Preparation of a monoclonal antibody against the carcinoembryonic antigen, glypican‑3

  • Authors:
    • Yongdong Zhang
    • Dongri Qiu
    • Ronghua Li
    • Yawu Liu
    • Shuainan Shi
    • Yuliang Wang
  • View Affiliations

  • Published online on: March 13, 2019     https://doi.org/10.3892/mmr.2019.10019
  • Pages: 3889-3895
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

The carcinoembryonic antigen, glypican‑3 (GPC3), is a putative therapeutic target and diagnostic marker of hepatoma. In the present study, a monoclonal antibody (mAb) specifically against GPC3 was obtained via cloning the sequence of GPC3 via polymerase chain reaction and inserting it into a pET16b vector prior to transfection into Escherichia coli (E. coli) BL21. BALB/c mice were immunized with 20 µg purified antigen by intrasplenic embedding. Splenocytes and mouse myeloma cells SP2/0 were fused; then, the hybridoma cells were screened by an indirect ELISA. The properties of the mAb were examined by western blotting and immunofluorescence analysis against the purified protein. The results revealed that the prokaryotic expression vector of GPC3 had been successfully generated and GPC3 was stably expressed in E. coli BL21. A stable hybridoma cell line, 2F3, was generated in the present study, which produced mAbs against GPC3. The mAb 2F3 had a high antibody titer and the isotype was identified as IgG1/κ; 2F3 hybridomas had a median chromosome number of 98. Western blot and immunofluorescence analyses revealed that 2F3 specifically recognized recombinant and native GPC3. The 2F3 clone was proposed as a stable secretor of this mAb against GPC3. The results of present study indicated that the successful preparation of recombinant GPC3 protein and an anti‑human GPC3 mouse mAb may be provide a basis for developments in the diagnosis and treatment of liver cancer.
View Figures
View References

Related Articles

Journal Cover

May-2019
Volume 19 Issue 5

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
x
Spandidos Publications style
Zhang Y, Qiu D, Li R, Liu Y, Shi S and Wang Y: Preparation of a monoclonal antibody against the carcinoembryonic antigen, glypican‑3. Mol Med Rep 19: 3889-3895, 2019
APA
Zhang, Y., Qiu, D., Li, R., Liu, Y., Shi, S., & Wang, Y. (2019). Preparation of a monoclonal antibody against the carcinoembryonic antigen, glypican‑3. Molecular Medicine Reports, 19, 3889-3895. https://doi.org/10.3892/mmr.2019.10019
MLA
Zhang, Y., Qiu, D., Li, R., Liu, Y., Shi, S., Wang, Y."Preparation of a monoclonal antibody against the carcinoembryonic antigen, glypican‑3". Molecular Medicine Reports 19.5 (2019): 3889-3895.
Chicago
Zhang, Y., Qiu, D., Li, R., Liu, Y., Shi, S., Wang, Y."Preparation of a monoclonal antibody against the carcinoembryonic antigen, glypican‑3". Molecular Medicine Reports 19, no. 5 (2019): 3889-3895. https://doi.org/10.3892/mmr.2019.10019