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Protein Microarrays...
Protein Microarrays on Carboxymethylated Dextran Hydrogels : Immobilization, Characterization and Application
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- Zhou, Ye (författare)
- Linköpings universitet,Sensorvetenskap och Molekylfysik,Tekniska högskolan
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- Andersson, Olof, 1978- (författare)
- Linköpings universitet,Sensorvetenskap och Molekylfysik,Tekniska högskolan
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- Lindberg, Peter (författare)
- Biacore AB, Rapsgatan 7, S-754 50, Uppsala, Sweden
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- Liedberg, Bo (författare)
- Linköpings universitet,Sensorvetenskap och Molekylfysik,Tekniska högskolan
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(creator_code:org_t)
- 2004-05-25
- 2004
- Engelska.
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Ingår i: Microchimica Acta. - : Springer Science and Business Media LLC. - 0026-3672 .- 1436-5073. ; 147:1-2, s. 21-30
- Relaterad länk:
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http://urn.kb.se/res...
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https://urn.kb.se/re...
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https://doi.org/10.1...
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Abstract
Ämnesord
Stäng
- Tetraoctadecylammonium bromide (TOAB, (CH3(CH2)17)4N+Br–) has been used to print temporary hydrophobic barriers on carboxymethylated dextran (CMD) hydrogels to create a generic platform for protein microarray applications. The primary reason for printing temporary hydrophobic barriers is to prevent cross-contamination and overflow during microdrop dispensing. Equally important is to eliminate the risk for non-specific binding to the barriers during analyte exposure. This has been accomplished by introducing a regeneration step that removes the barriers after ligand immobilization. The overall fabrication process was characterized by microscopic wetting, atomic force microscopy, imaging ellipsometry, fluorescence microscopy, surface plasmon microscopy and biospecific interaction analysis. A series of model proteins including transferrin, Protein A, anti-myoglobin and bovine serum albumin was spotted into the TOAB-defined areas under different experimental conditions, e.g. at increased humidity and reduced substrate temperature or with glycerol as an additive in the protein solution. Much emphasis was devoted to studies aiming at exploring the homogeneity and activity of the immobilized proteins. The printed barriers were removed after protein immobilization using tert-n-butyl alcohol (TBA). TBA was found to be a very efficient agent as compared to previously used salt regeneration solutions, and the regeneration time could be reduced from 30 to 10 minutes. Finally, the potential of using the well established CMD hydrogel chemistry as a platform for protein microarrays was exploited using surface plasmon microscopy.
Ämnesord
- MEDICIN OCH HÄLSOVETENSKAP -- Medicinska och farmaceutiska grundvetenskaper -- Andra medicinska och farmaceutiska grundvetenskaper (hsv//swe)
- MEDICAL AND HEALTH SCIENCES -- Basic Medicine -- Other Basic Medicine (hsv//eng)
Nyckelord
- Reversible hydrophobic barrier
- microcontact printing
- piezodispensing
- protein microarrays
- surface plasmon microscopy
- Chemistry
- Kemi
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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