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Study of the Interaction of Trastuzumab and SKOV3 Epithelial Cancer Cells Using a Quartz Crystal Microbalance Sensor
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences. (BBCG)
Attana AB.
Attana AB.
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences. Uppsala University. (Linnaeus Ctr Biomat Chem, BMC;BBCL)ORCID iD: 0000-0002-0407-6542
2015 (English)In: Sensors, ISSN 1424-8220, E-ISSN 1424-8220, Vol. 15, no 3, p. 5884-5894Article in journal (Refereed) Published
Abstract [en]

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, K-D, 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.

Place, publisher, year, edition, pages
MDPI , 2015. Vol. 15, no 3, p. 5884-5894
Keywords [en]
quartz crystal microbalance, breast cancer, cell-based biosensor, Herceptin, trastuzumab, HER2
National Category
Biochemistry and Molecular Biology
Research subject
Natural Science, Biomedical Sciences
Identifiers
URN: urn:nbn:se:lnu:diva-37513DOI: 10.3390/s150305884ISI: 000354160900066PubMedID: 25763651Scopus ID: 2-s2.0-84928680424OAI: oai:DiVA.org:lnu-37513DiVA, id: diva2:753091
Available from: 2014-10-07 Created: 2014-10-07 Last updated: 2018-11-16Bibliographically approved
In thesis
1. QCM-based sensing using biological and biomimetic interfaces
Open this publication in new window or tab >>QCM-based sensing using biological and biomimetic interfaces
2014 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

The objective of this thesis was to explore novel approaches for studying molecular recognition at biological and biomimetic surfaces using the quartz crystal microbalance (QCM) biosensor technique. The first two papers focused on the synthesis and study of biotin selective polymer films prepared using the molecularly imprinted polymer (MIP) technique. Control over polymer structure is of importance for sensor reproducibility and sensitivity, and was addressed in Paper I where a simple strategy for fabricating uniform thin biotin imprinted polymer films was employed. In Paper II the binding of biotin moieties to thin (3-5 nm) biomimetic polymer films was examined and consequences for sensor performance discussed. The potential for using QCM as a tool for assessing the binding of small peptides derived from phage display screening was presented Paper III. Here, screening of a phage peptide library against immobilized adenine resulted in candidate peptides that were studied using this technique. In Paper IV a whole cell-based biosensor was developed for studying interactions with cell membrane-incorporated targets. Epithelial cancer cells, SKOV3, were attached to QCM sensor chips and the binding of the monoclonal antibody HerceptinTM was studied. This approach demonstrates the potential of using QCM to study binding to membrane-incorporated targets, an alternative to assays based upon immobilized receptor structures lacking their natural context.

Place, publisher, year, edition, pages
Växjö: Linnaeus University Press, 2014
Series
Linnaeus University Dissertations ; 192/2014
Keywords
Quartz crystal microbalance, molecularly imprinted polymers, phage display, interactions, artificial receptors, recognition, selectivity
National Category
Chemical Sciences
Identifiers
urn:nbn:se:lnu:diva-37514 (URN)978-91-87925-20-7 (ISBN)
Public defence
2014-10-31, N2007, Smålandsgatan 26B, Kalmar, 09:30 (English)
Opponent
Supervisors
Available from: 2014-10-08 Created: 2014-10-07 Last updated: 2014-10-08Bibliographically approved

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Elmlund, LouiseNicholls, Ian A.

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