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QCM-based sensing using biological and biomimetic interfaces
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
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
Keyword [en]
Quartz crystal microbalance, molecularly imprinted polymers, phage display, interactions, artificial receptors, recognition, selectivity
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:lnu:diva-37514ISBN: 978-91-87925-20-7 (print)OAI: oai:DiVA.org:lnu-37514DiVA: diva2:753103
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
List of papers
1. A Phage Display Screening Derived Peptide with Affinity for the Adeninyl Moiety
Open this publication in new window or tab >>A Phage Display Screening Derived Peptide with Affinity for the Adeninyl Moiety
2014 (English)In: Biosensors, ISSN 2079-6374, Vol. 4, no 2, 137-149 p.Article in journal (Refereed) Published
Abstract [en]

Phage display screening of a surface-immobilized adenine derivative led to the identification of a heptameric peptide with selectivity for adenine as demonstrated through quartz crystal microbalance (QCM) studies. The peptide demonstrated a concentration dependent affinity for an adeninyl moiety decorated surface (KD of 968 ± 53.3 μM), which highlights the power of piezoelectric sensing in the study of weak interactions. 

National Category
Biochemistry and Molecular Biology
Research subject
Chemistry, Biochemistry
Identifiers
urn:nbn:se:lnu:diva-37248 (URN)10.3390/bios4020137 (DOI)
Available from: 2014-09-24 Created: 2014-09-24 Last updated: 2017-02-16Bibliographically approved
2. Simple Strategy for Steering Polymer Film Formation on QCM Sensor Surfaces
Open this publication in new window or tab >>Simple Strategy for Steering Polymer Film Formation on QCM Sensor Surfaces
(English)Manuscript (preprint) (Other academic)
National Category
Analytical Chemistry
Research subject
Natural Science, Organic Chemistry
Identifiers
urn:nbn:se:lnu:diva-37250 (URN)
Available from: 2014-10-07 Created: 2014-09-24 Last updated: 2015-04-16Bibliographically approved
3. Biotin selective polymer nano-films
Open this publication in new window or tab >>Biotin selective polymer nano-films
Show others...
2014 (English)In: Journal of Nanobiotechnology, ISSN 1477-3155, E-ISSN 1477-3155, Vol. 12, 8Article in journal (Refereed) Published
Abstract [en]

Background: The interaction between biotin and avidin is utilized in a wide range of assay and diagnostic systems. A robust material capable of binding biotin should offer scope in the development of reusable assay materials and biosensor recognition elements. Results: Biotin-selective thin (3-5 nm) films have been fabricated on hexadecanethiol self assembled monolayer (SAM) coated Au/quartz resonators. The films were prepared based upon a molecular imprinting strategy where N, N'-methylenebisacrylamide and 2-acrylamido-2-methylpropanesulfonic acid were copolymerized and grafted to the SAM-coated surface in the presence of biotin methyl ester using photoinitiation with physisorbed benzophenone. The biotinyl moiety selectivity of the resonators efficiently differentiated biotinylated peptidic or carbohydrate structures from their native counterparts. Conclusions: Molecularly imprinted ultra thin films can be used for the selective recognition of biotinylated structures in a quartz crystal microbalance sensing platform. These films are stable for periods of at least a month. This strategy should prove of interest for use in other sensing and assay systems.

Keyword
Molecularly imprinted polymer, Biotinylated compounds, Photoinitiated graft co-polymerization, Quartz crystal microbalance (QCM) sensors, Sandwich-casting method
National Category
Organic Chemistry
Research subject
Chemistry, Organic Chemistry
Identifiers
urn:nbn:se:lnu:diva-34479 (URN)10.1186/1477-3155-12-8 (DOI)000335075000001 ()
Available from: 2014-05-28 Created: 2014-05-28 Last updated: 2017-02-16Bibliographically approved
4. Study of the Interaction of Trastuzumab and SKOV3 Epithelial Cancer Cells Using a Quartz Crystal Microbalance Sensor
Open this publication in new window or tab >>Study of the Interaction of Trastuzumab and SKOV3 Epithelial Cancer Cells Using a Quartz Crystal Microbalance Sensor
2015 (English)In: Sensors, ISSN 1424-8220, E-ISSN 1424-8220, Vol. 15, no 3, 5884-5894 p.Article 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
Keyword
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:nbn:se:lnu:diva-37513 (URN)10.3390/s150305884 (DOI)000354160900066 ()25763651 (PubMedID)
Available from: 2014-10-07 Created: 2014-10-07 Last updated: 2017-02-16Bibliographically approved

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