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Monitoring the expression and purification of recombinant proteins by MALDI-TOF mass spectrometry

2001; Elsevier BV; Volume: 29; Issue: 1 Linguagem: Inglês

10.1016/s0141-0229(01)00352-0

1879-0909

Josep Villanueva, Françesc Canals, Enrique Querol, Francesc Avilés,

Molecular Biology Techniques and Applications

1Samples coming from biologic sources usually contain several contaminants that interfere seriously with Mass Spectrometry (MS) measurements. In this paper we report the application of MALDI-TOF MS to monitor recombinant protein expression and purification. The technique is based on the use of a C18 resin to clean and concentrate proteins in batch. The utility of this method is demonstrated for samples coming from different bacterial cultures expressing secreted and intracellular proteins ranging from 4 to 53 kDa. MALDI-TOF MS of peptide and proteins can be accomplished directly from complex bacterial cultures or from any purification step in a few minutes using the conventional stainless steel sample targets, allowing for a nearly instantaneous monitoring of the nature and integrity of recombinant expression products.