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. 2019 Aug 8;14(8):e0221101. doi: 10.1371/journal.pone.0221101

Fig 3. MhPA14 purification and affinity for Superdex 200 size-exclusion resin.

Fig 3

A) SDS-PAGE tracking of MhPA14 purification through induction, extraction, and nickel-affinity chromatography. Lane 1 = pre-induction, Lane 2 = post-induction, Lane 3 = lysate insoluble fraction, Lane 4 = lysate soluble fraction, Lane 5 = elution from nickel-NTA agarose column. B) SDS-PAGE showing secondary purification of MhPA14 following nickel-affinity via anion-exchange chromatography (QSeph Lane), as well as the pooled fractions following S200-affinity elution with glucose and EDTA, respectively. C) Elution profiles of MhPA14 interacting with a Superdex 200 size-exclusion column under varying gradient conditions. The absolute absorbance at 280 nm (primary y-axis) was measured over 65 mL of buffer flow, during which an glucose gradient (top, red), an EDTA gradient (middle, green), or no gradient (bottom) was run.