EBFP - Meercat

Clustergrammer Heatmap
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Figure 1C.zip2017-10-09112212
Figure 1D.zip2017-10-0912040401
Figure 4.zip2017-10-091224481304
Figure 5.zip2017-10-0911224241
Figure 6 Independent Expression.sky.zip2017-10-09112278
Figure 6 Coexpression.zip2017-10-09711392785585
MEERCAT: MULTIPLEXED EFFICIENT CELL FREE EXPRESSION OF RECOMBINANT QconCATS FOR LARGE SCALE ABSOLUTE PROTEOME QUANTIFICATION

  • Instrument: SCIEX QT5500
  • SpikeIn: Yes
Abstract
A major challenge in proteomics is the absolute accurate quantification of large numbers of proteins. QconCATs, artificial proteins that are concatenations of multiple standard peptides, are well established as an efficient means to generate standards for proteome quantification. Previously, QconCATs have been expressed in bacteria, but we now describe QconCAT expression in a robust, cell-free system. The new expression approach rescues QconCATs that previously were unable to be expressed in bacteria and can reduce the incidence of proteolytic damage to QconCATs. Moreover, it is possible to co-synthesise QconCATs in a highly-multiplexed translation reaction, co-expressing tens or hundreds of QconCATs simultaneously. By obviating bacterial culture and through the gain of high level multiplexing, it is now possible to generate tens of thousands of standard peptides in a matter of weeks, rendering absolute quantification of a complex proteome highly achievable in a reproducible, broadly deployable system.
Created on 10/9/17, 4:50 PM