Breakthroughs in Improving Antibody Affinity Maturation
Creative Biolabs is glad to release the unique antibody affinity maturation service for antibody
affinity maturation improvement.
As we know, antibodies are used extensively in diagnostics and as therapeutic agents. Achieving
high-affinity binding is crucial for expanding detection limits, extending dissociation half-times,
decreasing drug dosages and increasing drug efficacy. In order to conquer the shortages of
common antibodies, Creative Biolabs developed antibody affinity maturation service.
“We’ve gained extensive experience in antibody affinity maturation over the years. We use scFv as
the antibody format and monovalent display phagemid as the system to reduce the avidity effects
during antigen-binding screening”, said Dr. Monica Müller, chief scientific officer of Creative
Biolabs.
As Dr. Monica introduced, untargeted mutagenesis and oligonucleotide-directed mutagenesis are
utilized to construct random or defined sub-libraries to introduce a large number of mutants of the
original antibody. Antibody binders of higher affinity are then selected by increasing the screening
stringency. By constructing a series of sub-libraries of a scFv/Fab antibody, its protocol allows
increase of the affinity of the scFv antibodies from 10-8 to 10-9.
Untargeted Mutagenesis
It uses an error-prone PCR integrated DNA-shuffling approach to mutate main CDR regions
during sub-library construction. If the potential of introducing immunogenic mutations to
framework positions is not a concern, it usually uses this approach to create mutations at
completely random positions across the entire VH and VL fragments.
Oligonucleotide-directed Mutagenesis
If the structure of the antibody/antigen complex is available or modeling the structure of the
antibody/antigen is possible, certain positions can be randomized at a defined diversity to improve
the affinity. It is able to create any sub-libraries to incorporate the defined mutations utilizing
trimer codon technology.
Phage Display Antibody Library Screening
Subsequent library screening will fish out the antibody mutants that have high affinity. Two library
screening strategies are available, "surface-panning" strategy and "solution-sorting" strategy. In
the first "surface-panning" strategy, decreasing concentrations of antigen is surface immobilized.
In the second "solution-sorting" strategy, it has two approaches, selection based on the equilibrium
constant (Kd) and selection based on binding kinetics.
In addition to the three introduced above, Creative Biolabs has other approaches used, such as
antibody affinity measurement and peptide affinity maturation. Besides, single domain antibodies
affinity maturation and scFv affinity improvement are also available in Creative Biolabs.
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