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. 2016 Jul 18;11(7):e0158923. doi: 10.1371/journal.pone.0158923

Fig 1. Validation of CHIKV nsP1 purity and identity.

Fig 1

Recombinant CHIKV nsP1 was purified with 6X his-mediated affinity chromatography followed by size exclusion chromatography. (A) Early attempts to purify CHIKV nsP1 resulted in highly aggregated protein that eluted exculsively in the void volume (eluted around 55 mL) during gel filtration, while protein purified after optimization of expression and purification conditions eluted around 72 mL. (B) Eluates resulting from size exclusion chromatography were concentrated and analyzed with SDS-PAGE gel electrophoresis and Coomassie staining. (C) This band was cut out, digested with trypsin and analysed further with orbitrap mass spectrometry analysis.