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Multiplexed quantif...
Multiplexed quantification of proteins adsorbed to surface-modified and non-modified microdialysis membranes
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- Dahlin, Andreas P (författare)
- Uppsala universitet,Mikrosystemteknik
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- Hjort, Klas (författare)
- Uppsala universitet,Mikrosystemteknik
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- Hillered, Lars (författare)
- Uppsala universitet,Neurokirurgi
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- Sjödin, Marcus O.D. (författare)
- Uppsala universitet,Analytisk kemi
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- Bergquist, Jonas (författare)
- Uppsala universitet,Analytisk kemi
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- Wetterhall, Magnus (författare)
- Uppsala universitet,Analytisk kemi
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(creator_code:org_t)
- 2011-12-08
- 2012
- Engelska.
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Ingår i: Analytical and Bioanalytical Chemistry. - : Springer Science and Business Media LLC. - 1618-2642 .- 1618-2650. ; 402:6, s. 2057-2067
- Relaterad länk:
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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
- A simple and straightforward method for discovery and quantification of proteins adsorbed onto delicate and sensitive membrane surfaces is presented. The adsorbed proteins were enzymatically cleaved while still adsorbed onto the membranes using an on-surface enzymatic digestion (oSED). This was followed by isobaric tagging, nanoliquid chromatography, and tandem mass spectrometry. Protein adsorption on tri-block copolymer Poloxamer 407 surface-modified microdialysis (MD) membranes were compared with protein adsorption on unmodified MD membranes. Ventricular cerebrospinal fluid (vCSF) kept at 37 °C was used as sample matrix. In total, 19 proteins were quantified in two biological replicates. The surface-modified membranes adsorbed 33% less proteins than control membranes and the most abundant proteins were subunits of hemoglobin and clusterin. The adsorption of clusterin on the modified membranes was on average 36% compared to control membranes. The most common protein in vCSF, Albumin, was not identified adsorbed to the surface at all. It was also experimentally verified that oSED, in conjunction with tandem mass spectrometry can be used to quantify femtomole amounts of proteins adsorbed on limited and delicate surfaces, such as MD membranes. The method has great potential and can be used to study much more complex protein adsorption systems than previously reported.
Ämnesord
- NATURVETENSKAP -- Kemi (hsv//swe)
- NATURAL SCIENCES -- Chemical Sciences (hsv//eng)
Nyckelord
- TECHNOLOGY
- TEKNIKVETENSKAP
- Teknisk fysik med inriktning mot mikrosystemteknik
- Engineering Science with specialization in Microsystems Technology
Publikations- och innehållstyp
- ref (ämneskategori)
- art (ämneskategori)
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