Antibody Affinity Maturation

Affinity maturation is the process to improve antibody affinity for an antigen. In vivo,
natural affinity maturation by the immune system takes place by somatic
hypermutation and clonal selection. In vitro, in the laboratory affinity maturation, can
be
obtained
by
mutation
and
selection.
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.
Antibody Affinity Measurement
We offer Biacore Analysis services for binding kinetic analyses of antibodies. We
typically capture the antibody on the chip and run antigen on top of the captured
antibody. The antigen will be ran at 6 different concentrations for each antibody and
chi-square analysis will be performed on the binding constants we obtain from each
antigen concentration. The documentation package will include a real time on-rate
(Ka), off rate (Kd), an affinity constant (KD), chi square value and a graph of realtime binding kinetics. We would like to obtain ~50 uL of 1 mg/mL antigen and
antibody solutions. We will need ~100 ug of antigen and ~50ug for each antibody. We
would need MW information for the antigen as well. It may require special
considerations for antigens with repeated or multiple epitopes for affinity
determination. More about antibody maturation.