A MALDI-TOF MS based method for the fast screening of recombinant protein high-yield strains
In this study, we presented a MALDI-TOF MS based approach for the screening of recombinant protein high-yield strains. It relies on the MALDI MS analysis of the affinity tag containing peptide that released from the targe t recombinant proteins expressed in host strains. This approach was illustrated by quantifying the His-tag containing target peptide which was removed from the recombinant protein of Src Homology 2 superbinder by tobacco etch virus protease . After the specific enrichment and detection of target peptide with the theoretical molecular weight of 1895.09 Da, we could achieve the identification of strains with different expression levels of Src Homology 2 superbinder. Furthermore, the screening of strains at the absolute quantitative level was realized by adding the synthesized heavy labeled peptide as internal standards, and the quantitative results have a good linear correlation. In general, based on the virtue of high speed, accuracy and sensitivity of MALDI-TOF MS, this approach has the potential to achieve high throughput detection, and then provide powerful technical support for the screening of recombinant protein high-yield strains.
危险品化学品经营许可证（带存储）