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Affinity and size exclusion chromatography was used to purify protein X S-Supernatant pH 7.5 F-Flow-through pH 7.5 W-Wash pH 6.5 L-Ladder 10kDa 15kDa 27kDa 35kDa 55kDa E1-Elution 1 pH 4.0 E2-Elution 2 pH 4.0 E3-Elution 3 pH 4.0 E4-Elution 4 pH 4.0 E5-Elution 5 pH 4.0 L S F E1 E2 E3 E4 E5 W S F W E1 E2 E3 E4 E5 L 10kDa 15kDa 27kDa 35kDa 55kDa Nickel-NTA 16kDa 16kDa 15kDa 27kDa 35kDa 55kDa F1 F2 F3 F4 F5 F6 F7 F1-Fraction 1 pH 7.5 F2-Fraction 2 pH 7.5 F3-Fraction 3 pH 7.5 F4-Fraction 4 pH 7.5 F5-Fraction 5 pH 7.5 16kDa 15kDa 27kDa 35kDa 55kDa F1 F2 F3 F4 F5 Size exclusion 16kDa

Protein Work

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AffinityandsizeexclusionchromatographywasusedtopurifyproteinX

S-Supernatant pH 7.5F-Flow-through pH 7.5W-Wash pH 6.5 L-Ladder

10kDa

15kDa

27kDa

35kDa

55kDa

E1-Elution 1 pH 4.0 E2-Elution 2 pH 4.0E3-Elution 3 pH 4.0E4-Elution 4 pH 4.0E5-Elution 5 pH 4.0

L S F E1 E2 E3 E4 E5W

S F W E1 E2 E3 E4 E5L

10kDa

15kDa

27kDa

35kDa

55kDa

Nickel-NTA

16kDa

16kDa

15kDa

27kDa

35kDa

55kDaF1F2F3F4F5F6F7

F1-Fraction 1 pH 7.5F2-Fraction 2 pH 7.5F3-Fraction 3 pH 7.5F4-Fraction 4 pH 7.5F5-Fraction 5 pH 7.5

16kDa

15kDa

27kDa

35kDa

55kDaF1F2F3F4F5

Sizeexclusion

16kDa

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