Affinity maturation refers to the process of improving antibody affinity and binding interactions to target antigens. This is done naturally in vivo by somatic hypermutation and clonal selection in mammalian B cells, but it can also be done in the lab in vitro by mutagenesis and selection for therapeutic applications.
In vivo
When a host is repeatedly exposed to the same antigen, the antibodies produced by their immune response will have increasingly greater affinity, avidity, and anti-pathogen activity. This occurs due to somatic hypermutation in the variable CDRs (complementarity-determining regions) of immunoglobulin genes in B cells, and takes place in germinal centers (structures within secondary lymphoid tissues). Clonal selection takes place when follicular dendritic cells of the germinal centers present antigens to the B cells, with only the most competitive B cells surviving to stably conjugate with follicular B helper T cells. After several rounds of selection, high-affinity antibodies will be produced.1
In vitro
Similar to the in vivo process, in vitro affinity maturation is used to optimize antibodies, antibody fragments, or other peptides by diversifying the antibody base sequence and isolating higher-affinity binders.
FCMES-AM™ (Full Coverage Mammalian Expression System for Affinity Maturation) is Biointron's proprietary platform for affinity maturation. Each amino acid in the CDR region will be mutated to the other 17 amino acids (except Cysteine and Methionine) with equal ratios. The clones will be expressed in a mammalian system, and an ELISA binding assay will be conducted as an initial test. This will be followed by SPR (surface plasmon resonance) or FACS (fluorescence-activated cell sorting) for further affinity measurements.The process is as follows:
Site-directed saturated mutation
High-throughput mammalian cell expression
ELISA and sequencing to identify hot spots
Combinatorial mutation design and characterization
At Biointron, we are dedicated to accelerating antibody discovery, optimization, and production. Our advantages in affinity maturation include:
No-bias Site Saturation Mutagenesis in CDR Regions
Guaranteed at least 5-fold Affinity Improvement
Our team of experts can provide customized solutions that meet your specific research needs. Contact us to learn more about our services and how we can help accelerate your research and drug development projects.
Doria-Rose, N. A., & Joyce, M. G. (2015). Strategies to guide the antibody affinity maturation process. Current Opinion in Virology, 11, 137. https://doi.org/10.1016/j.coviro.2015.04.002
Antibody specificity refers to an antibody's ability to selectively bind to a unique epitope on a target antigen while avoiding interactions with unrelated antigens. This property arises from the highly specialized antigen-binding site located in the variable region of the antibody, which determines its unique binding characteristics.
Antibody affinity refers to the strength of the binding interaction between a single antigen epitope and the paratope (binding site) of an antibody. This interaction is a fundamental measure of how well an antibody recognizes its specific antigen target.
Recombinant antibodies are produced using genetic engineering techniques, unlike traditional antibody production, where the immune system generates antibodies without direct control over their sequence. By introducing genes encoding antibody fragments into host cells, such as bacteria or mammalian cells, recombinant antibodies can be expressed, purified, and deployed for applications including research, diagnostics, and therapeutics.
Recombinant antibody expression is a biotechnological process that involves engineering and producing antibodies outside their natural context using recombinant DNA technology.
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