alanine scanning mutagenesis protocol

Creative Biolabs has gained extensive experience in antibody affinity maturation. We usually take scFv as the antibody format in affinity maturation. Also, a monovalent display phagemid system is used to reduce the avidity effects during antigen-binding screening. We also provide affinity maturation services for single domain antibodies. Two methods, untargeted mutagenesis and oligonucleotide-directed mutagenesis, are employed 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, our proprietary protocol allows increase of the affinity of the scFv antibodies from 10 -9 to 10 -10. We have successfully obtained a scFv antibody that has an extremely high affinity of 10 -12, whose binding to the antigen is essentially irreversible.