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Automated sample preparation method for mass spectrometry analysis on recombinant proteins
KTH, Proteomik.
KTH, Proteomik.
KTH, Proteomik.
KTH, Proteomik.
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2009 (English)In: Journal of Chromatography A, ISSN 0021-9673, E-ISSN 1873-3778, Vol. 1216, no 20, 4457-4464 p.Article in journal (Refereed) Published
Abstract [en]

A completely automated procedure for the purification and desalting of proteins with a polyhistidine purification tag prior to mass spectrometry analysis is presented. The system is ideal for rapid quality control and optimization studies and it provides researchers with a straightforward, reliable tool for studies of recombinant proteins. Forty-eight samples can be prepared within 4.5 h and only small cultivation and buffer volumes are needed. In this proof of concept, 19,000-35,000 Da recombinant proteins from both crude and clarified cell lysates were successfully prepared for subsequent analysis by electrospray ionization anti matrix-assisted laser desorption/ionization mass spectrometry as well as by gel electrophoresis.

Place, publisher, year, edition, pages
2009. Vol. 1216, no 20, 4457-4464 p.
Keyword [en]
Mass spectrometry, High-throughput, Automated sample preparation, Recombinant proteins, IMAC, RPLC, affinity-chromatography, purification
National Category
Natural Sciences
URN: urn:nbn:se:uu:diva-172141DOI: 10.1016/j.chroma.2009.03.041ISI: 000265956100021OAI: oai:DiVA.org:uu-172141DiVA: diva2:513427
QC 20100525Available from: 2010-08-05 Created: 2012-04-02 Last updated: 2012-04-02

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Steen, JohannaRamström, MargaretaUhlén, MathiasHober, SophiaOttosson, Jenny
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