Metabolomics

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Monitoring recombinant protein expression in bacteria by rapid evaporative ionisation mass spectrometry


ABSTRACT:

RATIONALE: There is increasing interest in methods of direct analysis mass spectrometry that bypass complex sample preparation steps.

METHODS: One of the most interesting new ionisation methods is rapid evaporative ionisation mass spectrometry (REIMS) in which samples are vapourised and the combustion products are subsequently ionised and analysed by mass spectrometry (Synpat G2-si). The only sample preparation required was the recovery of a cell pellet from a culture that could immediately be analysed.

RESULTS: In this paper, we demonstrate that REIMS can be used to monitor the expression of heterologous recombinant proteins in Escherichia coli. Clear segregation was achievable between bacteria harvesting plasmids that were strongly expressed when compared to other cultures in which the plasmid did not result in the expression of large amounts of recombinant product.

CONCLUSION: REIMS has considerable potential as a near-instantaneous monitoring tool for protein production in a biotechnology environment.

INSTRUMENT(S): Direct infusion MS - negative

SUBMITTER: Joscelyn Sarsby 

PROVIDER: MTBLS957 | MetaboLights | 2022-01-28

REPOSITORIES: MetaboLights

Dataset's files

Source:
Action DRS
MTBLS957 Other
FILES Other
a_MTBLS957_Neg_DI-MS_Effects_Of_Induction.txt Txt
a_MTBLS957_Neg_DI-MS_Expression_Timecourse.txt Txt
a_MTBLS957_Neg_DI-MS_Expressors_vs_NonExpressors.txt Txt
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Publications

Monitoring recombinant protein expression in bacteria by rapid evaporative ionisation mass spectrometry.

Sarsby Joscelyn J   McLean Lynn L   Harman Victoria M VM   Beynon Robert J RJ  

Rapid communications in mass spectrometry : RCM 20191123


<h4>Rationale</h4>There is increasing interest in methods of direct analysis mass spectrometry that bypass complex sample preparation steps.<h4>Methods</h4>One of the most interesting new ionisation methods is rapid evaporative ionisation mass spectrometry (REIMS) in which samples are vapourised and the combustion products are subsequently ionised and analysed by mass spectrometry (Synapt G2si). The only sample preparation required is the recovery of a cell pellet from a culture that can be anal  ...[more]

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