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Orientation and capturing of antibody affinity ligands: Applications to surface plasmon resonance biochips
Linköping University, Department of Physics, Chemistry and Biology, Biotechnology. Linköping University, Faculty of Science & Engineering.
Linköping University, Department of Physics, Chemistry and Biology, Biotechnology. Linköping University, The Institute of Technology.
2011 (English)In: Sensors and actuators. B, Chemical, ISSN 0925-4005, E-ISSN 1873-3077, Vol. 158, no 1, 265-270 p.Article in journal (Refereed) Published
Abstract [en]

A surface plasmon resonance (SPR) sensor chip with immobilized protein G was used for simultaneously capturing, purifying and orienting antibody ligands. The ligands were further stabilized by chemical cross-linking. This procedure of designing the sensor chip improved efficient use of the ligands and could prolong the analytical use. less thanbrgreater than less thanbrgreater thanThe procedure was evaluated on standard dextran-coated sensor chips onto which commercial semi-purified antibodies towards human serum albumin and human troponin where captured and used for analysing their antigens. less thanbrgreater than less thanbrgreater thanThe procedure demonstrates a general design approach for presenting the biorecognition element on a biosensor surface which enhances sensitivity, stability and selectivity at the same time as an impure ligand is purified.

Place, publisher, year, edition, pages
Elsevier , 2011. Vol. 158, no 1, 265-270 p.
Keyword [en]
Biosensor, Affinity interaction, SPR, Biacore, Protein G, Sensor chip
National Category
Engineering and Technology
Identifiers
URN: urn:nbn:se:liu:diva-71770DOI: 10.1016/j.snb.2011.06.017ISI: 000295500200037OAI: oai:DiVA.org:liu-71770DiVA: diva2:453964
Note

Funding Agencies|European Commission|LSHB-CT-2007-037636|

Available from: 2011-11-04 Created: 2011-11-04 Last updated: 2017-12-08
In thesis
1. Exploring the SPR methodology for monitoring of critical attributes in toxicity testing and bioproduction
Open this publication in new window or tab >>Exploring the SPR methodology for monitoring of critical attributes in toxicity testing and bioproduction
2012 (English)Licentiate thesis, comprehensive summary (Other academic)
Abstract [en]

Analysis of biological components is central in bioprocess monitoring, process control, product quality control and cell based toxicity assaying. One of these themes that is pursued in this thesis is the use of biosensors for monitoring of molecular markers, exploiting the natural selectivity of biomolecules. Another is the use of glycoconjugates to monitor the activity of biomolecules in a flu vaccine process is studied and were the sensor is based on the concept of weak affinity giving fast response time for the sensor.

A third theme is monitoring of cell cultures used for toxicity testing different protein markers is of interest.

When developing biosensor surfaces for new antigens commercial preparations of antibodies are often used. In this work we have chosen to look at lactate dehydrogenase (LDH) and describe the preparation and characterisation of antibody used in biosensor surface development.

The design of a sensor surface is important for the characteristics of a sensor. By binding antibodies in an oriented manner to the surface a better control of the properties of the antibodies is achieved. The demonstrated method also has the advantage of in situ purification and provides a flexible platform for antibody evaluation and sensor development.

In one sentence this thesis explores the possibility of utilizing recognition elements of a biosensor surface. In particular, surface plasmon resonance (SPR) is used as the primary biosensing tool, however most findings in are relevant for other biosensors.

Moreover, the thesis approaches existing bioanalytical impediments, such as purity and accessibility of the recognition elements on the sensor surface and preparation strategies to achieve this.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2012. 19 p.
Series
Linköping Studies in Science and Technology. Thesis, ISSN 0280-7971 ; 1517
National Category
Natural Sciences
Identifiers
urn:nbn:se:liu:diva-73410 (URN)LIU-TEK-LIC-2012:2 (Local ID)978-91-7519-971-9 (ISBN)LIU-TEK-LIC-2012:2 (Archive number)LIU-TEK-LIC-2012:2 (OAI)
Presentation
2012-01-31, Panck, Fysikhuset, Campus Valla, Linköpings Universitet, Linköping, 09:15 (Swedish)
Opponent
Supervisors
Available from: 2012-01-03 Created: 2012-01-03 Last updated: 2012-01-11Bibliographically approved
2. Microfluidic biosensor systems for cardiotoxicity assaying
Open this publication in new window or tab >>Microfluidic biosensor systems for cardiotoxicity assaying
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Toxicity screening is an important part of pharmaceutical development and early detection of toxic side effects provide the opportunity for early redesign or termination of unfeasible projects. Today toxicity testing is relying on experiments on animals. Ethical concerns, high costs and problems with interspecies variability in animal experiments have introduced incentives for cell-based toxicity assays. The recent development of stem cell technology have raised the hope for toxicity testing with higher predictivity that can reduce the amount of animals sacrificed, increase the patient safety and reduce the costs in pharmaceutical development.

Cell development and behavior is to a large extent dependent on the microenvironment. Microfluidic techniques can be used to build small-sized structures that provide the opportunity to introduce a high degree of control of the cell culture environment with features in cell sizes. In this thesis is demonstrated two different methods for infusing cells into microfluidic cell culture devices using either cells clustered in cardiac bodies during differentiation or cells pre-seeded in microporous carriers prior to infusion.

Microfluidic cell culture devices are well suited for optical  evaluation. Demonstrated in this thesis is fluorescent staining in combination with confocal microscopy as well as automated imaging with evaluation of beating frequency of cardiomyocyte cell clusters can be used to assess toxicity of cells cultured in microfluidic devices.

Biosensors use biological recognition elements to measure the presence of a chemical substance, for example low concentrations of biomarkers secreted by cells in a toxicity assay. Especially capacitive biosensors have shown very low limit of detection. In addition, protein G is demonstrated as an affinity ligand to capture IgG antibodies used as recognition element in a biosensor application or used for antibody screening.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2015. 49 p.
Series
Linköping Studies in Science and Technology. Dissertations, ISSN 0345-7524 ; 1678
National Category
Biological Sciences Physical Sciences
Identifiers
urn:nbn:se:liu:diva-118295 (URN)978-91-7519-046-4 (ISBN)
Public defence
2015-06-12, Visionen, B-huset, Campus Valla, Linköping, 14:00 (Swedish)
Opponent
Supervisors
Available from: 2015-05-26 Created: 2015-05-26 Last updated: 2015-10-02Bibliographically approved

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