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Table 12 Mixed-mode chromatography (MMC) in proinsulin (PI) and insulin purification

From: Downstream processing of recombinant human insulin and its analogues production from E. coli inclusion bodies

Protein

Resin ligand functional group (interactions)

Media

Comments

References

Sulfonated PI

Multimodal weak cation exchanger (ionic interaction, hydrogen bonding, and hydrophobic interaction)

Capto MMC

Binding and elution buffers contain 8 M urea

pH step elution from 5.2 to 8

Yield: 96%; purity: 82%

Ability to bind sample without prior dilution

GE app. Notes 28-9966-22 AA (2012)

Insulin

Multimodal strong anion exchanger (ionic interaction, hydrogen bonding and hydrophobic interaction)

CaptoAdhere

Elute with a buffer of pH 3.4–3.6

Two wash steps at a pH higher than the elution pH to remove the majority of the impurities from enzymatic digest

Watson et al. (2018)