Rapid and quantitative analysis of recombinant protein expression using pyrolysis mass spectrometry and artificial neural networks: application to mammalian cytochrome b5 in Escherichia coli.
Recombinant Escherichia coli clones encoding between 0 and 6 copies of the mammalian cytochrome b5 gene were subjected to pyrolysis mass spectrometry (PyMS). To deconvolute the pyrolysis mass spectra so as to obtain quantitative information on the amount of cytochrome b5 produced fully-interconnected feedforward artificial neural networks (ANNs) were studied. It was found that the combination of PyMS and ANNs could be used to predict the amount of cytochrome b5 expressed in E. coli. PyMS is a novel, convenient and rapid method for the screening and analysis of microbial and other cultures producing recombinant proteins.
Goodacre
R
R
Institute of Biological Sciences, University of Wales, Aberystwyth, Dyfed, UK.
Karim
A
A
Kaderbhai
M A
MA
Kell
D B
DB
D000818:Animals; D003001:Cloning, Molecular; D015786:Cytochromes b5 / Q000096:biosynthesis* / Q000235:genetics; D004926:Escherichia coli; D008322:Mammals; D013058:Mass Spectrometry / Q000379:methods; D016571:Neural Networks, Computer; D011994:Recombinant Proteins / Q000096:biosynthesis*
D011994:Recombinant Proteins; D015786:Cytochromes b5