Antibody Affinity Maturation Service

Antibody affinity maturation is the process by which the immune system generates antibodies of higher affinities during a response to antigen. The process involves the introduction of somatic mutations in antibody genes, which increase affinity for the antigen. There are several strategies for antibody affinity maturation, including random mutagenesis, targeted mutagenesis, chain shuffling, and in silico approaches. Random mutagenesis involves introducing randomized unbiased mutations using methods such as error-prone polymerase chain reaction (PCR) or DNA shuffling. Targeted mutagenesis involves introducing mutations in the complementarity-determining regions (CDRs) of immunoglobulin genes. Chain shuffling involves diversifying the sequence of the antibody, displaying the mutant gene repertoire on filamentous phage, and selecting higher-affinity binders on antigen. In silico approaches involve computational modeling of the antibody-antigen interaction to predict the effect of mutations on affinity. The increased antibody affinity can arise from any one or any combination of multiple diverse strategies and mechanisms, including improved shape complementarity at the interface with antigen, increased buried surface area upon complex formation, additional interfacial polar or hydrophobic interactions, and preorganization or rigidification of the antigen-binding site.

Fusion Biolabs has developed a pipeline for antibody affinity maturation, which is based on the principles of integrated mutation, such as structure-based site-directed mutagenesis, Off-Rate Biopanning and have successfully been used to optimize antibodies, antibody fragments or other peptide molecules, such as mimotope. Either saturation mutagenesis, random mutagenesis or focused mutagenesis inside the CDRs are introduced using V-fusion PCR cloning technologies. Chain shuffling is also used to increase the genetical diversity. Two or three rounds of mutation and selection (panning) usually results in antibody fragments with affinity in the low nanomolar range (5-10,000 fold increase).

Case Study 3: Antibody Affinity Maturation Services

Deliverables

1) Affinity enhanced candidates’ sequences and their plasmids (top 5-25 binders that meet the criteria of better binding affinity);
2) Project report

Ordering Information

Affinity Enhanced Antibody Affinity Maturation or Antibody Optimization Services

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