Sometimes, antibodies from naïve antibody library would have tendency showing lower affinity to the antigen. Therefore, to get high-affinity and antigen specific antibody for therapeutic application, we optimize the antibody selection methods or affinity maturation technology for improving the affinity and stability. For antibody affinity maturation, we generally use several technologies such as random mutagenesis, site-directed mutagenesis, mutational hotspot targeting and LC shuffling.
1HC random mutagenesis
- Error-prone PCR with low-fidelity polymerization condition can induce low-level random point mutation.
- Mutations by error-prone PCR can improve the specificity and affinity of the antibody.
2Somatic hypermutation-mimicking in vitro mutagenesis
- This system is the best model for affinity maturation based on the fact that the DNA of antibody variable region can be mutated in the process of in vivo affinity maturation.
- It is the typical method for affinity improvement, and because heavy chain usually determines the antigen binding property, we fix the heavy chain and exchange the light chain variable region for high-affinity binding to the antigen.