Engineering the variable region of therapeutic IgG antibodies

Figures & data

Table 1 Summary of variable region engineering of therapeutic IgG antibodies

Engineering	Effect	Reference
Antigen binding properties
Affinity maturation
Random mutagenesis	Improving therapeutic efficacy*1	21
Targeted mutagenesis		11
Shuffling approaches	Decreasing minimum effective plasma antibody concentration*1	12
In silico approaches		28
Altering the specificity
Random mutagenesis	Reducing the cross reactivity to other antigens	38
Targeted mutagenesis	Broadening the specificity to related antigens	44
	Improving the non-human species cross-reactivity	46
	Binding to two different antigen by single binding site	50
Pharmacokinetics
Isoelectric point engineering
Lowering isoelectric point	Reducing the non-specific clearance	56
Engineering pH dependency
Rapid dissociation in acidic condition	Reducing the antigen-mediated clearance*2	58
	Binding to multiple antigen by single antibody	58
Pharmaceutical properties
Thermal stability improvement
Optimizing hydrophobic core and charge cluster residues	Retaining biological activity during storage	65, 66
Optimizing conserved residues	Minimizing aggregation not to increase immunogenicity	65, 66
Removing hydrophobic surface residues	Improving expression yield	65, 66
Optimizing VH/VL interface residues		65, 66
Solubility improvement
Reducing the surface hydrophobicity	Enabling high concentration formulation	77, 78
Introducing an N-linked carbohydrate		77, 78
Modifying the isoelectric point		78
Chemical stability improvement
Avoiding deamidation and isomerization	Retaining biological activity during storage	83, 87
Avoiding Met and Trp oxidation	Facilitating quality control in manufacturing	85
Heterogeneity improvement
Avoiding glycosylation	Facilitating quality control in manufacturing	94
Immunogenicity
Humanization
CDR grafting	Reducing immunogenicity	111
SDR grafting		116
Recombinant antibody libraries		121
Deimmunization
In silico prediction of T-cell epitopes	Reducing immunogenicity	127
In vitro analysis of T-cell epitopes		125
Introducing regulatory T-cell epitope		133

*1 This may not be always the case for the antibodies targeting solid tumors and agonistic antibodies.

*2 Antibodies targeting membrane-bound antigen with internalizing property.

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