancerous cells 2 . For instance, the metastatic colorectal cancer cells have an elevation in fucosylation in comparison to non-metastatic colorectal cancer cells 3 . Until now, many glycoproteins have been identified as biomarkers for various diseases, such as breast cancer and colorectal cancer 4 .…”

mentioning

confidence: 99%

Real-time and label-free analysis of binding thermodynamics of carbohydrate-protein interactions on unfixed cancer cell surfaces using a QCM biosensor

Song

Shuai

et al. 2015

Sci Rep

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A novel approach to the study of binding thermodynamics and kinetics of carbohydrate-protein interactions on unfixed cancer cell surfaces using a quartz crystal microbalance (QCM) biosensor was developed, in which binding events take place at the cell surface, more closely mimicking a biologically relevant environment. In this study, colon adenocarcinoma cells (KM-12) and ovary adenocarcinoma cells (SKOV-3) grew on the optimized polystyrene-coated biosensor chip without fixation. The association and dissociation between the cell surface carbohydrates and a range of lectins, including WGA, Con A, UEA-I, GS-II, PNA and SBA, were monitored in real time and without label for evaluation of cell surface glycosylation. Furthermore, the thermodynamic and kinetic parameters of the interaction between lectins and cell surface glycan were studied, providing detailed information about the interactions, such as the association rate constant, dissociation rate constant, affinity constant, as well as the changes of entropy, enthalpy and Gibbs free energy. This application provides an insight into the cell surface glycosylation and the complex molecular recognition on the intact cell surface, which may have impacts on disease diagnosis and drug discovery.

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“…In summary, the results clearly demonstrate that affinity estimated through

differs depending on whether EIS or QCM analysis is used for a lectin–cell binding model under Langmuir model premises. The capability of evaluating

as demonstrated herein can be helpful in tumor cell characterization and in the study of drug delivery [ 32 , 33 , 34 , 38 ].…”

Section: Resultsmentioning

confidence: 99%

“…QCM and EIS techniques have also been used to characterize lectin–cell interactions, including cell-surface glycosylation [ 32 , 33 ] or discrimination among malignant stages of tumor cells [ 19 , 32 , 33 , 34 ]. In this study, we performed the first comparative evaluation of the sensitivity between piezoelectric (by QCM) and impedimetric (by EIS) approaches in determining lectin–cell interactions.…”

Section: Introductionmentioning

confidence: 99%

Evaluating the Equilibrium Association Constant between ArtinM Lectin and Myeloid Leukemia Cells by Impedimetric and Piezoelectric Label Free Approaches

et al. 2014

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Label-free methods for evaluating lectin–cell binding have been developed to determine the lectin–carbohydrate interactions in the context of cell-surface oligosaccharides. In the present study, mass loading and electrochemical transducer signals were compared to characterize the interaction between lectin and cellular membranes by measuring the equilibrium association constant, Ka, between ArtinM lectin and the carbohydrate sites of NB4 leukemia cells. By functionalizing sensor interfaces with ArtinM, it was possible to determine Ka over a range of leukemia cell concentrations to construct analytical curves from impedimetric and/or mass-associated frequency shifts with analytical signals following a Langmuir pattern. Using the Langmuir isotherm-binding model, the Ka obtained were (8.9 ± 1.0) × 10−5 mL/cell and (1.05 ± 0.09) × 10−6 mL/cell with the electrochemical impedance spectroscopy (EIS) and quartz crystal microbalance (QCM) methods, respectively. The observed differences were attributed to the intrinsic characteristic sensitivity of each method in following Langmuir isotherm premises.

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A novel approach to determining the affinity of protein–carbohydrate interactions employing adherent cancer cells grown on a biosensor surface

Cited by 34 publications

References 30 publications

Study of the Interaction of Trastuzumab and SKOV3 Epithelial Cancer Cells Using a Quartz Crystal Microbalance Sensor

Study of the Interaction of Trastuzumab and SKOV3 Epithelial Cancer Cells Using a Quartz Crystal Microbalance Sensor

Real-time and label-free analysis of binding thermodynamics of carbohydrate-protein interactions on unfixed cancer cell surfaces using a QCM biosensor

Evaluating the Equilibrium Association Constant between ArtinM Lectin and Myeloid Leukemia Cells by Impedimetric and Piezoelectric Label Free Approaches

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