title: Generation of a potent recombinant homophilic chimeric anti-CD20 antibody.
abstract: Previously we increased the potency of therapeutic antibodies in targeting, induction of apoptosis, and growth inhibition in vitro and in vivo by chemically conjugating a homophilic peptide to the antibody. Here, we describe the construction of a chimeric fusion gene derived from the murine anti-CD20 antibody (1F5) variable region, with an engineered homophilic domain at the C-terminus of the human IgG1 sequence. The construct was expressed in CHO suspension cells and purified. The potency of the homophilic anti-CD20 antibody was compared to a chimeric antibody without the engineered homophilic domain. In this comparison, the homophilic anti-CD20 antibody showed increased binding to a human CD20 cell line, and significantly more ADCC, CDC, and induction of apoptosis in three cell lines. In addition, the homophilic anti-CD20 antibody demonstrated increased inhibition of proliferation of two cell lines. These data show that homophilic fusion protein antibodies with enhanced therapeutic potency can be produced with industry-standard fermentation protocols.
authors.affiliation: Department of Microbiology/Immunology, University of Kentucky, 600 Rose Street, Lexington, KY 40536-0096E, USA. hkohl00@uky.edu
authors.forename: Heinz
authors.initials: H
authors.lastname: Kohler
authors.forename: Kyle
authors.initials: K
authors.lastname: Rector
authors.forename: Jean
authors.initials: J
authors.lastname: Amick
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000494:pharmacology*; D000920:Antibody-Dependent Cell Cytotoxicity; D018951:Antigens, CD20 / Q000276:immunology*; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D003001:Cloning, Molecular; D004354:Drug Screening Assays, Antitumor; D006801:Humans; D006825:Hybridomas; D007963:Leukocytes, Mononuclear; D008223:Lymphoma; D051379:Mice; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000494:pharmacology*
chemical_list: D000906:Antibodies; D018951:Antigens, CD20; D000970:Antineoplastic Agents; D011993:Recombinant Fusion Proteins