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Showing 1 results for Recombinant Herceptin
Nafiseh Kiarostami , Jafar Amani , Abbas Ali Imani Fooladi , Ali Mirhosseini ,
Volume 21, Issue 2 (7-2019)
Abstract
Background and Objective: Breast cancer is one of the most common types of cancer among women. HER-2 molecule as the receptor of tyrosine kinase from the family of epithermal growth factor is a major cause of cancer. The Herceptin protein molecule, which is an anti-HER-2 antibody, can play an important role in the diagnosis and treatment of breast cancer. This study was done to subcloning of Herceptin gene, expression in the prokaryotic system (E.coli) and produce Herceptin recombinant protein for use in the treatment of breast cancer.
Methods: In this descriptive – laboratory study, Herceptin gene from synthesized construct was isolated by enzyme digestion, and then subcloned to the expression vector pET28a. Subcloning of the gene was confirmed using PCR and enzyme digestion. After transferring the vector into E.coli BL21 DE3, expression of the recombinant Herceptin gene was induced by IPTG. The recombinant protein was evaluated by SDS-PAGE and purified with Ni-NTA column chromatography and finally verified by western blotting using anti-histidine antibody. The survival of cells adjacent to recombinant Hercaptin by MTT was investigated.
Results: Following the subcloning of the Herceptin gene, PCR and enzyme digestion, the 741 fragment of the Herceptin gene was confirmed. Confirmation of Herceptin's recombinant protein and its evaluation on SDS-PAGE gel about 27 kDa was done. The recombinant protein was also confirmed with anti-histidine tag. The purified protein adjacent to the SKBR3 cell line was able to block the growth of cancer cells.
Conclusion: Regarding the expersion of HER2 antigen on surface of breast cancer cells, Herceptin can act as antibody blocker and it arrests the growth of breast cancer cells.
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