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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)