title: A gutless adenoviral vector expressing full-length anti-Her2 antibody.
abstract: 1. Therapeutic monoclonal antibodies are increasingly being used in clinical cancer treatment, but their complex technology and high cost limit their use. Helper-dependent (HD) adenoviruses are among the most efficient and safe gene therapy vectors capable of mediating long-term expression. 2. Using Gateway (Invitrogen, San Diego, CA, USA) cloning technology, we constructed an HD–trastuzumab (TAb) plasmid carrying the full-length anti-HER2 antibody gene. Using an efficient recombinase, namely in vitro-evolved Flippase-expressing recombinase, to excise the helper virus packaging signal in producer cells, we developed a scalable HD vector production method. Antibody expression of HD-TAb in vitro was detected by ELISA and western blot. 3. The full-length antibody gene delivery system allowed for continuous production of a full-length antibody at a high concentration. Bioactive antibody macromolecules were generated via gene transfer in vitro. 4. In conclusion, HD adenoviral vectors can stably express a full-length antibody for prolonged periods without the difficulties associated with sophisticated antibody manufacture techniques and at a much lower cost. As a promising tool for gene therapy, this novel system can shorten the duration and reduce the expense of antibody development.
authors.affiliation: Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgical Hospital, Second Military Medical University, Shanghai, China.
authors.forename: Ming-Hong
authors.initials: MH
authors.lastname: Jiang
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.forename: Lin-Fang
authors.initials: LF
authors.lastname: Li
authors.forename: Hong-Ping
authors.initials: HP
authors.lastname: Wu
authors.forename: Li-Hua
authors.initials: LH
authors.lastname: Jiang
authors.forename: Yan-Zhen
authors.initials: YZ
authors.lastname: Qian
authors.forename: Guo-En
authors.initials: GE
authors.lastname: Fang
authors.forename: Xu-Chao
authors.initials: XC
authors.lastname: Xue
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D061067:Antibodies, Monoclonal, Humanized / Q000096:biosynthesis* / Q000235:genetics; D010582:Bacteriophage lambda / Q000235:genetics / Q000378:metabolism; D001666:Binding Sites, Antibody; D015153:Blotting, Western; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D004254:DNA Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005455:Fluorescent Antibody Technique; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D057809:HEK293 Cells; D006801:Humans; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000276:immunology* / Q000378:metabolism; D013997:Time Factors; D014162:Transfection; D000068878:Trastuzumab
chemical_list: D061067:Antibodies, Monoclonal, Humanized; D004254:DNA Nucleotidyltransferases; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C045119:FLP recombinase