The construction and expression of a novel chimeric anti-DR5 antibody.
Human tumor necrosis factor related apoptosis inducing ligand (TRAIL) could selectively induce apoptosis in a variety of transformed cells and is currently being developed as a cancer therapeutic drug. Unfortunately it has been reported that some forms of TRAIL could cause damage to normal liver cells. In order to avoid this side effect, many groups are now developing agonistic monoclonal antibody (MAb) against DR5. Previously, we developed a MAb (mDRA6) against DR5 with apoptosis inducing ability. The application of mouse origin antibody is limited in clinical use because it can induce human anti-mouse antibody (HAMA) responses that can cause allergic reaction and damage in the human body. Therefore, the variable region genes of this MAb were cloned and ligated into chimeric antibody expression vector (pCMV-VH and pCMV-VL), generating chimeric anti-human DR5 MAb (cmDRA6) expression vectors. The two plasmids were introduced into 293T cells with Lipofectamine 2000. The cell culture supernatant was collected and the expression level of cmDRA6 was detected with standard ELISA method. The chimeric mDRA6 could bind to its target antigen as demonstrated by the results of both ELISA and Western blot. This study of mDRA6 has laid the solid foundation for further application research in the future.
Kunpeng
Zhao
Z
Institute of Immunology, Medical College of Henan University, PR China.
Yugang
Wang
W
Jugao
Chen
C
Yan
Li
L
Beifen
Shen
S
Yuanfang
Ma
M
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D001483:Base Sequence; D015153:Blotting, Western; D002460:Cell Line; D003001:Cloning, Molecular; D017931:DNA Primers / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D005822:Genetic Vectors / Q000235:genetics / Q000276:immunology; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand / Q000276:immunology* / Q000378:metabolism; D014162:Transfection
D000911:Antibodies, Monoclonal; D017931:DNA Primers; D053220:Receptors, TNF-Related Apoptosis-Inducing Ligand