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 used 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.
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.
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.
About Creative Biolabs
Creative Biolabs is a biotech service provider located in New York. It is specialized in providing custom biotechnology and pharmaceutical services that cover the full scope of biotechnology needs of early drug discovery and development for years, including affinity maturation of antibodies.
Company Name: Creative Biolabs
Contact Person: Bella Smith
Address:45-1 Ramsey Road
State: New York
Country: United States