title: Generation of inhibitory monoclonal antibodies targeting matrix metalloproteinase-14 by motif grafting and CDR optimization.
abstract: Matrix metalloproteinase-14 (MMP-14) plays important roles in cancer metastasis, and the failures of broad-spectrum MMP compound inhibitors in clinical trials suggested selectivity is critical. By grafting an MMP-14 specific inhibition motif into complementarity determining region (CDR)-H3 of antibody scaffolds and optimizing other CDRs and the sequences that flank CDR-H3, we isolated a Fab 1F8 showing a binding affinity of 8.3 nM with >1000-fold enhancement on inhibition potency compared to the peptide inhibitor. Yeast surface display and fluorescence-activated cell sorting results indicated that 1F8 was highly selective to MMP-14 and competed with TIMP-2 on binding to the catalytic domain of MMP-14. Converting a low-affinity peptide inhibitor into a high potency antibody, the described methods can be used to develop other inhibitory antibodies of therapeutic significance.
authors.affiliation: Department of Chemical and Environmental Engineering, University of California, Riverside 900 University Ave, Riverside, CA 92521, USA.
authors.forename: Dong Hyun
authors.initials: DH
authors.lastname: Nam
authors.affiliation: Department of Chemical and Environmental Engineering, University of California, Riverside 900 University Ave, Riverside, CA 92521, USA.
authors.forename: Kuili
authors.initials: K
authors.lastname: Fang
authors.affiliation: Department of Chemical and Environmental Engineering, University of California, Riverside 900 University Ave, Riverside, CA 92521, USA.
authors.forename: Carlos
authors.initials: C
authors.lastname: Rodriguez
authors.affiliation: Department of Chemical and Environmental Engineering, University of California, Riverside 900 University Ave, Riverside, CA 92521, USA.
authors.forename: Tyler
authors.initials: T
authors.lastname: Lopez
authors.affiliation: Department of Chemical and Environmental Engineering, University of California, Riverside 900 University Ave, Riverside, CA 92521, USA xge@engr.ucr.edu.
authors.forename: Xin
authors.initials: X
authors.lastname: Ge
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000235:genetics / Q000276:immunology*; D020134:Catalytic Domain; D002460:Cell Line; D022801:Complementarity Determining Regions / Q000276:immunology*; D053511:Matrix Metalloproteinase 14 / Q000737:chemistry / Q000276:immunology*; D008958:Models, Molecular; D015202:Protein Engineering
chemical_list: D000911:Antibodies, Monoclonal; D022801:Complementarity Determining Regions; D053511:Matrix Metalloproteinase 14
keywords: CDR grafting; inhibitory antibody; matrix metalloproteinase; phage display; synthetic library