Abstract
Abstract This review article describes our strategy for directed evolution of catalytic antibodies in phage-displayed antibody (Fab) libraries. To evolve catalytic antibodies toward higher catalytic activity, we have reconstructed an enzyme-evolutionary process in vitro. Thus, a phage-displayed combinatorial library from a hydrolytic antibody, 6D9, generated by the conventional in vivo method by immunization with transition-state analog (TSA) 5, was screened against a newly devised TSA 7 to optimize the differential affinity for the transition state relative to the ground state. This method provided evolved mutants which exhibited 20-fold higher activity than the parent antibody, 6D9. Structural analysis revealed an advantage of in vitro evolution over in vivo evolution: an induced catalytic residue in the evolved catalytic antibody arises from double mutations in one codon, which rarely occur in somatic hypermutation in the immune response.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Disclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.