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Interaction Studies in Complex Fluids with Optical Biosensors
Linköping University, Department of Physics, Chemistry and Biology, Applied Physics. Linköping University, The Institute of Technology.
2008 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

In this thesis interactions in complex fluids, such as serum and meat juice, were analysed with optical biosensor techniques.

Panels of lectins immobilised on gold surfaces were used for investigation of differences in protein glycosylation pattern in sera and meat juices between various species. The present panel was also used for investigation of global glycosylation changes of serum proteins in type 1 diabetes patients. Biorecognition was evaluated with null ellipsometry and scanning ellipsometry combined with multivariate data analysis techniques (MVDA). Principal component analysis (PCA) showed that the lectin panel enabled discrimination between sera from the different species as well as for the different meat juices. The results also indicate that there is a measurable global alteration in glycosylation pattern of serum proteins in type 1 diabetic patients compared to healthy subjects. Using an artificial neuronal net (ANN), it was also possible to correctly categorise unknown serum samples into their respective class or group. The analytical potential of combining information from lectin panels with multivariate data analysis was thereby demonstrated.

Also, a sensitive and specific method based on surface plasmon resonance (SPR) for detection of insulin autoantibodies (IAA) in serum samples from individuals at high risk of developing type 1 diabetes (T1D) has been developed. When measuring trace molecules, such as autoantibodies, in undiluted sera with label-free techniques like SPR, non-specific adsorption of matrix proteins to the sensor surface is often a problem, since it causes a signal that masks the analyte response. The developed method is an indirect competitive immunoassay designed to overcome these problems. Today, IAA is mainly measured in radio immunoassays (RIAs), which are time consuming and require radioactively labelled antigen. With our SPR-based immunoassay the overall assay time is reduced by a factor of >100 (from 4 days to 50 min), while sensitivity is maintained at a level comparable to that offered by RIA. Finally, the assay was used in a screening study of newly diagnosed type 1 diabetes patients and non-diabetic subjects.

Place, publisher, year, edition, pages
Linköping: Linköping University Electronic Press, 2008. , p. 70
Series
Linköping Studies in Science and Technology. Dissertations, ISSN 0345-7524 ; 1195
Keywords [en]
Type 1 diabetes, Insulin, Lectin panel, Ellipsometry, Meat juice, Serum proteins
National Category
Chemical Sciences
Identifiers
URN: urn:nbn:se:liu:diva-14694ISBN: 9789173938525 (print)OAI: oai:DiVA.org:liu-14694DiVA, id: diva2:24277
Public defence
2008-09-19, Planck, Fysikhuset, Campus Norrköping, Linköpings universitet, Linköping, 10:15 (English)
Opponent
Supervisors
Available from: 2008-09-26 Created: 2008-09-19 Last updated: 2022-11-08Bibliographically approved
List of papers
1. Investigation of sera from various species by using lectin affinity arrays and scanning ellipsometry
Open this publication in new window or tab >>Investigation of sera from various species by using lectin affinity arrays and scanning ellipsometry
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2005 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 530, no 2, p. 167-171Article in journal (Refereed) Published
Abstract [en]

Serum proteins of different species and of different human blood groups exhibit various protein glycosylation patterns. Sera from human, pig, sheep and guinea pig have been applied to a panel of eight different lectins immobilized on a gold wafer. The biorecognition has been evaluated with scanning ellipsometry and the two-dimensional matrices obtained have been treated with image analysis and MVDA for evaluation. The results showed a clear difference in protein binding pattern between the different species and thereby separation of the different sera could be made. Dendograms indicate that human and pig sera are the most related of the four different sera investigated.

Keywords
Serum proteins, lectins, scanning ellipsometry, image analysis, multivariate data analysis
National Category
Chemical Sciences
Identifiers
urn:nbn:se:liu:diva-12466 (URN)10.1016/j.aca.2004.09.022 (DOI)
Available from: 2008-09-06 Created: 2008-09-06 Last updated: 2017-12-13Bibliographically approved
2. Biosensor discrimination of meat juice from various animals using a lectin panel and ellipsometry
Open this publication in new window or tab >>Biosensor discrimination of meat juice from various animals using a lectin panel and ellipsometry
2005 (English)In: Analytica Chimica Acta, ISSN 0003-2670, E-ISSN 1873-4324, Vol. 547, no 2, p. 229-236Article in journal (Refereed) Published
Abstract [en]

In this work, simple microcontact printed gold-wafers were used to make a lectin panel for investigation and discrimination of different meat juices from fresh meat of cattle, chicken, pig, cod, turkey and lamb. Seven different lectins were thus attached to gold surfaces using the streptavidin–biotin method. Lectins recognize and bind specifically to carbohydrate structures present on different proteins. The biorecognition was evaluated with null ellipsometry and the data obtained was related to an internal standard of lactoferrin. The data was evaluated with multivariate data analysis techniques to identify possible discrimination or grouping of data. Scanning ellipsometry was used for visualization of the binding pattern of the lectins and the meat juice proteins. The two-dimensional images obtained could be used to visualize the protein distribution, furthermore, to exclude anomalies. The results showed that the different meat juices from the six different species: cattle, chicken, pig, cod, turkey and lamb could be discriminated from each other. The results showed to be more repetitive for the mammalian meat juices. Using a simple model based on an artificial neuronal net, it was also possible to classify meat juices from the mammals investigated.

Keywords
Lectin panel; Ellipsometry; Meat juice; Multivariate data analysis
National Category
Chemical Sciences
Identifiers
urn:nbn:se:liu:diva-12467 (URN)10.1016/j.aca.2005.05.054 (DOI)
Available from: 2008-09-06 Created: 2008-09-06 Last updated: 2017-12-13Bibliographically approved
3. Detection of global glycosylation changes of serum proteins in type 1 diabetes using a lectin panel and multivariate data analysis
Open this publication in new window or tab >>Detection of global glycosylation changes of serum proteins in type 1 diabetes using a lectin panel and multivariate data analysis
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2008 (English)In: Talanta: The International Journal of Pure and Applied Analytical Chemistry, ISSN 0039-9140, E-ISSN 1873-3573, Vol. 76, no 2, p. 333-337Article in journal (Refereed) Published
Abstract [en]

Global glycosylation changes of serum proteins in type 1 diabetic patients have in this paper been investigated based on the interaction of the saccharide moiety of serum proteins with different lectins. Lectins are proteins, which bind carbohydrates specifically and reversibly. Panels with lectins of various carbohydrate specificities were immobilized on gold surfaces. Sera from healthy individuals, newly diagnosed type 1 diabetes patients and type 1 diabetes patients having had the disease for 4–6 years, respectively, were applied to the lectin panel. The biorecognition was evaluated with null ellipsometry. Data obtained were related to an internal standard of lactoferrin. Multivariate data analysis (MVDA) techniques were used to analyze data.

Principal component analysis showed that the lectin panel enabled discrimination between sera from the three different above-mentioned groups. Using an artificial neuronal net (ANN), it was possible to correctly categorize unknown serum samples into one of the three groups.

 

Keywords
Lectin panel, Glycosylation changes, Type 1 diabetes, Ellipsometry, MVDA
National Category
Chemical Sciences
Identifiers
urn:nbn:se:liu:diva-12468 (URN)10.1016/j.talanta.2008.02.046 (DOI)
Available from: 2008-09-06 Created: 2008-09-06 Last updated: 2017-12-13Bibliographically approved
4. An indirect competitive immunoassay for insulin autoantibodies based on surface plasmon resonance
Open this publication in new window or tab >>An indirect competitive immunoassay for insulin autoantibodies based on surface plasmon resonance
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2008 (English)In: Biosensors and Bioelectronics, ISSN 0956-5663, Vol. 24, no 4, p. 876-881Article in journal (Refereed) Published
Abstract [en]

We have developed a sensitive and specific method based on surface plasmon resonance (SPR) for detection of insulin autoantibodies (IAA) in serum samples from individuals at high risk of developing type 1 diabetes (T1D). When measuring trace molecules in undiluted sera with label-free techniques like SPR, non-specific adsorption of matrix proteins to the sensor surface is often a problem, since it causes a signal that masks the analyte response. The developed method is an indirect competitive immunoassay designed to overcome these problems. Today, IAA is mainly measured in radio immunoassays (RIAs), which are time consuming and require radioactively labeled antigen. With our SPR-based immunoassay the overall assay time is reduced by a factor of >100 (4 days to 50 min), while sensitivity is maintained at a level comparable to that offered by RIA.

Keywords
SPR, Type 1 diabetes, Insulin autoantibodies, Indirect competitive immunoassay, RIA
National Category
Other Basic Medicine
Identifiers
urn:nbn:se:liu:diva-12469 (URN)10.1016/j.bios.2008.07.018 (DOI)
Note
The status of article IV on the day of defence was: Accepted.Available from: 2008-09-06 Created: 2008-09-06 Last updated: 2018-01-13Bibliographically approved
5. Determination of insulin autoantibodies using surface plasmon resonance: A screening study of newly diagnosed type 1 diabetes patients
Open this publication in new window or tab >>Determination of insulin autoantibodies using surface plasmon resonance: A screening study of newly diagnosed type 1 diabetes patients
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2008 (English)Manuscript (preprint) (Other academic)
Abstract [en]

We have investigated the screening potential of a surface plasmon resonance (SPR)-based indirectcompetitive immunoassay for quantification of insulin autoantibodies (IAA) in sera from childrennewly diagnosed with type 1 diabetes (T1D), using a radioimmunoassay (RIA) as reference technique.The two methods agreed well with respect to sample classification of 54 sera from newly diagnosedT1D children and 32 reference sera from non-diabetic children. Interestingly, five samples from newlydiagnosed T1D patients classified as IAA-negative according to RIA were IAA-positive with the SPRbasedassay, suggesting that the SPR-based assay might provide a higher sensitivity than the referenceRIA. However, 14 percent of the analyzed samples (five samples from non-diabetics and seven fromnewly diagnosed T1D patients) gave rise to anomalously high and easily distinguishable responses withthe SPR-based method, precluding IAA-quantification. A considerable part of the paper is devoted to adiscussion of possible causes of these anomalous responses. They were not due to temporary changesin the status of the patients, such as infections at the time of sampling, and also not related tocomplement activation. It is speculated whether a plausible explanation should instead be sought in theexistence of anti-idiotypic antibodies to IAA.

National Category
Chemical Sciences
Identifiers
urn:nbn:se:liu:diva-12470 (URN)
Available from: 2008-09-06 Created: 2008-09-06 Last updated: 2015-10-05Bibliographically approved

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