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Table 1 Protein engineering strategies to reduce aggregation or derived effects during either production or administration, illustrated by representative examples.

From: Post-production protein stability: trouble beyond the cell factory

Protein engineering strategy Result Protein Reference
Improving protein folding during production    
Cys→Ser point mutations Reduced aggregation, enhanced proteolytic stability bFGFa [100]
Point mutations in an hydrophobic stretch Reduced aggregation 11 beta-HSD1 [101]
Directed evolution/point mutations Reduced aggregation Cytochrome P450sca-2 [102]
Fusion of SUMO tag Improved refolding Fgf15 [103]
Polycationic amino acid tag fusion Reduced aggregation Candida antarctica lipase B [104]
Fusion to polylysines or polyarginines Reduced aggregation BPTI-22 [105]
Fusion to MBP Reduced aggregation Ribonuclease inhibitor [106]
Fusion to GrpE Reduced aggregation hIL-3 [107]
Fusion to NusA Reduced aggregation, enhanced proteolytic stability E8R viral protein [108]
Improving protein folding, stability and performance during administration    
Single amino acid substitution Inhibited oligomer formation; enhanced bioavailability Insulin Aspart ® [109]
Single amino acid substitution Improved folding INF-β-1b (Betaferon ®) [110]
N-terminal peptide deletion Enhanced stability KGF [111]
Fusion with albumin Extended half-life Albinterferon α-2b [76]
Fusion with transferrin Enhanced gastrointestinal adsorption hGH [75]
Artificial consensus protein sequence Enhanced activity Interferon αcon-1 (Infergen ®) [112]
Fusion of a HIV Tat segment Enhanced solubility p53 [113]
Fusion of a HIV Tat protein and ODD Enhanced stability and activity in hypoxic tumor cells Casp-3 [114]
Fusion of a HIV Tat protein and sequence modification Cell penetration and selective activation in HIV-infected cells Casp-3 [115]
Ligand incorporation (mainly antibody fragments) Enhanced stability and bioavailbility IL-2 [71]
  1. a Abbreviations are: 11 beta-HSD1, 11 beta-Hydroxysteroid dehydrogenase type 1; aFGF, acidic fibroblast growth factor; bFGF: Fgf15, Fibroblast growth factor 15; BPTI-22, Bovine pancreatic trypsin inhibitor variant 22; Casp-3, caspase 3; HIV, human immunodeficiency virus; hFGF, Human basic fibroblast growth factor; BSA, bovine serum albumin; HAS, human serum albumin; hGH, human growth hormone; hIL-3, human interleukin-3; KGF, keratinocyte growth factor; IL, interleukin; MAGOH, Protein mago nashi homolog; MBP, maltose-binding protein; OOD, oxygen-dependent degradation domain of hypoxia-inducible factor-1alpha; rhDNase, recombinant human DNAse; SUMO, small ubiquitin-related modifier.