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Exploring the SPR methodology for monitoring of critical attributes in toxicity testing and bioproduction
Linköping University, Department of Physics, Chemistry and Biology, Biotechnology. Linköping University, The Institute of Technology.
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: urn:nbn:se:liu:diva-73410Local ID: LIU-TEK-LIC-2012:2ISBN: 978-91-7519-971-9 (print)OAI: oai:DiVA.org:liu-73410DiVA: diva2:471904
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
List of papers
1. Monitoring of influenza virus hemagglutinin in process samples using weak affinity ligands and surface plasmon resonance
Open this publication in new window or tab >>Monitoring of influenza virus hemagglutinin in process samples using weak affinity ligands and surface plasmon resonance
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2008 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 623, 66-75 p.Article in journal (Refereed) Published
Abstract [en]

Surface plasmon resonance (SPR) was used to screen the interaction between a variety of affinity ligands and hemagglutinin (HA) from human influenza virus, with the aim of identifying low affinity ligands useful for the development of a rapid bioanalytical sensor. Three sialic acid-based structures and four lectins were evaluated as sensor ligands. The sialic acid-based ligands included a natural sialic acid-containing glycoprotein, human α1-acid glycoprotein (α1-AGP), and two synthetic 6′-sialyllactose-conjugates, with varying degree of substitution. The interaction of HA with the four lectin-based ligands, concanavalin A (Con A), wheat germ agglutinin (WGA), Maackia amurensis lectin (MAL), and Sambucus nigra agglutinin (SNA), showed a wide variation of affinity strengths. Affinity and kinetics data were estimated. Strong affinities were observed for Con A, WGA, α1-AGP, and a 6′-sialyllactose-conjugate with a high substitution degree, and low affinities were observed for MAL and a 6′-sialyllactose-conjugate with low substitution.

The main objective, to identify a low affinity ligand which could be used for on-line monitoring and product quantification, was met by a 6′-sialyllactose–ovalbumin conjugate that had 0.6 mol ligand per mol carrier protein. The apparent affinity of this ligand was estimated to be 1.5 ± 0.03 μM (KD) on the SPR surface. Vaccine process samples containing HA were analyzed in the range 10–100 μg HA mL−1 and correlated with single-radial immunodiffusion. The coefficient of variation on the same chip was between 0.010 and 0.091.

Place, publisher, year, edition, pages
Elsevier, 2008
Keyword
Influenza virus hemagglutinin; Affinity ligand; Surface plasmon resonance; Low affinity; Weak affinity
National Category
Natural Sciences
Identifiers
urn:nbn:se:liu:diva-44210 (URN)10.1016/j.aca.2008.06.005 (DOI)76040 (Local ID)76040 (Archive number)76040 (OAI)
Available from: 2009-10-10 Created: 2009-10-10 Last updated: 2017-12-13Bibliographically approved
2. Initial development towards in vitro toxicity assay for lactate dehydrogenase using surface plasmon resonance
Open this publication in new window or tab >>Initial development towards in vitro toxicity assay for lactate dehydrogenase using surface plasmon resonance
(English)Manuscript (preprint) (Other academic)
Abstract [en]

Work with initial development towards in vitro toxicity assay for lactate dehydrogenase (LDH) using surface plasmon resonance is described. LDH is one of the important versatile biomarkers in in vitro hepatotoxicity. In this report LDH is detected by surface plasmon resonance using antibodies directly immobilized to the sensor surface. The assay is further extended with protein G immobilized to the surface to capture the antibody ligand. This has the advantage of simultaneously and on site purify and orient antibody ligand.

Keyword
Lactate dehydrogenase, surface plasmon resonance, in vitro toxicity, biosensor, protein G
National Category
Natural Sciences
Identifiers
urn:nbn:se:liu:diva-73409 (URN)
Available from: 2012-01-03 Created: 2012-01-03 Last updated: 2012-01-03Bibliographically approved
3. Orientation and capturing of antibody affinity ligands: Applications to surface plasmon resonance biochips
Open this publication in new window or tab >>Orientation and capturing of antibody affinity ligands: Applications to surface plasmon resonance biochips
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
Keyword
Biosensor, Affinity interaction, SPR, Biacore, Protein G, Sensor chip
National Category
Engineering and Technology
Identifiers
urn:nbn:se:liu:diva-71770 (URN)10.1016/j.snb.2011.06.017 (DOI)000295500200037 ()
Note

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

Available from: 2011-11-04 Created: 2011-11-04 Last updated: 2017-12-08

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Bergström, Gunnar

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