[Expression and tumoricidal activity of a human-mouse chimeric anti-DR5 antibody through a Furin/2A peptide].

Abstract

OBJECTIVE:

To express a full-length human-mouse chimeric anti-DR5 antibody from a single open reading frame with tumoricidal activity to various cancer cells.

METHODS:

The heavy and light chains of chimeric antibody were joined by the Furin and 2A (F/2A) self-cleavage peptide and cloned into a lentiviral vector of pWPXL. Then the HEK293 cells were infected with the constructed expression vector pWPXL-HF2AL. Western blot, enzyme-linked immunosorbent assay (ELISA) and MTS assay were used to detect the chimeric antibody expression, cleavage, binding affinity to the antigen and tumoricidal activity to various tumor cells.

RESULTS:

The recombinant chimeric antibody was successfully expressed from a single open reading frame in pWPXL-HF2AL construct. And it possessed a similar binding affinity to the parental murine counterpoint and strong tumoricidal activity to various cancer cells. For example, on the concentration of 3 µg/ml, it made the relative cells viability of HCT116, SMMC7721, A549 and U251 down to 20.6% ± 2.6%, 35.1% ± 2.7%, 76.1% ± 6.1% and 15.6% ± 2.0% respectively.

CONCLUSIONS:

The human-mouse chimeric anti-DR5 antibody of F/2A peptide is successfully expressed. Possessing a strong tumoricidal activity in various cancer cells, it may provide a novel strategy for cancer biotherapy.

• Authors: Meng Li, Fu-jia Lü, Juan Shi, Yan-xin Liu, De-xian Zheng
• Journal: Zhonghua yi xue za zhi (Zhonghua Yi Xue Za Zhi) Vol. 91 Issue 10 Pg. 707-10 (Mar 15 2011) ISSN: 0376-2491 [Print] China
• PMID: 21600181 (Publication Type: English Abstract, Journal Article)

Chemical References

• Antibodies
• Antineoplastic Agents
• Recombinant Fusion Proteins
• Furin

Topics

• Animals
• Antibodies (genetics, metabolism)
• Antineoplastic Agents (metabolism)
• Blotting, Western
• Enzyme-Linked Immunosorbent Assay
• Furin (metabolism)
• Genetic Vectors
• HEK293 Cells
• Humans
• Mice
• Recombinant Fusion Proteins (biosynthesis, metabolism)
• Transfection