title: Engineering and characterization of a baculovirus-expressed mouse/human chimeric antibody against transferrin receptor.
abstract: Transferrin receptor (TfR) has been explored as a target for antibody-based therapy of cancer. In the previous study, we reported a murine anti-TfR monoclonal antibody (mAb) 7579 had good anti-tumor activities in vitro. In an attempt to reduce its immunogenicity and enhance its ability to recruit immune effector mechanism in vivo, we herein developed its chimera in the baculovirus/insect cell expression system based on the mating-assisted genetically integrated cloning (MAGIC) strategy. The chimeric light and heavy chains, containing human IgG1 constant regions, were correctly processed and assembled in insect cells, and then secreted into the mediums as heterodimeric H(2)L(2) immunoglobulins. Furthermore, analyses of antigen-binding assay and competitive binding assay indicated that the chimeric antibody possessed specificity and affinity similar to that of its parental murine antibody. Results of the antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC) assay verified that the chimeric antibody could efficiently mediate ADCC and CDC against TfR-overexpressing tumor cells. These results suggested that this baculovirus-expressed chimeric anti-TfR IgG1 might have the potential to be used for cancer immunotherapy. Meanwhile, the MAGIC strategy, facilitating the rapid generation of chimeric mAbs, could be one of the efficient strategies for antibody engineering.
authors.affiliation: Department of Immunology, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, People's Republic of China.
authors.forename: Xin
authors.initials: X
authors.lastname: Shen
authors.forename: Gui-Bin
authors.initials: GB
authors.lastname: Hu
authors.forename: Si-Jing
authors.initials: SJ
authors.lastname: Jiang
authors.forename: Feng-Rong
authors.initials: FR
authors.lastname: He
authors.forename: Wei
authors.initials: W
authors.lastname: Xing
authors.forename: Li
authors.initials: L
authors.lastname: Li
authors.forename: Juan
authors.initials: J
authors.lastname: Yang
authors.forename: Hui-Fen
authors.initials: HF
authors.lastname: Zhu
authors.forename: Ping
authors.initials: P
authors.lastname: Lei
authors.forename: Guan-Xin
authors.initials: GX
authors.lastname: Shen
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry* / Q000235:genetics / Q000276:immunology; D000920:Antibody-Dependent Cell Cytotoxicity; D016367:Baculoviridae / Q000235:genetics*; D001667:Binding, Competitive; D002460:Cell Line; D006801:Humans; D006825:Hybridomas; D051379:Mice; D016133:Polymerase Chain Reaction; D015202:Protein Engineering; D011990:Receptors, Transferrin / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry* / Q000235:genetics / Q000276:immunology
chemical_list: D000906:Antibodies; D011990:Receptors, Transferrin; D011993:Recombinant Fusion Proteins