title: Engineering and characterization of the chimeric antibody that targets the C-terminal telopeptide of the α2 chain of human collagen I: a next step in the quest to reduce localized fibrosis.
abstract: Inhibition of the extracellular process of collagen fibril formation represents a new approach to limiting posttraumatic or postsurgical localized fibrosis. It has been demonstrated that employing a monoclonal antibody that targets the C-terminal telopeptide of the α2 chain of collagen I blocks critical collagen I-collagen I interaction, thereby reducing the amount of collagen deposits in vitro and in animal models. Here, we developed a chimeric variant of a prototypic inhibitory antibody of mouse origin. The structure of this novel antibody was analyzed by biochemical and biophysical methods. Moreover, detailed biochemical and biological studies were employed to test its antigen-binding characteristics. The ability of the chimeric variant to block formation of collagen fibrils was tested in vitro and in high-density cultures representing fibrotic processes occurring in the skin, tendon, joint capsule, and gingiva. The potential toxicity of the novel chimeric antibody was analyzed through its impact on the viability and proliferation of various cells and by testing its tissue cross-reactivity in sets of arrays of human and mouse tissues. Results of the presented studies indicate that engineered antibody-based blocker of localized fibrosis is characterized by the following: (1) a correct IgG-like structure, (2) high affinity and high specificity for a defined epitope, (3) a great potential to limit the accumulation of collagen-rich deposits, and (4) a lack of cytotoxicity and nonspecific tissue reactivity. Together, the presented study shows the great potential of the novel chimeric antibody to limit localized fibrosis, thereby setting ground for critical preclinical tests in a relevant animal model.
authors.affiliation: Department of Orthopaedic Surgery, Jefferson Medical College, Thomas Jefferson University, Philadelphia, PA 19107, USA.
authors.forename: Jolanta
authors.initials: J
authors.lastname: Fertala
authors.forename: Andrzej
authors.initials: A
authors.lastname: Steplewski
authors.forename: James
authors.initials: J
authors.lastname: Kostas
authors.forename: Pedro
authors.initials: P
authors.lastname: Beredjiklian
authors.forename: Gerard
authors.initials: G
authors.lastname: Williams
authors.forename: William
authors.initials: W
authors.lastname: Arnold
authors.forename: Joseph
authors.initials: J
authors.lastname: Abboud
authors.forename: Anshul
authors.initials: A
authors.lastname: Bhardwaj
authors.forename: Cheryl
authors.initials: C
authors.lastname: Hou
authors.forename: Andrzej
authors.initials: A
authors.lastname: Fertala
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology*; D001483:Base Sequence; D015374:Biosensing Techniques; D016466:CHO Cells; D002470:Cell Survival; D024042:Collagen Type I / Q000276:immunology*; D006224:Cricetinae; D003412:Cricetulus; D024022:Fibrillar Collagens / Q000378:metabolism; D005355:Fibrosis / Q000276:immunology / Q000473:pathology; D006801:Humans; D007070:Immunoglobulin A / Q000378:metabolism; D007140:Immunoglobulin Fab Fragments / Q000378:metabolism; D007074:Immunoglobulin G / Q000378:metabolism; D007143:Immunoglobulin Heavy Chains / Q000378:metabolism; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D010455:Peptides / Q000276:immunology*; D016133:Polymerase Chain Reaction; D011485:Protein Binding; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000276:immunology*
chemical_list: D000906:Antibodies; D024042:Collagen Type I; D024022:Fibrillar Collagens; D007070:Immunoglobulin A; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D007143:Immunoglobulin Heavy Chains; D007147:Immunoglobulin Light Chains; D007135:Immunoglobulin Variable Region; D010455:Peptides; D011994:Recombinant Proteins; C052929:collagen type I trimeric cross-linked peptide