Automated sample preparation method for mass spectrometry analysis on recombinant proteins.
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 4.5h and only small cultivation and buffer volumes are needed. In this proof of concept, 19,000-35,000Da recombinant proteins from both crude and clarified cell lysates were successfully prepared for subsequent analysis by electrospray ionization and matrix-assisted laser desorption/ionization mass spectrometry as well as by gel electrophoresis.
Steen
Johanna
J
School of Biotechnology, Department of Proteomics, AlbaNova University Center, Royal Institute of Technology, 106 91 Stockholm, Sweden.
Ramström
Margareta
M
Uhlén
Mathias
M
Hober
Sophia
S
Ottosson
Jenny
J
D001331:Automation; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*
D011994:Recombinant Proteins