title: Generation and optimization of human antagonistic antibodies against TIMP-1 as potential therapeutic agents in fibrotic diseases.
abstract: Impaired matrix metalloproteinase 1 (MMP-1) function, as result of the expression of increased levels of tissue inhibitor of metalloproteinase 1 (TIMP-1), plays an important role in the pathopysiolgical mechanism of fibrosis. In a recently performed clinically relevant rat animal model of established liver fibrosis, it could be shown, that blocking the interaction between the metalloproteinase and its inhibitor has beneficial effects in vivo. The rat TIMP-1 specific antagonistic antibody used in this study was derived from a human combinatorial antibody library (HuCAL) and blocks the interaction between rat TIMP-1 and MMP-13, the rat homologue of human MMP-1. We here describe the utilization of the same antibody source to generate fully human antibodies against human TIMP-1 which could be potential candidates for a therapy of fibrosis in man. In order to develop a highly potent antagonist of TIMP-1 action, antibodies isolated from the library were subjected to a number of different in vitro affinity maturation strategies. By these means, affinity and potency were improved by a factor of 87 and 65 fold, respectively, resulting in a valuable human therapeutic antibody candidate with a monovalent affinity of 150 pM and a potency for in vitro inhibition of TIMP-1/MMP-1 interaction of 200 pM.
authors.affiliation: MorphoSys AG, Lena-Christ-Str. 48, 82152 Martinsried/Planegg, Germany.
authors.forename: Bodo
authors.initials: B
authors.lastname: Brocks
authors.forename: Sabine
authors.initials: S
authors.lastname: Kraft
authors.forename: Stefan
authors.initials: S
authors.lastname: Zahn
authors.forename: Sabine
authors.initials: S
authors.lastname: Noll
authors.forename: Clark
authors.initials: C
authors.lastname: Pan
authors.forename: Michael
authors.initials: M
authors.lastname: Schauer
authors.forename: Barbara
authors.initials: B
authors.lastname: Krebs
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D000915:Antibody Affinity; D001709:Biotechnology / Q000379:methods; D001943:Breast Neoplasms / Q000628:therapy*; D002277:Carcinoma / Q000628:therapy; D020650:Combinatorial Chemistry Techniques; D022801:Complementarity Determining Regions; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000737:chemistry / Q000235:genetics / Q000276:immunology / Q000627:therapeutic use*; D008103:Liver Cirrhosis / Q000628:therapy*; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D019151:Peptide Library; D051381:Rats; D019715:Tissue Inhibitor of Metalloproteinase-1 / Q000276:immunology*
chemical_list: D000906:Antibodies; D022801:Complementarity Determining Regions; D007140:Immunoglobulin Fab Fragments; D019151:Peptide Library; D019715:Tissue Inhibitor of Metalloproteinase-1