A gutless adenoviral vector expressing full-length anti-Her2 antibody.
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.
Jiang
Ming-Hong
MH
Laboratory of Viral and Gene Therapy, Eastern Hepatobiliary Surgical Hospital, Second Military Medical University, Shanghai, China.
Chen
Lin
L
Li
Lin-Fang
LF
Wu
Hong-Ping
HP
Jiang
Li-Hua
LH
Qian
Yan-Zhen
YZ
Fang
Guo-En
GE
Xue
Xu-Chao
XC
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
D061067:Antibodies, Monoclonal, Humanized; D004254:DNA Nucleotidyltransferases; D018719:Erb-b2 Receptor Tyrosine Kinases; D000068878:Trastuzumab; C045119:FLP recombinase