liu.seSök publikationer i DiVA
Ändra sökning
RefereraExporteraLänk till posten
Permanent länk

Direktlänk
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf
Gradient Hydrogel Matrix for Microarray and Biosensor Applications: An Imaging SPR Study
Linköpings universitet, Institutionen för fysik, kemi och biologi, Sensorvetenskap och Molekylfysik. Linköpings universitet, Tekniska högskolan.
Linköpings universitet, Institutionen för fysik, kemi och biologi, Sensorvetenskap och Molekylfysik. Linköpings universitet, Tekniska högskolan.
Linköpings universitet, Institutionen för fysik, kemi och biologi, Sensorvetenskap och Molekylfysik. Linköpings universitet, Tekniska högskolan.
Linköpings universitet, Institutionen för fysik, kemi och biologi, Sensorvetenskap och Molekylfysik. Linköpings universitet, Tekniska högskolan.
2009 (Engelska)Ingår i: Biomacromolecules, ISSN 1525-7797, E-ISSN 1526-4602, Vol. 10, nr 1, s. 142-148Artikel i tidskrift (Refereegranskat) Published
Abstract [en]

A biosensor matrix based on UV-initiated graft copolymerized poly(ethylene glycol) methacrylate and 2-hydroxyethyl methacrylate has been studied using imaging surface plasmon resonance (iSPR). By using a photo mask and a programmable shutter to vary the exposure time laterally, a gradient of matrix spots with physical thicknesses ranging from a few to tens of nanometers was generated. To maximize the dynamic range, imaging SPR was employed in wavelength interrogation mode. By finding the minimum in the reflectance spectra from each pixel of an image, SPR wavelength maps were constructed. The shift in SPR wavelength upon biospecific interaction was then measured both as a function of matrix thickness and composition. The performance of the matrix was evaluated in terms of immobilization of human serum albumin, biomolecular interaction with its antibody, and nonspecific binding of human fibrinogen. In addition, a low molecular weight interaction pair based on a synthetic polypeptide and calmodulin was also studied to explore the size selectivity of the hydrogel matrix. Our results show that the gradient matrix exhibits excellent properties for quick evaluation and screening of optimal hydrogel performance. The mixed hydrogel matrices display very low levels of nonspecific binding. It is also evident that the low molecular weight calmodulin is capable of freely diffusing and interacting throughout the entire hydrogel matrix, whereas the much larger albumin and its corresponding antibody, in particular, are partly/completely hindered from penetrating the interior of the matrix. This size-selectivity is attributed to a significant UV-initiated cross-linking or branching of the matrix during fabrication and/or protein mediated multipoint attachment during immobilization.

Ort, förlag, år, upplaga, sidor
2009. Vol. 10, nr 1, s. 142-148
Nationell ämneskategori
Naturvetenskap
Identifikatorer
URN: urn:nbn:se:liu:diva-16523DOI: 10.1021/bm801029bOAI: oai:DiVA.org:liu-16523DiVA, id: diva2:158133
Tillgänglig från: 2009-01-30 Skapad: 2009-01-30 Senast uppdaterad: 2017-12-14
Ingår i avhandling
1. Imaging surface plasmon resonance
Öppna denna publikation i ny flik eller fönster >>Imaging surface plasmon resonance
2008 (Engelska)Doktorsavhandling, sammanläggning (Övrigt vetenskapligt)
Abstract [en]

The central theme of this thesis is the use of imaging Surface Plasmon Resonance (iSPR) as a tool in the characterization of surfaces with laterally varying properties. Within the scope of this work, an instrument for iSPR analysis was designed and built. SPR is a very sensitive technique for monitoring changes in optical properties in the immediate vicinity of a sensor surface, which is very useful in biosensing and surface science research. We have employed SPR in the Kretschmann configuration, wherein surface plasmons are excited by means of an evanescent field arising from total internal reflection from the backside of the sensor surface. In iSPR, the signal is the reflectivity of TM-polarized light which is measured using an imaging detector, typically a CCD camera. Advantages of this technique include extreme surface sensitivity and, because detection is done from the backside, compatibility with complex samples. In addition, SPR is a non-labeling technique, and in imaging mode, a lateral resolution in the µm range can be attained.

The imaging SPR instrument could be operated in either wavelength interrogation mode or in intensity mode. In the former case, the objective is to find the SPR wave-length, λSPR, which is the wavelength at which the reflected intensity is at a minimum. In intensity mode, a snapshot of the intensity reflectance is taken at a fixed wavelength hand incidence angle.

In biosensor science, the use of an imaging technique offers a major advantage by enabling parallelization and thereby increasing throughput. We have, for example, used iSPR in biochemical interaction analysis to monitor immobilization and specific binding to protein and synthetic polypeptide micro arrays. The primary interest has been the study of soft matter surfaces that possess properties interesting in the field of biomimetics or for applications in biosensing. Specifically, the surfaces studied in this thesis include patterned self-assembled monolayers of thiolates on gold, a graft polymerized poly(ethylene glycol) (PEG) based hydrogel, a dextran hydrogel, and a polyelectrolyte charge gradient. Our results show that the PEG-based hydrogel is very well suited for use as a platform in protein immobilization in an array format, owing to the very low unspecific binding. In addition, well defined microarray templates were designed by patterning of hydrophobic barriers on dextran and monolayer surfaces. A polypeptide affinity microarray was further designed and immobilized on such a patterned monolayer substrate, in order to demonstrate the potential of analyte quantification with high sensitivity over a large dynamic range.

Furthermore, iSPR was combined with electrochemistry to enable laterally resolved studies of electrochemical surface reactions. Using this combination, the electrochemical properties of surfaces patterned with self assembled monolayers can be studied in parallel, with a spatial resolution in the µm regime. We have also employed electrochemistry and iSPR for the investigation of potential and current density gradients on bipolar electrodes.

The imaging SPR instrument could be operated in either wavelength interrogation mode or in intensity mode. In the former case, the objective is to find the SPR wave-length, λSPR, which is the wavelength at which the reflected intensity is at a minimum. In intensity mode, a snapshot of the intensity reflectance is taken at a fixed wavelength hand incidence angle.In biosensor science, the use of an imaging technique offers a major advantage by enabling parallelization and thereby increasing throughput. We have, for example, used iSPR in biochemical interaction analysis to monitor immobilization and specific binding to protein and synthetic polypeptide micro arrays. The primary interest has been the study of soft matter surfaces that possess properties interesting in the field of biomimetics or for applications in biosensing. Specifically, the surfaces studied in this thesis include patterned self-assembled monolayers of thiolates on gold, a graft polymerized poly(ethylene glycol) (PEG) based hydrogel, a dextran hydrogel, and a polyelectrolyte charge gradient. Our results show that the PEG-based hydrogel is very well suited for use as a platform in protein immobilization in an array format, owing to the very low unspecific binding. In addition, well defined microarray templates were designed by patterning of hydrophobic barriers on dextran and monolayer surfaces. A polypeptide affinity microarray was further designed and immobilized on such a patterned monolayer substrate, in order to demonstrate the potential of analyte quantification with high sensitivity over a large dynamic range.Furthermore, iSPR was combined with electrochemistry to enable laterally resolved studies of electrochemical surface reactions. Using this combination, the electrochemical properties of surfaces patterned with self assembled monolayers can be studied in parallel, with a spatial resolution in the µm regime. We have also employed electrochemistry and iSPR for the investigation of potential and current density gradients on bipolar electrodes.

Ort, förlag, år, upplaga, sidor
Linköping: Linköping University Electronic Press, 2008. s. 68
Serie
Linköping Studies in Science and Technology. Dissertations, ISSN 0345-7524 ; 1205
Nationell ämneskategori
Kemi
Identifikatorer
urn:nbn:se:liu:diva-14923 (URN)978-91-7393-820-4 (ISBN)
Disputation
2008-09-26, Planck, Fysikhuset, Campus Valla, Linköpings universitet, Linköping, 10:15 (Engelska)
Opponent
Handledare
Tillgänglig från: 2008-09-30 Skapad: 2008-09-30 Senast uppdaterad: 2017-01-11Bibliografiskt granskad

Open Access i DiVA

Fulltext saknas i DiVA

Övriga länkar

Förlagets fulltextLink to Ph.D. thesis

Personposter BETA

Andersson, OlofLarsson (Kaiser), AndréasEkblad, TobiasLiedberg, Bo

Sök vidare i DiVA

Av författaren/redaktören
Andersson, OlofLarsson (Kaiser), AndréasEkblad, TobiasLiedberg, Bo
Av organisationen
Sensorvetenskap och MolekylfysikTekniska högskolan
I samma tidskrift
Biomacromolecules
Naturvetenskap

Sök vidare utanför DiVA

GoogleGoogle Scholar

doi
urn-nbn

Altmetricpoäng

doi
urn-nbn
Totalt: 5473 träffar
RefereraExporteraLänk till posten
Permanent länk

Direktlänk
Referera
Referensformat
  • apa
  • harvard1
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • oxford
  • Annat format
Fler format
Språk
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Annat språk
Fler språk
Utmatningsformat
  • html
  • text
  • asciidoc
  • rtf